Note: Descriptions are shown in the official language in which they were submitted.
CA 02458872 2006-09-22
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HUMAN UBIQUITIN-CONJUGATING ENZYME
This is a divisional application of Canadian
Patent Application Serial No. 2,200,371 filed on March 19,
1997.
TECHNICAL FIELD
The present invention relates to a gene useful as
an indicator in the prophylaxis, diagnosis and treatment of
diseases in humans. More particularly, it relates to a
novel human gene analogous to rat, inouse, yeast, nematode
and known human genes, among others, and utilizable, after
cDNA analysis thereof, chromosome mapping of cDNA and
function analysis of cDNA, in gene diagnosis using said gene
and in developing a novel therapeutic method. It should be
understood that the expression "the invention" and the like
encompasses the subject matter of both the parent and the
divisional applications.
BACKGROUND ART
The genetic information of a liviing thing has been
accumulated as sequences (DNA) of four bases, namely A, C, G
and T, which exist in cell nuclei. Said genetic information
has been preserved for line preservation and ontogeny of
each individual living thing.
In the case of human being, the number of said bases
is said to be about 3 billion (3 x 10g) and supposedly there
are 50 to 100 thousand genes therein. Such genetic information
serves to maintain biological phenomena in that regulatory
proteins, structural proteins and enzymes are produced via
such route that mRNA is transcribed from a gene (DNA) and then
translated into a protein. Abnormalities in said route from
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gene to protein translation are considered to be
causative of abnormalities of life supporting systems,
for example in cell proliferation and differentiation,
hence causative of various diseases.
As a result of gene analyses so far made, a
number of genes which may be expected to serve as useful
materials in drug development, have been found, for
example genes for various receptors such as insulin
receptor and LDL receptor, genes involved in cell
proliferation and differentiation and genes for metabolic
enzymes such as proteases, ATPase and superoxide
dismutases.
However, analysis of human genes and studies of
the functions of the genes analyzed and of the relations
between the genes analyzed and various diseases have been
just begun and many points remain unknown. Further
analysis of novel genes, analysis of the functions
thereof, studies of the relations between the genes
analyzed and diseases, and studies for applying the genes
analyzed to gene diagnosis or for medicinal purposes, for
instance, are therefore desired in the relevant art.
If such a novel human gene as mentioned above
can be provided, it will be possible to analyze the level
of expression thereof in each cell and the structure and
function thereof and, through expression product analysis
CA 02458872 2004-03-23
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and other studies, it may become possible to reveal the
pathogenesis of a disease associated therewith, for
example a genopathy or cancer, or diagnose and treat said
disease, for instance. It is an object of the present
invention to provide such a novel human gene.
For attaining the above object, the present
inventors made intensive investigations and obtained the
findings mentioned below. Based thereon, the present
invention has now been completed.
DISCLOSURE OF INVENTION
Thus, the present inventors synthesized cDNAs
based on mRNAs extracted from various tissues, inclusive
of human fetal brain, adult blood vessels and placenta,
constructed libraries by inserting them into vectors,
allowing colonies of Escherichia coli transformed with
said libraries to form on agar medium, picked up colonies
at random and transferred to 96-well micro plates and
registered a large number of human gene-containing E.
coli clones.
Each clone thus registered was cultivated on a
small size, DNA was extracted and purified, the four
base-specifically terminating extension reactions were
carried out by the dideoxy chain terminator method using
the cDNA extracted as a template, and the base sequence
of the gene was determined over about 400 bases from the
CA 02458872 2004-03-23
4
5' terminus thereof using an automatic DNA sequencer. Based
on the thus-obtained base sequence information, a novel
family gene analogous to known genes of animal and plant
species such as bacteria, yeasts, nematodes, mice and humans
was searched for.
The method of the above-mentioned cDNA analysis is
detailedly described in the literature by Fujiwara, one of
the present inventors [Fujiwara, Tsutomu, Saibo Kogaku (Cell
Engineering), 14, 645-654 (1995)].
Among this group, there are novel receptors, DNA
binding domain-containing transcription regulating factors,
signal transmission system factors, metabolic enzymes and so
forth. Based on the homology of the novel gene of the present
invention as obtained by gene analysis to the genes analogous
thereto, the product of the gene, hence the function of the
protein, can approximately be estimated by analogy.
Furthermore, such functions as enzyme activity and binding
ability can be investigated by inserting the candidate gene
into an expression vector to give a recombinant.
The present invention provides a pharmaceutical
composition for treating prostate cancer, the pharmaceutical
composition comprising an antibody against human TMP-2
protein together with a pharmaceutically acceptable carrier.
In one embodiment, the antibody is conjugated to a
therapeutic moiety which can be a cytotoxic agent or a
radioisotope.
According to the present invention, there are
provided a novel human gene characterized by containing a
nucleotide sequence coding for an amino acid sequence
defined by SEQ ID NO:1, :4, :7, :10, :13, :16, :19, :22,
:25, :28, :31, :34, :37 or 40, a human gene characterized
s._ _. --...._._.-..__ _ ... .: õ ... .
02458872 2004-03-23
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by containing the nucleotide sequence defined by SEQ ID
NO:2, :5, :8, :11, :14, :17, :20, :23, :26, :29, :32,
:35, :38 or :41, respectively coding for the amino acid
sequence mentioned above, and a novel human gene
characterized by the nucleotide sequence defined by SEQ
ID NO:3, :6, :9, :12, :15, :18, :21, :24, :27, :30, :33,
:36, :39 or :42.
The symbols used herein for indicating amino
acids, peptides, nucleotides, nucleotide sequences and so
on are those recommended by IUPAC and IUB or in "Guide-
line for drafting specifications etc. including
nucleotide sequences or amino acid sequences" (edited by
the Japanese Patent Office), or those in conventional use
in the relevant field of art.
As specific examples of such gene of the
present invention,.there may be mentioned genes deducible
from the DNA sequences of the clones designated as "GEN-
501D08", "GEN-080G01", "GEN-025F07", "GEN-076C09", "GEN-
331G07", "GEN-163D09", "GEN-078D05TA13", "GEN-423A12",
"GEN-092E10", "GEN-428B12", "GEN-073E07", "GEN-093E05"
and "GEN-077A09" shown later herein in Examples 1 to 11.
The respective nucleotide sequences are as shown in the
sequence listing.
These clones have an open reading frame
comprising nucleotides (nucleic acid) respectively coding
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for the amino acids shown in the sequence listing. Their
molecular weights were calculated at the values shown
later herein in the respective examples. Hereinafter,
these human genes of the present invention are sometimes
referred to as the designation used in Examples 1 to 11.
In the following, the human gene of the present
invention is described in further detail.
As mentioned above, each human gene of the
present invention is analogous to rat, mouse, yeast,
nematode and known human genes, among others, and can be
uti.lized in human gene analysis based on the information
about the genes analogous thereto and in studying the
function of the gene analyzed and the relation between
the gene analyzed and a disease. It is possible to use
said gene in gene diagnosis of the disease associated
therewith and in exploitation studies of said gene for
medicinal purposes.
The gene of the present invention is
represented in terms of a single-stranded DNA sequence,
as shown under SEQ ID N0:2. It is to be noted, however,
that the present invention also includes a DNA sequence
complementary to such a single-stranded DNA sequence and
a component comprising both. The sequence of the gene of
the present invention as shown under SEQ ID NO:3n - 1
(where n is an integer of 1 to 14) is merely an example
.. ..
...CA 02458872 2004-03-23
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of the codon combination encoding the respective amino
acid residues. The gene of the present invention is not
limited thereto but can of course have a DNA sequence in
which the codons are arbitrarily selected and combined
for the respective amino acid residues. The codon
selection can be made in the conventional manner, for
example taking into consideration the codon utilization
frequencies in the host to be used [Nucl. Acids Res., 9,
43-74 (1981)].
The gene of the present invention further
includes DNA sequences coding for functional equivalents
derived from the amino acid sequence mentioned above by
partial amino acid or amino acid sequence substitution,
deletion or addition. These polypeptides may be produced
by spontaneous modification (mutation) or may be obtained
by posttranslational modification or by modifying the
natural gene (of the present invention) by a technique of
genetic engineering, for example by site-specific
mutagenesis [Methods in Enzymology, 154, p. 350, 367-382
(1987); ibid., 100, p. 468 (1983); Nucleic Acids
Research, 12, p. 9441 (1984); Zoku Seikagaku Jikken Koza
(Sequel to Experiments in Biochemistry) 1, "Idensi
Kenkyu-ho (Methods in Gene Research) II", edited by the
Japan Biochemical Society, p. 105 (1986)] or synthesizing
mutant DNAs by a chemical synthetic technique such as the
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phosphotriester method or phosphoamidite method [J. Am.
Chem. Soc., 89, p. 4801 (1967); ibid., 91, p. 3350
(1969); Science, 150, p. 178 (1968); Tetrahedron Lett.,
22, p. 1859 (1981); ibid., 24, p. 245 (1983)], or by
utilizing the techniques mentioned above in combination.
The protein encoded by the gene of the present
invention can be expressed readily and stably by
utilizing said gene, for example inserting it into a
vector for use with a microorganism and cultivating the
microorganism thus transformed.
The protein obtained by utilizing the gene of
the present invention can be used in specific antibody
production. In this case, the protein producible in
large quantities by the genetic engineering technique
mentioned above can be used as the component-to serve as
an antigen. The antibody obtained may be polyclonal or
monoclonal and can be advantageously used in the
purification, assay, discrimination or identification of
the corresponding protein.
The gene of the present invention can be
readily produced based on the sequence information
thereof disclosed herein by using general genetic
engineering techniques [cf. e.g. Molecular Cloning, 2nd
Ed., Cold Spring Harbor Laboratory Press (1989); Zoku
Seikagaku Jikken Koza, "Idenshi Kenkyu-ho I, II and III",
..
....... -- --- c : .CA 02458872 2004-03-23 .... ----------- - ._ __
-9-
edited by the Japan Biochemical Society (1986)].
This can be achieved, for example, by selecting
a desired clone from a human cDNA library (prepared in
the conventional manner from appropriate cells of origin
in which the gene is expressed) using a probe or antibody
specific to the gene of the present invention (e.g. Proc.
Natl. Acad. Sci. USA, 78, 6613 (1981); Science, 222, 778
(1983)].
The cells of origin to be used in the above
method are, for example, cells or tissues in which the
gene in question is expressed, or cultured cells derived
therefrom. Separation of total RNA, separation and
purification of mRNA, conversion to (synthesis of) cDNA,
cloning thereof and so on can be carried out by
conventional methods. cDNA libraries are also commer-
cially available and such cDNA libraries, for example
various cDNA libraries available from Clontech Lab. Inc.
can also be used in the above method.
Screening of the gene of the present invention
from these cDNA libraries can be carried out by the
conventional method mentioned above. These screening
methods include, for example, the method comprising
selecting a cDNA clone by immunological screening using
an antibody specific to the protein produced by the
corresponding cDNA, the technique of plaque or colony
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02458872 2004-03-23 ------ ---`-`---'--...1 _ .._::....,.. .
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hybridization using probes selectively binding to the.
desired DNA sequence, or a combination of these. As
regards the probe to be used here, a DNA sequence
chemically synthesized based on the information about the
DNA sequence of the present invention is generally used.
It is of course possible to use the gene of the present
invention or fragments thereof as the proble.
Furthermore, a sense primer and an antisense
primer designed based on the information about the
partial amino acid sequence of a natural extract isolated
and purified from cells or a tissue can be used as probes
for screening.
For obtaining the gene of the present
invention, the technique of DNA/RNA amplification by the
PCR method [Science, 230, 1350-1354 (1984)] can suitably
be employed. Particularly when the full-length cDNA can
hardly be obtained from the library, the RACE method
(rapid amplification of cDNA ends; Jikken Igaku
(Experimental Medicine), 12 (6), 35-38 (1994)], in
particular the 5'RACE method [Frohman, M. A., et al.,
Proc. Natl. Acad. Sci. USA, 85, 8998-9002 (1988)] is
preferably employed. The primers to be used in such PCR
method can be appropriately designed based on the
sequence information of the gene of the present invention
as disclosed herein and can be synthesized by a
CA 02458872 2004-03-23 .. . . . :.......... ... -- - --.. __ .. _. _. _.. ..:
. , :. ;.., - ..-....
-il-
conventional method.
The amplified DNA/RNA fragment can be isolated
and purified by a conventional method as mentioned above,
for example by gel electrophoresis.
The nucleotide sequence of the thus-obtained
gene of the present invention or any of various DNA
fragments can be determined by a conventional method, for
example the dideoxy method [Proc. Nati. Acad. Sci. USA,
74, 5463-5467 (1977)] or the Maxam-Gilbert method
[Methods in Enzymology, 65, 499 (1980)]. Such nucleotide
sequence determination can be readily performed using a
commercially available sequence kit as well.
When the gene of the present invention is used
and conventional techniques of recombinant DNA technology
[see e.g. Science, 224, p. 1431 (1984); Biochem. Biophys.
Res. Comm., 130, p. 692 (1985); Proc. Natl. Acad. Sci.
USA, 80, p. 5990 (1983) and the references cited above]
are followed, a recombinant protein can be obtained.
More detailedly, said protein can be produced by
constructing a recombinant DNA enabling the gene of the
present invention to be expressed in host cells,
introducing it into host cells for transformation thereof
and cultivating the resulting transformant.
In that case, the host cells may be eukaryotic
or prokaryotic. The eukaryotic cells include vertebrate
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-12-
cells, yeast cells and so on, and the vertebrate cells
include, but are not limited to, simian cells named COS
cells [Cell, 23, 175-182 (1981)], Chinese hamster ovary
cells and a dihydrofolate reductase-deficient cell line
derived therefrom [Proc. Natl. Acad. Sci. USA, 77, 4216-
4220 (1980)] and the like, which are frequently used.
As regards the expression vector to be used
with vertebrate cells, an expression vector having a
promoter located upstream of the gene to be expressed,
RNA splicing sites, a polyadenylation site and a
transcription termination sequence can be generally used.
This may further have an origin of replication as
necessary. As an example of said expression vector,
there may be mentioned pSV2dhfr [Mol. Cell. Biol., 1, 854
(1981)], which has the SV40 early promoter. As for the
eukaryotic microorganisms, yeasts are generally and
frequently used and, among them, yeasts of the genus
Saccharomyces can be used with advantage. As regards the
expression vector for use with said yeasts and other
eukaryotic microorganisms, pAM82 [Proc. Natl. Acad. Sci.
USA, 80, 1-5 (1983)], which has the acid phosphatase gene
promoter, for instance, can be used.
Furthermore, a prokaryotic gene fused vector
can be preferably used as the expression vector for the
gene of the present invention. As specific examples of
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....,,,......CA 02458872 2004-03-23 " --`------------`.:__.-..:~...~W:~....--'
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said vector, there may be mentioned pGEX-2TK and pGEX-4T-
2 which have a GST domain (derived from S. japonicum)
with a molecular weight of 26,000.
Escherichia coli and Bacillus subtilis are
generally and preferably used as prokaryotic hosts. When
these are used as hosts in the practice of the present
invention, an expression plasmid derived from a plasmid
vector capable of replicating in said host organisms and
provided in this vector with a promoter and the SD (Shine
and Dalgarno) sequence upstream of said gene for enabling
the expression of the gene of the present invention and
further provided with an initiation codon (e.g. ATG)
necessary for the initiation of protein synthesis is
preferably used. The Escherichia coli strain K12, among
others, is preferably used as the host Escherichia coli,
and pBR322 and modified vectors derived therefrom are
generally and preferably used as the vector, while
various known strains and vectors can also be used.
Examples of the promoter which can be used are the
tryptophan (trp) promoter, lpp promoter, lac promoter and
PL/PR promoter.
The thus-obtained desired recombinant DNA can
be introduced into host cells for transformation by using
various general methods. The transformant obtained can
be cultured by a conventional method and the culture
_ CA 02458872 2004-03-23 ...:..---=---.-_.. . .. ._;'.._
-14-
leads to expression and production of the desired protein
encoded by the gene of the present invention. The medium
to be used in said culture can suitably be selected from
among various media in conventional use according to the
host cells employed. The host cells can be cultured
under conditions suited for the growth thereof.
In the above manner, the desired recombinant
protein is expressed and produced and accumulated or
secreted within the transformant cells or extracellularly
or on the cell membrane.
The recombinant protein can be separated and
purified as desired by various separation procedures
utilizing the physical, chemical and other properties
thereof [cf. e.g. "Seikagaku (Biochemistry) Data Book
II", pages 1175-1259, lst Edition, lst Printing,
published June 23, 1980 by Tokyo Kagaku Dojin; Bio-
chemistry, 25 (25), 8274-8277 (1986); Eur. J. Biochem.,
163, 313-321 (1987)]. Specifically, said=procedures
include, among others, ordinary reconstitution treatment,
treatment with a protein precipitating agent (salting
out), centrifugation, osmotic shock treatment,
sonication, ultrafiltration, various liquid chromato-
graphy techniques such as molecular sieve chromatography
(gel filtration), adsorption chromatography, ion exchange
chromatography, affinity chromatography and high-
.:.. :: :...: .. . : .. . .. _. .. ... -....
CA 02458872 2004-03-23 ---'--- -15-
performance liquid chromatography (HPLC), dialysis and
combinations thereof. Among them, affinity chromato-
graphy utilizing a column with the desired protein bound
thereto is particularly preferred.
Furthermore, on the basis of the sequence
information about the gene of the present invention as
revealed by the present invention, for example by
utilizing part or the whole of said gene, it is possible
to detect the expression of the gene of the present
invention in various human tissues. This can be
performed by a conventional method, for example by RNA
amplification by RT-PCR (reverse transcribed-polymerase
chain reaction) [Kawasaki, E. S., et al., Amplification
of RNA, in PCR Protocol, A guide to methods and
applications, Academic Press, Inc., San Diego, 21-27
(1991)], or by northern blotting analysis [Molecular
Cloning, Cold Spring Harbor Laboratory (1989)], with good
results.
The primers to be used in employing the above-
mentioned PCR method are not limited to any particular
ones provided that they are specific to the gene of the
present invention and enable the gene of the present
invention alone to be specifically amplified. They can
be designed or selected apropriately based on the gene
information provided by the present invention. They can
CA 02458872 2006-09-22 .
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have a partial sequence comprising about 20 to 30
nucleotides according to the established practice.
Suitable examples are as shown in Examples 1 to 11.
Thus, the present invention also provides
primers and/or probes useful in specifically detecting
such novel gene.
By using the novel gene provided by the present
invention, it is possible to detect the expression of
said gene in various tissues, analyze the structure and
function thereof and, further, produce the human protein
encoded by said gene in the manner of genetic-
enginnering. These make it possible to analyze the
expression product, reveal the pathology of a disease
associated therewith, for example a genopathy or cancer,
and diagnose and treat the disease.
The following drawings are referred to in the
examples.
Fig. 1 shows the result obtained by testing the
P14 kinase activity of NPIK in Example 9. Fig. 2 shows
TM
the effect of Triton X-100 and adenosine on NPIK
activity.
EXAMPLES
The following examples illustrate the present
invention in further detail.
Example 1
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GDP dissociation stimulator gene
(1) Cloning and DNA sequencing of GDP dissociation
stimulator gene
mRNAs extracted from the tissues of human fetal
brain, adult blood vessels and placenta were purchased
from Clontech and used as starting materials.
cDNA was synthesized from each mRNA and
inserted into the vector ).ZAPII (Stratagene) to thereby
construct a cDNA library (Otsuka GEN Research Institute,
Otsuka Pharmaceutical Co., Ltd.)
Human gene-containing Escherichia coli colonies
were allowed to form on agar medium by the in vivo
excision technique [Short, J. M., et al., Nucleic Acids
Res., 16, 7583-7600 (1988)]. Colonies were picked up at
random and human gene-containing Escherichia coli clones
were registered on 96-well micro plates. The clones
registered were stored at -80 C.
Each of the clones registered was cultured
overnight in 1.5 ml of LB medium, and DNA was extracted
and purified using a model PI-100 automatic plasmid
extractor (Kurabo). Contaminant Escherichia coli RNA was
decomposed and removed by RNase treatment. The DNA was
dissolved to a final volume of 30 l. A 2- l portion was
used for roughly checking the DNA size and quantity using
a minigel, 7 pl was used for sequencing reactions and the
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-18-
remaining portion (21 l) was stored as plasmid DNA at
4 C.
This method, after slight changes in the
program, enables extraction of the cosmid, which is
useful also as a probe for FISH (fluorescence in situ
hybridization) shown later in the examples.
Then, the dideoxy terminator method of Sanger
et al. [Sanger, F., et al., Proc. Natl. Acad. Sci. USA,
74, 5463-5467 (1977)] using T3, T7 or a synthetic
oligonucleotide primer or the cycle suquence method
[Carothers, A. M., et al., Bio. Techniques, 7, 494-499
(1989)] comprising the dideoxy chain terminator method
plus PCR method was carried out. These are methods of
terminating the extension reaction specifically to the
four bases using a small amount of plasmid DNA (about 0.1
to 0. 5pg ) as a template.
The sequence primers used were FITC
(fluorescein isothiocyanate)-labeled ones. Generally,
about 25 cycles of reaction were performed using Taq
polymerase. The PCR products were separated on a
polyacrylamide urea gel and the fluorescence-labeled DNA
fragments were submitted to an automatic DNA sequencer
(ALFTM DNA Sequencer; Pharmacia) for determining the
sequence of about 400 bases from the 5' terminus side of
cDNA.
CA 02458872 2006-09-22
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Since the 3' nontranslational region is high in
heterogeneity for each gene and therefore suited for
discriminating individual genes from one another,
sequencing was performed on the 3' side as well depending
on the situation.
The vast sum of nucleotide sequence information
obtained from the DNA sequencer was transferred to a 64-
bit DEC 3400 computer for homology analysis by the
computer. In the homology analysis, a data base
TM
(GenBank, EMBL) was used for searching according to the
UWGCG FASTA program [Pearson, W. R. and Lipman, D. J.,
Proc. Natl. Acad. Sci. USA, 85, 2444-2448 (1988)].
As a result of arbitrary selection by the above
method and of cDNA sequence analysis, a clone designated
as GEN-501D08 and having a 0.8 kilobase insert was found
to show a high level of homology to the C terminal region
of the human Ral guanine nucleotide dissociation
stimulator (Ra1GDS) gene. Since Ra1GDS is considered to
play a certain role in signal transmission pathways, the
whole nucleotide sequence of the cDNA insert portion
providing the human homolog was further determined.
Low-molecular GTPases play an important role in
transmitting signals for a number of cell functions
including cell proliferation, differentiation and
transformation [Bourne, H. R. et al., Nature, 348, 125-
_ :....: .., .
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-20-
132 (1990); Bourne et al., Nature, 349, 117-127 (1991)].
It is well known that, among them, those
proteins encoded by the ras gene family function as
molecular switches or, in other words, the functions of
the ras gene family are regulated by different conditions
of binding proteins such as biologically inactive GDP-
binding proteins or active GDP-binding proteins, and that
these two conditions are induced by GTPase activating
proteins (GAPs) or GDS. The former enzymes induce GDP
10. binding by stimulating the hydrolysis of bound GTP and
the latter enzyme induces the regular GTP binding by
releasing bound GDP [Bogusuki, M. S. and McCormick, F.,
Nature, 366, 643-654 (1993)].
Ra1GDS was first discovered as a member of the
ras gene family lacking in transforming activity and as a
GDP dissociation stimulator specific to RAS (Chardin, P.
and Tavitian, A., EMBO J., 5, 2203-2208 (1986); Albright,
C. F., et al., EMBO J., 12, 339-347 (1993)].
In addition to Ral, Ra1GDS was found to
function, through interaction with these proteins, as an
effector molecule for N-ras, H-ras, K-ras and Rap
[Spaargaren, M. and Bischoff, J. R., Proc. Nati. Acad.
Sci. USA, 91, 12609-12613 (1994)].
The nucleotide sequence of the cDNA clone
designated as GEN-501D08 is shown under SEQ ID NO:3, the
......, ...,_ ._-.. __..-._... .. ,-CA 02458872 2004-03-23
-21-
nucleotide sequence of the coding region of said clone
under SEQ ID NO:2, and the amino acid sequence encoded by
said nucleotide sequence under SEQ ID NO:l.
This cDNA comprises 842 nucleotides, including
an open reading frame comprising 366 nucleotides and
coding for 122 amino acids. The translation initiation
codon was found to be located at the 28th nucleotide
residue.
Comparison between the Ra1GDS protein known
among conventional databases and the amino acid sequence
deduced from said cDNA revealed that the protein encoded
by this cDNA is homologous to the C terminal domain of
human RalGDS. The amino acid sequence encoded by this
novel gene was found to be 39.5% identical with the C
terminal domain of Ra1GDS which is thought to be
necessary for binding to ras.
Therefore, it is presumable, as mentioned
above, that this gene product might interact with the ras
family proteins or have influence on the ras-mediated
signal transduction pathways. However, this novel gene
is lacking in the region coding for the GDS activity
domain and the corresponding protein seems to be
different in function from the GDS protein. This gene
was named human Ra1GDS by the present inventors.
(2) Northern blot analysis
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The expression of the Ra1GDS protein mRNA in
normal human tissues was evaluated by Northern blotting
using, as a probe, the human cDNA clone labeled by the
random oligonucleotide priming method.
The Northern blot analysis was carried out with
a human MTN blot (Human Multiple Tissue Northern blot;
Clontech, Palo Alto, CA, USA) according to the manufac-,
turer's protocol.
Thus, the PCR amplification product from the
above GEN-501D08 clone was labeled with [32P]-dCTP
(random-primed DNA labeling kit, Boehringer-Mannheim) for
use as a probe.
For blotting, hybridization was performed
overnight at 42 C in a solution comprising 50$
formamide/5 x SSC/50 x Denhardt's.solution/0.1$ SDS
(containing 100 pg/ml denatured salmon sperm DNA). After
washing with two portions of 2 x SSC/0.01$ SDS at room
temperature, the membrane filter was further washed three
times with 0.1 x SSC/0.05* SDS at 50 C for 40 minutes.
An X-ray film (Kodak) was exposed to the filter at -70 C
for 18 hours.
As a result, it was revealed that a 900-bp
transcript had been expressed in all the human tissues
tested. In addition, a 3.2-kb transcript was observed
specifically in the heart and skeletal muscle. The
: :
:. CA 02458872 2004 03 23 .. . :. : _ ... ... . ... .. .. .
-23-
expression of these transcripts differing in size may be
due either to alternative splicing or to cross
hybridization with homologous genes.
(3) Cosmid clone and chromosome localization by FISH
FISH was performed by screening a library of
human chromosomes cloned in the cosmid vector pWE15
using, as a probe, the 0.8-kb insert of the cDNA clone
[Sambrook, J., et al., Molecular Cloning, 2nd Ed., pp.
3.1-3.58, Cold Spring Harbor Laboratory Press, Cold
Spring Harbor, New York (1989)].
FISH for chromosome assignment was carried out
by the method of Inazawa et al. which comprises G-banding
pattern comparison for confirmation [Inazawa, J., et al.,
Genomics, 17, 153-162 (1993)].
For use as a probe, the cosmid DNA (0.5 pg)
obtained from chromosome screening and corresponding to
GEN-501D08 was labeled with biotin-16-dUTP by nick
translation.
To eliminate the background noise due to
repetitive sequences, 0.5 l of sonicated human placenta
DNA (10 mg/mi) was added to 9.5 pl of the probe solution.
The mixture was denatured at 80 C for 5 minutes and
admixed with an equal volume of 4 x SSC containing 20t
dextransulfate. Then, a denatured slide was sown with
the hybridization mixture and, after covering with
CA 02458872 2004-03-23 :. :. , :.o_..__.......
-24-
paraffin, incubated in a wet chamber at 37 C for 16 to 18
hours. After washing with 50% formamide/2 x SSC at 37 C
for 15 minutes, the slide was washed with 2 x SSC for 15
minutes and further with 1 x SSC for 15 minutes.
The slide was then incubated in 4 x SSC supple-
mented with "1g Block Ace" (trademark; Dainippon Pharma-
ceutical) containing avidin-FITC (5 g/ml) at 37 C for 40
minutes. Then, the slide was washed with 4 x SSC for 10
minutes and with 4 x SSC containing 0.05% Triton X-100
for 10 minutes and immersed in an antifading PPD solution
[prepared by adjusting 100 mg of PPD (Wako Catalog No.
164-015321) and 10 ml of PBS(-) (pH 7.4) to pH 8.0 with
0.5 M Na2C03/0.5 M NaHCO3 (9:1, v/v) buffer (pH 9.0) and
adding glycerol to make a total volume of 100 ml]
containing 1% DABCO [1% DABCO (Sigma) in PBS(-):glycerol
1:9 (v:v)], followed by counter staining with DAPI (4,6-
diamino-2-phenylindole; Sigma).
With more than 100 tested cells in the
metaphase, a specific hybridization signal was observed
on the chromosome band at 6p2l.3, without any signal on
other chromosomes. It was thus confirmed that the RalGDS
gene is located on the chromosome 6p21.3.
By using the novel human RalGDS-associated gene
of the present invention as obtained in this example, the
expression of said gene in various tissues can be
................ .. . -..
CA 02458872 2004-03-23
-25-
detected and the human Ra1GDS protein can be produced in
the manner of genetic engineering. These are expected to
enable studies on the roles of the expression product
protein and ras-mediated signals in transduction pathways
as well as pathological investigations of diseases in
which these are involved, for example cancer, and the
diagnosis and treatment of such diseases. Furthermore,
it becomes possible to study the development and progress
of diseases involving the same chromosomal translocation
of the Ra1GDS protein gene of the present invention, for
example tonic spondylitis, atrial septal defect,
pigmentary retinopathy, aphasia and the like.
Example 2
Cytoskeleton-associated protein 2 gene (CKAP2 gene)
(1) Cytoskeleton-associated protein 2 gene cloning and
DNA sequencing
cDNA clones were arbitrarily chosen from a
human fetal brain cDNA library in the same manner as in
Example 1 were subjected to sequence analysis and, as a
result, a clone having a base sequence containing the
CAP-glycine domain of the human cytoskeleton-associated
protein (CAP) gene and highly homologous to several CAP
family genes was found and named GEN-080G01.
Meanwhile, the cytoskeleton occurs in the
cytoplasm and just inside the cell membrane of eukaryotic
..... .
CA 02458872 2004-03 23
-26-
cells and is a network structure comprising complicatedly
entangled filaments. Said cytoskeleton is constituted of
microtubules composed of tubulin, microfilaments composed
of actin, intermediate filaments composed of desmin and
vimentin, and so on. The cytoskeleton not only acts as
supportive cellular elements but also isokinetically
functions to induce morphological changes of cells by
polymerization and depolymerization in the fibrous
system. The cytoskeleton binds to intracellular
organelles, cell membrane receptors and ion channels and
thus plays an important role in intracellular movement
and locality maintenance thereof and, in addition, is
said to have functions in activity regulation and mutual
information transmission. Thus it supposedly occupies a
very important position in physiological activity
regulation of the whole cell. In particular, the
relation between canceration of cells and qualitative
changes of the cytoskeleton attracts attention since
cancer cells differ in morphology and recognition
response from normal cells.
The activity of this cytoskeleton is modulated
by a number of cytoskeleton-associated proteins (CAPs).
One group of CAPs is characterized by a glycine motif
highly conserved and supposedly contributing to associ-
ation with microtubules [CAP-GLY domain; Riehemann, K.
.. ..:_... _.. - --= ---. .______. _. _ - - -
; _ -
= ------ .CA 02458872 2004-03-23 . _ . '. .. -=-..... -:'r_.._ . .
-27-
and Song, C., Trends Biochem. Sci., 18, 82-83 (1993)].
Among the members of this group of CAPs, there
are CLIP-170, 150 kDa DAP (dynein-associated protein, or
dynactin), D. melanoyaster GLUED, S. cerevisiae BIKl,
restin [Bilbe, G., et al., EMBO J., 11, 2103-2113
(1992)]; Hilliker, C., et al., Cytogenet. Cell Genet.,
65, 172-176 (1994)] and C. elegans 13.5 kDa protein
[Wilson, R., et al., Nature, 368, 32-38 (1994)]. Except
for the last two proteins, direct or indirect evidences
have suggested that they could interact with
microtublues.
The above-mentioned CLIP-170 is essential for
the in vitro binding of endocytic vesicles to
microtubules and colocalizes with endocytic organelles
(Rickard, J. E. and Kreis, T. E., J. Biol. Chem., 18, 82-
83 (1990); Pierre, P., et al., Cell, 70, 887-900 (1992)].
The above-mentioned dynactin is one of the
factors constituting the cytoplasmic dynein motor, which
functions in retrograde vesicle transport [Schroer, T. A.
and Sheetz, M. P., J. Cell Biol., 115, 1309-1318 (1991)]
or probably in the movement of chromosomes during mitosis
[Pfarr, C. M., et al., Nature, 345, 263-265 (1990);
Steuer, E. R., et al., Nature, 345, 266-268 (1990);
Wordeman, L., et al., J. Cell Biol., 114, 285-294
(1991)].
_:___.: -__. ., , _,,._ . . .. ..... . . ..:..:_.._ . _.. .. . . . - .
CA 02458872 2004-03-23
-28-
GLUED, the Drosouhila homolog of mammalian
dynactin, is essential for the viability of almost all
cells and for the proper organization of some neurons
(Swaroop, A., et al., Proc. Nati. Acad. Sci. USA, 84,
6501-6505 (1987); Holzbaur, E. L. P., et al., Nature,
351, 579-583 (1991)].
BIK1 interacts with microtubules and plays an
important role in spindle formation during mitosis in
yeasts (Trueheart, J., et al., Mol. Cell. Biol., 7, 2316-
2326 (1987); Berlin, V., et al., J. Cell Biol., 111,
2573-2586 (1990)].
At present, these genes are classified under
the term CAP family (CAPs).
As a result of database searching, the above-
mentioned cDNA clone of 463-bp (excluding the poly-A
signal) showed significant homology in nucleotide
sequence with the restin and CLIP-170 encoding genes.
However, said clone was lacking in the 5' region as
compared with the restin gene and, therefore, the
technique of 5' RACE [Frohman, M. A., et al., Proc. Natl.
Acad. Sci. USA, 85, 8998-9002 (1988)] was used to isolate
this missing segment.
(2) 5' RACE (5' rapid amplification of cDNA ends)
A cDNA clone containing the 5' portion of the
gene of the present invention was isolated for analysis
CA 02458872 2006-09-22
-29-
by the 5' RACE technique using a commercial kit (5'-Rapid
TM
AmpliFinder RACE kit, Clontech) according to the
manufacturer's protocol with minor modifications, as
follows.
The gene-specific primer P1 and primer P2 used
here were synthesized by the conventional method and
their nucleotide sequences are as shown'below in Table 1.
The anchor primer used was the one attached to the
commercial kit.
Table 1
Primer Nucleotide sequence
Primer PI 5'-ACACCAATCCAGTAGCCAGGCTTG-3'
Primer P2 5'-CACTCGAGAATCTGTGAGACCTACATACATGACG-3'
cDNA was obtained by reverse transcription of
0.1 pg of human fetal brain poly(A)+RNA by the random
hexamer technique using reverse transcriptase
(SuperscriptTM II, Life Technologies) and the cDNA was
amplified by the first PCR using the Plprimer and anchor
primer according to Watanabe et al. [Watanabe, T., et
al., Cell Genet., in press).
Thus, to 0.1 pg of the above-mentioned cDNA
were added 2.5 mM dNTP/1 x Taq buffer (Takara Shuzo)/0.2
TM
pM P1 primer, 0.2 pM adaptor primer/0.25 unit ExTaq
enzyme (Takara Shuzo) to make a total volume of 50 }il,
followed by addition of the anchor primer. The mixture
-. , .... . - ..
: .: CA 02458872 2004-03-23 :...- . - - - - ------, .
-30-
was subjected to PCR. Thus, 35 cycles of amplification
were performed under the conditions: 94 C for 45 seconds,
60 C for 45 seconds, and 72 C for 2 minutes. Finally,
the mixture was heated at 72 C for 5 minutes.
Then, 1 l of the 50-Nl first PCR product was
subjected to amplification by the second PCR using the
specific nested P2 primer and anchor primer.. The second
PCR product was analyzed by 1.5$ agarose gel
electrophoresis.
Upon agarose gel electrophoresis, a single
band, about 650 nucleotides in size, was detected. The
product from this band was inserted into a vector
(pT7Blue(R)T-Vector, Novagen) and a plurality of clones
with an insert having an appropriate size were selected.
Six of the 5' RACE clones obtained from the PCR
product had the same sequence but had different lengths.
By sequencing two overlapping cDNA clones, GEN-080G01 and
GEN-080G0149, the protein-encoding sequence and 5' and 3'
flanking sequences, 1015 nucleotides in total length,
were determined. Said gene was named cytoskeleton-
associated protein 2 gene (CKAP2 gene).
The nucleotide sequence obtained from the
above-mentioned two overlapping cDNA clones GEN-080G01
and GEN-080G0149 is shown under SEQ ID N0:6, the
nucleotide sequence of the coding region of said clone
. .,..,., .. . . .. . -... .. _ .
CA 02458872 2004-03-23 - - - - -. ---.. - - -..
-31-
under SEQ ID NO:5, and the amino acid sequence encoded by
said nucleotide sequence under SEQ ID N0:4.
As shown under SEQ ID NO:6, the CKAP2 gene had
a relatively.GC-rich 5' noncoding region, with incomplete
triplet repeats, (CAG)4(CGG)4(CTG)(CGG), occurring at
nucleotides 40-69.
ATG located at nucleotides 274-276 is the
presumable start codon. A stop codon (TGA) was situated
at nucleotides 853-855. A polyadenylation signal
(ATTAAA) was followed by 16 nucleotides before the
poly(A) start. The estimated open reading frame
comprises 579 nucleotides coding for 193 amino acid
residues with a calculated molecular weight of 21,800
daltons.
The coding region was further amplified by RT-
PCR, to eliminate the possibility of the synthetic
sequence obtained being a cDNA chimera.
(2) Similarity of CKAP2 to other CAPs
While sequencing of CKAP2 revealed homology
with the sequences of restin and CLIP-170, the homologous
region was limited to a short sequence corresponding to
the CAP-GLY domain. On the amino acid level, the deduced
CKAP2 was highly homologous to five other CAPs in this
domain.
CKAP2 was lacking in such other motif
....... . . ,_.:;. _ . -
----
CA 02458872 2004-03-23
~
-32-
characteristics of some CAPs as the alpha helical rod and
zinc finger motif. The alpha helical rod is thought to
contribute to dimerization and to increase the micro-
tubule binding capacity [Pierre, P., et al., Cell, 70,
887-900 (1992)]. The lack of the alpha helical domain
might mean that CKAP2 be incapable of homo or hetero
dimer formation.
Paralleling of the CAP-GLY domains of these
proteins revealed that other conserved residues other
than glycine residues are also found in CKAP2. CAPs
having a CAP-GLY domain are thought to be associated with
the activities of cellular organelles and the
interactions thereof with microtubules. Since it
contains a CAP-GLY domain, as mentioned above, CKAP2 is
placed in the family of CAPs.
Studies with mutants of Glued have revealed
that the Glued product plays an important role in almost
all cells [Swaroop, A., et al., Proc. Natl. Acad. Sci.
USA, 84, 6501-6505 (1987)] and that it has other neuron-
specific functions in neuronal cells [Meyerowitz, E. M.
and Kankel, D. R., Dev. Biol., 62, 112-142 (1978)].
These microtubule-associated proteins are thought to
function in vesicle transport and mitosis. Because of
the importance of the vesicle transport system in
neuronal cells, defects in these components might lead to
,.:=;~_r_:__.. ._ ._. -_... _
w. _.-_ -. . . --- -. :.
,_ -_-_-----
' ---- CA 02458872 2004-03-23
-33-
aberrant neuronal systems.
In view of the above, CKAP2 might be involved
in specific neuronal functions as well as in fundamental
cellular functions.
(3) Northern blot analysis
The expression of human CKAP2 mRNA in normal
human tissues was examined by Northern blotting.in the
same manner as in Example 1 (2) using the GEN-080G01
clone (corresponding to nucleotides 553-1015) as a probe.
As a result, in all the eight tissues tested,
namely human heart, brain, placenta, lung, liver,
skeletal muscle, kidney and pancreas, a 1.0 kb transcript
agreeing in size with the CKAP2 cDNA was detected. Said
1.0 kb transcript was expressed at significantly higher
levels in heart and brain than in the other tissues
examined. Two weak bands, 3.4 kb and 4.6 kb, were also
detected in all the tissues examined.
According to the Northern blot analysis, the
3.4 kb and 4.6 kb transcripts might possibly be derived
from the same gene coding for the 1.0 kb CKAP2 by
alternative splicing or transcribed from other related
genes. These characteristics of the transcripts may
indicate that CKAP2 might also code for a protein having
a CAP-GLY domain as well as an alpha helix.
(4) Cosmid cloning and chromosomal localization by
;..._ ---------- - -----
......... .. .:::._~. ,.... _ =
CA 02458872 2004-03-23
-34-
direct R-banding FISH
Two cosmids corresponding to the CKAP2 cDNA
were obtained. These two cosmid clones were subjected to
direct R-banding FISH in the same manner as in Example 1
(3) for chromosomal locus mapping of CKAP2.
For suppressing the background due to
repetitive sequences, a 20-fold excessive amount of human
Cot-I DNA (BRL) was added as described by Lichter et al.
[Lichter, P., et al., Proc. Natl. Acad. Sci. USA, 87,
6634-6638 (1990)]. A Provia 100 film (Fuji ISO 100; Fuji
Photo Film) was used for photomicrography.
As a result, CKAP2 was mapped on chromosome
bands 19q13.11-q13.12.
Two autosomal dominant neurological diseases
have been localized to this region by linkage analysis:
CADASIL (cerebral autosomal dominant arteriopathy with
subcortical infarcts and leukoencephalopathy) between the
DNA markers D19S221 and D19S222, and FHM (familial
hemiplegic migraine) between D19S215 and D19S216. These
two diseases may be allelic disorders in which the same
gene is involved (Tournier-Lasserve, E., et al., Nature
Genet., 3, 256-259 (1993); Joutel, A., et al., Nature
Genet., 5, 40-45 (1993)].
Although no evidence is available to support
CKAP2 as a candidate gene for FHM or CADASIL, it is
. _,.., - . ..: r . . :...:.... .. . - - ...
. - . - . .--:.=,.. -. -:. . -:. . . ._:.-: . -._._:r_ ; =..---:_.-.--.- ----
.. . :. .. - _ _ -. .
CA 02458872 2004-03-23
-35-
conceivable that its mutation might lead to some or other
neurological disease.
By using the novel human CKAP2 gene of the
present invention as obtained in this example, it is
possible to detect the expression of said gene in various
tissues or produce the human CKAP2 gene in the manner of
genetic engineering. Through these, it becomes possible
to analyze the functions of the human CKAP2 system or
human CKAP2, which is involved in diverse activities
essential to cells, as mentioned above, to diagnose
various neurological diseases in which said system or
gene is involved, for example familial migraine, and to
screen out and evaluate a therapeutic or prophylactic
drug therefor.
Example 3
OTK27 gene
(1) OTK27 gene cloning and DNA sequencing
As a result of sequence analysis of cDNA clones
arbitraily selected from a human fetal brain cDNA library
in the same manner as in Example 1 (1) and database
searching, a cDNA clone, GEN-025F07, coding for a protein
highly homologous to NHP2, a yeast nucleoprotein
[Saccharomyces cerevisiae; Kolodrubetz, D. and Burgum,
A., YEAST, 7, 79-90 (1991)], was found and named OTK27.
Nucleoproteins are fundamental cellular consti-
õ;_ ._..
: .. . . . , :.. CA 02458872 2004-03-23
~.
-36-
tuents of chromosomes, ribosomes and so forth and are
thought to play an essential role in cell multiplication
and viability. The yeast nucleoprotein NHP2, a high-
mobility group (HMG)-like protein, like HMG, has
reportedly a function essential for cell viability
[Kolodrubetz, D. and Burgum, A., YEAST, 7, 79-90 (1991)].
The novel human gene, OTK27 gene, of the
present invention, which is highly homologous to the
above-mentioned yeast NHP2 gene, is supposed to be
similar in function.
The nucleotide sequence of said GEN-025F07
clone was found to comprise 1493 nucleotides, as shown
under SEQ ID NO:9, and contain an open reading frame
comprising 384 nucleotides, as shown under SEQ ID NO:8,
coding for an amino acid sequence comprising 128 amino
acid residues, as shown under SEQ ID NO:7. The
initiation codon was located at nucleotides 95-97 of the
sequence shown under SEQ ID N0:9, and the termination
codon at nucleotides 479-481.
At the amino acid level, the OTK27 protein was
highly homologous (38%) to NHP2. It was 83% identical
with the protein deduced from the cDNA from Arabidonsis
thaliana; Newman, T., unpublished; GENEMBL Accession No.
T14197).
(2) Northern blot analysis
_._..<_. : ~ .
CA 02458872 2004-03-23
-37-
For examining the expression of human OTK27
mRNA in normal human tissues, the insert in the OTK27
cDNA was amplified by PCR, the PCR product was purified
and labeled with [32P]-dCTP (random-primed DNA labeling
kit, Boehringer Mannheim), and Northern blotting was
performed using the labeled product as a probe in the
same manner as in Example 1 (2).
As a result of the Northern blot analysis, two
bands corresponding to possible transcripts from this
gene were detected at approximately 1.6 kb and 0.7 kb.
Both sizes of transcript were expressed in all normal
adult tissues examined. However, the expression of the
0.7 kb transcript was significantly reduced in brain and
was of higher levels in heart, skeletal muscle and
testicle than in other tissues examined.
For further examination of these two
transcripts, eleven cDNA clones were isolated from a
testis cDNA library and their DNA sequences were
determined in the same manner as in Example 1 (1).
As a result, in six clones, the sequences were
found to be in agreement with that of the 0.7 kb
transcript, with a poly(A) sequence starting at around
the 600th nucleotide, namely at the 598th nucleotide in
two of the six clones, at the 606th nucleotide in three
clones, and at the 613th nucleotide in one clone.
. -. .... ,..... .
-----------------
CA 02458872 2004-03-23
~.
-38-
In these six clones, the "TATAAA" sequence was
recognized at nucleotides 583-588 as a probable poly(A)
signal. The upstream poly(A) signal "TATAAA" of this
gene was recognized as little influencing in brain and
more effective in the three tissues mentioned above than
in other tissues. The possibility was considered that
the stability of each transcript vary from tissue to
tissue.
Results of zoo blot analysis indicated that
this gene is well conserved also in other vertebrates.
Since this gene is expressed ubiquitously in normal adult
tissues and conserved among a wide range of species, the
gene product is likely to play an important physiological
role. The evidence that yeasts lacking in NHP2 are
nonviable suggests that the human homolog may also be
essential to cell viability.
(3) Chromosomal localization of OTK27 by direct R-
banding FISH
One cosmid clone corresponding to the cDNA
OTK27 was isolated from a total human genomic cosmid
library (5-genome equivalent) using the OTK27 cDNA insert
as a probe and subjected to FISH in the same manner as in
Example 1 (3) for chromosomal localization of OTK27.
As a result, two distinct spots were observed
on the chromosome band 12q24.3.
CA 02458872 2004-03-23
~
-39-
The OTK27 gene of the present invention can be
used in causing expression thereof and detecting the
OTK27 protein, a human nucleoprotein, and thus can be
utilized in the diagnosis and pathologic studies of
various diseases in which said protein is involved and,
because of its involvement in cell proliferation and
differentiation, in screening out and evaluating
therapeutic and preventive drugs for cancer.
Example 4
OTK18 gene
(1) OTK18 gene cloning and DNA sequencing
Zinc finger proteins are defined as constituing
a large family of transcription-regulating proteins in
eukaryotes and carry evolutionally conserved structural
motifs [Kadonaga, J. T., et al., Cell, 51, 1079-1090
(1987); Klung, A. and Rhodes, D., Trends Biol. Sci., 12,
464-469 (1987); Evans, R. M. and Hollenberg, S. M., Cell,
52, 1-3 (1988)].
The zinc finger, a loop-like motif formed by
the interaction between the zinc ion and two residues,
cysteine and histidine residues, is involved in the
sequence-specific binding of a protein to RNA or DNA.
The zinc finger motif was first identified within the
amino acid sequence of the Xenopus transcription factor
IIIA [Miller, J., et al., EMBO J., 4, 1609-1614 (1986)].
CA 02458872 2004-03-23
-40-
The C2H2 finger motif is in general tandemly
repeated and contains an evolutionally conserved inter-
vening sequence of 7 or 8 amino acids. This intervening
stretch was first identified in the Kruppel segmentation
gene of Drosophila [Rosenberg, U. B., et al., Nature,
319, 336-339 (1986)]. Since then, hundreds of C2H2 zinc
finger protein-encoding genes have been found in
vertebrate genomes.
As a result of sequence analysis of cDNA clones
arbitrarily selected from a human fetal brain cDNA
library in the same manner as in Example 1 (1). and
database searching, several zinc finger structure-
containing clones were identified and, further, a clone
having a zinc finger structure of the Kruppel type was
f ound .
Since this clone lacked the 5' portion of the
transcript, plaque hybridization was performed with a
fetal brain cDNA library using, as a probe, an appro-
ximately 1.8 kb insert in the cDNA clone, whereby three
clones were isolated. The nucleotide sequences of these
were determined in the same manner as in Example 1 (1).
Among the three clones, the one having the
largest insert spans 3,754 nucleotides including an open
reading frame of 2,133 nucleotides coding for 711 amino
acids. It was found that said clone contains a novel
. . .- ......- -- a ..... . _ .: _.. . _.
= -. ~.- _ =....._ ' ..-..:1'_. -. ' . - - - . . . - _--_- -- .. . .. . . .. .
. .. . . ._- _.. . _. .. . . ...._. .... . .- -. . =. -' ~
CA 02458872 2004-03-23
~
-41-
human gene coding for a peptide highly homologous in the
zinc finger domain to those encoded by human ZNF41 and
the Drosophila Kruppel gene. This gene was named OTK18
gene (derived from the clone GEN-076C09).
The nucleotide sequence of the cDNA clone of
the OTK18 gene is shown under SEQ ID N0:12, the coding
region-containing nucleotide sequence under SEQ ID NO:11,
and the predicted amino acid sequence encoded by said
OTK18 gene under SEQ ID NO:10.
It was found that the amino acid sequence of
OTK18 as deduced from SEQ ID N0:12 contains 13 finger
motifs on its carboxy side.
(2) Comparison with other zinc finger motif-containing
genes
Comparison among OTK18, human ZNF41 and the
Drosophila Kruppel gene revealed that each finger motif
is for the most part conserved in the consensus sequence
CXECGKAFXQKSXLX2HQRXH.
Comparison of the consensus sequence of the
zinc finger motifs of OTK18 with those of human ZNF41 and
the Drosophila Kruppel gene revealed that the Kruppel
type motif is well conserved in the OTK18-encoded
protein. However, the sequence similarities were limited
to zinc finger domains and no significant homologies were
found with regard to other regions.
_ . - ..Y... .._ _. , __ . ..... ...t... . . . -; : - . . _. _,....... . _ ...-
,. .
:-:.... .-r __......_"_ __'= ___...-- .- . _..._ ___ __ __ _ _ _ _ _...___
r.:_>-.. :. . .. . ...............w.,...._....,....._..L..rs._.._. ...-_
.. ._. _... _:__:_'____ ~= -
CA 02458872 2004-03-23
-42-
The zinc finger domain interacts specifically
with the target DNA, recognizing an about 5 bp sequence
to thereby bind to the DNA helix [Rhodes, D. and Kiug,
A., Cell, 46, 123-132 (1986)].
Based on the idea that, in view of the above,
the multiple module (tandem repetitions of zinc finger)
can interact with long stretches of DNA, it is presumable
that the target DNA of this gene product containing 13
repeated zinc finger units would be a DNA fragment with a
length of approximately 65 bp.
(3) Northern blot analysis
Northern blot analysis was performed as
described in Example 1 (2) for checking normal human
tissues for expression of the human OTK18 mRNA therein by
amplifying the insert of the OTK18 cDNA by PCR, purifying
the PCR product, labeling the same with [32P]-dCTP
(random-primed DNA labeling kit, Boehringer Mannheim) and
using an MTN blot with the labeled product as a probe.
The results of Northern blot analysis revealed
that the transcript of OTK18 is approximately 4.3 kb long
and is expressed ubiquitously in various normal adult
tissues. However, the expression level in the liver and
in peripheral blood lymphocytes seemed to be lower than
in other organs tested. '
(4) Cosmid cloning and chromosomal localization by
.., . ,...,........ ,,.. r. .. . . .-. ._.. _, ~.-:. -. . .::-:. .. --._. ., .
, . . .
CA 02458872 2004-03-23
-43-
direct R-banding FISH
Chromosomal localization of OTK18 was carried
out as described in Example 1 (3).
As a result, complete twin spots were
identified with 8 samples while 23 samples showed an
incomplete signal or twin spots on either or both
homologs. All signals appeared at the q13.4 band of
chromosome 19. No twin spots were observed on any other
chromosomes.
The results of FISH thus revealed that this
gene is localized on chromosomal band 19q13.4. This
region is known to contain many DNA segments that
hybridize with oligonucleotides corresponding to zinc
finger domains [Hoovers, J. M. N., et al., Genomics, 12,
254-263 (1992)]. In addition, at least one other gene
coding for a zinc finger domain has been identified in
this region [Marine, J.-C., et al., Genomics, 21, 285-286
(1994)].
Hence, the chromosome 19q13 is presumably a
site of grouping of multiple genes coding for
transcription-regulating proteins.
When the novel human OTK18 gene provided by
this example is used, it becomes possible to detect
expression of said gene in various tissues and produce
the human OTK18 protein in the manner of genetic
CA 02458872 2004-03-23
-44-
engineering. Through these, it is possible to analyze
the functions of the human transcription regulating
protein gene system or human transcription regulating
proteins, which are deeply involved in diverse activities
fundamental to cells, as mentioned above, to diagnose
various diseases with which said gene is associated, for
example malformation or cancer resulting from a
developmental or differentiation anomaly, and mental or
nervous disorder resulting from a developmental anomaly
in the nervous system, and further to screen out and
evaluate therapeutic or prophylactic drugs for these
diseases.
ExamDle 5
Genes encoding human 26S proteasome constituent P42
protein and P27 protein
(1) Cloning and DNA sequencing of genes respectively
encoding human 26S proteasome constituent P42
protein and P27 protein
Proteasome, which is a multifunctional
protease, is an enzyme occurring widely in eukaryotes
from yeasts to humans and decomposing ubiquitin-binding
proteins in cells in an energy-dependent manner.
=Structurally, said proteasome is constituted of 20S
proteasome composed of various constituents with a
molecular weight of 21 to 31 kilodaltons and a group of
. .............. ...,.
_- -- ~ .., --.. _ - -- -- ------
CA 02458872 2004-03-23
-45-
PA700 regulatory proteins composed of various
constituents with a molecular weight of 30 to 112
kilodaltons and showing a sedimentation coefficient of
22S and, as a whole, occurs as a macromolecule with a
molecular weight of about 2 million daltons and a
sedimentation coefficient of 26S [Rechsteiner, M., et
al., J. Biol. Chem., 268, 6065-6068 (1993); Yoshimura,
T., et al., J. Struct. Biol., 111, 200-211 (1993);
Tanaka, K., et al., New Biologist, 4, 173-187 (1992)].
Despite structural and mechanical analyses
thereof, the whole picture of proteasome is not yet fully
clear. However, according to studies using yeasts and
mice in the main, it reportedly has the functions
mentioned below and its functions are becoming more and
more elucidated.
The mechanism of energy-dependent proteolysis
in cells starts with selection of proteins by ubiquitin
binding. It is not 20S proteasome but 26S proteasome
that has ubiquitin-conjugated protein decomposing
activity which is ATP-dependent [Chu-Ping et al., J.
Biol. Chem., 269, 3539-3547 (1994)]. Hence, human 26S
proteasome is considered to be useful in elucidating the
mechanism of energy-dependent proteolysis.
Factors involved in the cell cycle regulation
are generally short in half-life and in many cases they
CA 02458872 2004-03-23
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are subject to strict quantitative control. In fact, it
has been made clear that the oncogene products Mos, Myc,
Fos and so forth can be decomposed by 26S proteasome in
an energy- and ubiquitin-dependent manner [Ishida, N., et
al., FEBS.Lett., 324, 345-348 (1993); Hershko, A. and
Ciechanover, A., Annu. Rev. Biochem., 61, 761-807
(1992)] and the importance of proteasone in cell cycle
control is being recognized.
Its importance in the immune system has also
been pointed out. It is suggested that proteasome is
positively involved in class I major histocompatible
complex antigen presentation [Michalek, M. T., et al.,
Nature, 363, 552-554 (1993)] and it is further suggested
that proteasome may be involved in Alzheimer disease,
since the phenomena of abnormal accumulation of
ubiquitin-conjugated proteins in the brain of patients
with Alzheimer disease [Kitaguchi, N., et al., Nature,
361, 530-532 (1988)]. Because of its diverse functions
such as those mentioned above, proteasome attracts
attention from the viewpoint of its utility in the
diagnosis and treatment of various diseases.
A main function of 26S proteasome is ubiquitin-
conjugated protein decomposing activity. In particular,
it is known that cell cycle-related gene products such as
oncogene products and cyclins, typically c-Myc, are
CA 02458872 2004-03-23
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degraded via ubiquitin-dependent pathways. It has also
been observed that the proteasome gene is expressed
abnormally in liver cancer cells, renal cancer cells,
leukemia cells and the like as compared with normal cells
[Kanayama, H., et al., Cancer Res., 51, 6677-6685 (1991)]
and that proteasome is abnormally accumulated in tumor
cell nuclei. Hence, constituents of proteasome are
expected to be useful in studying the mechanism of such
canceration and in the diagnosis or treatment of cancer.
Also, it is known that the expression of
proteasome is induced by interferon y and so on and is
deeply involved in antigen presentation in cells [Aki,
M., et al., J. Biochem., 115, 257-269 (1994)]. Hence,
constituents of human proteasome are expected to be
useful in studying the mechanism of antigen presentation
in the immune system and in developing immunoregulating
drugs.
Furthermore, proteasome is considered to be
deeply associated with ubiquitin abnormally accumulated
in the brain of patients with Alzheimer disease. Hence,
it is suggested that constituents of human proteasome
should be useful in studying the cause of Alzheimer
disease and in the treatment of said disease.
In addition to the utilization of expectedly
multifunctional proteasome as such in the above manner,
- - ......w......a....i..i-tY ..+rn+M1/~..-_...Jw.~ rO::i[Y.tl...'li-
:J"'Yr'..n..._ w.-ww..w+='ru'.s.a _:::J s..'f._ lJY :ff1=f.1M/f=~r.r.r.J..-._v-
.rw...._ ... .. .. ... .... ..: ... .. !Ti...-ir,.v....
CA 02458872 2004-03-23
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it is probably possible to produce antibodies using
constituents of proteasome as antigens and use such
antibodies in diagnosing various diseases by immunoassay.
Its utility in this field of diagnosis is thus also a
focus of interest.
Meanwhile, a protein having the characteristics
of human 26S proteasome is disclosed, for example in
Japanese Unexamined Patent Publication No. 292964/1993
and rat proteasome constituents are disclosed in Japanese
Unexamined Patent Publication Nos. 268957/1993 and
317059/1993. However, no human 26S proteasome
constituents are known. Therefore, the present inventors
made_a further search for human 26S proteasome
constituents and successfully obtained two novel human
26S proteasome constituents, namely human 26S proteasome
constituent P42 protein and human S26 proteasome
constituent P27 protein, and performed cloning and DNA
sequencing of the corresponding genes in the following
manner.
(1) Purification of human 26S proteasome constituents
P42 protein and P27 protein
Human proteasome was purified using about 100 g
of fresh human kidney and following the method of purify-
ing human proteasome as described in Japanese Unexamined
Patent Publication No. 292964/1993, namely by column
CA 02458872 2006-09-22
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TM
chromatography using BioGel A-1.5 m (5 x 90 cm, Bio-Rad),
hydroxyapatite (1.5 x 15 cm, Bio-Rad) and Q-Sepharose
(1.5 x 15 cm, Pharmacia) and glycerol density gradient
centrifugation.
The thus-obtained human proteasome was
subjected to reversed phase high performance liquid
chromatography (HPLC) using a Hitachi model'L6200 HPLC
TM
system. A Shodex RS Pak D4-613 (0.6 x 15 cm, Showa
Denko) was used and gradient elution was performed with
the following two solutions:
First solution: 0.06% trifluoroacetic acid;
Second solution: 0.05$ trifluoroacetic acid, 70t
acetonitrile.
An aliquot of each eluate fraction was
subjected to 8.5$ SDS-polyacrylamide electrophoresis
under conditions of reduction with dithiothreitol. The
P42 protein and P27 protein thus detected were isolated
and purified.
The purified P42 and P27 proteins were respec-
tively digested with 1 pg of trypsin in 0.1 M Tris buffer
(pH 7.8) containing 2 M urea at 37 C for 8 hours and the
partial peptide fragments obtained were separated by
reversed phase HPLC and their sequences were determined
by Edman degradation. The results obtained are as shown
below in Table 2.
....----- _ _ _ _ _ ....._v.._- ._. ....... .. ... ... . . _ _ --- - ._.-
_._._._._... _ ____---------._,
CA 02458872 2004-03-23
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Table 2
Partial protein Amino acid sequence
P42 (1) VLNISLW
(2) TLMELLNQMDGFDTLHR
(3) AVSDFVVSEYXMXA
(4) EVDPLVYNX
(5) HGEIDYEAIVK
(6) LSXGFNGADLRNVXTEAGMFAIXAD
(7) MIMATNRPDTLDPALLRPGXL
(8) IHIDLPNEQARLDILK
(9) ATNGPRYVVVG
(10) EIDGRLK
(11) ALQSVGQIVGEVLK
(12) ILAGPITK
(13) XXVIELPLTNPELFQG
(14) VVSSSLVDK
(15) ALQDYRK
(16) EHREQLK
(17) KLESKLDYKPVR
P27 (1) LVPTR
(2) AKEEEIEAQIK
(3) ANYEVLESQK
(4) VEDALHQLHAR
(5) DVDLYQVR
(6) QSQGLSPAQAFAK
(7) AGSQSGGSPEASGVTVSDVQE
(8) GLLGXNIIPLQR
Y- r ^<n inn+ av.- : . --.: -.:"= ="='-= :-' ' -. ==r _...- CA 02458872 2004-
03-23 -...._ .. _.. ... ... _.- _. . .. 'rr ' w ~.:.. .
-51-
(2) cDNA library screening, clone isolation and cDNA
nucleotide sequence determination
As mentioned in Example 1 (1), the present
inventors have a database comprising about 30,000 cDNA
data as constructed based on large-scale DNA sequencing
using human fetal brain, arterial blood vessel and
placenta cDNA libraries.
Based on the amino acid sequences obtained as
mentioned above in (1), computer searching was performed
with the FASTA program (search for homology between said
amino acid sequences and the amino acid sequences
estimated from the database). As regards P42, a clone
(GEN-331G07) showing identity with regard to two amino
acid sequences [(2) and (7) shown in table 2] was
screened out and, as regards P27, a clone (GEN-163D09)
showing identity with regard to two amino acid sequences
[(1) and (8) shown in Table 2] was found.
For each of these clones, the 5' side sequence
was determined by 5' RACE and the whole sequence was
determined, in the same manner as in Example 2 (2).
As a result, it was revealed that the above-
mentioned P42 clone GEN-331G07 comprises a 1,566-
nucleotide sequence as shown under SEQ ID N0:15,
inclusive of a 1,167-nucleotide open reading frame as
shown under SEQ ID N0:14, and that the amino acid
CA 02458872 2004-03-23
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sequence encoded thereby is the one shown under SEQ ID
N0:13 and comprises 389 amino acid residues.
The results of computer homology search
revealed that the P42 protein is significantly homologous
to the AAA (ATPase associated with a variety of cellular
activities) protein family (e.g. P45, TBP1, TBP7, S4,
MSS1, etc.). It was thus suggested that it is a new
member of the AAA protein family.
As for the P27 clone GEN-163D09, it was
revealed that it comprises a 1,128-nucleotide sequence as
shown under SEQ ID NO:18, including a 669-nucleotide open
reading frame as shown under SEQ ID NO:17 and that the
amino acid sequence encoded thereby is the one shown
under SEQ ID N0:16 and comprises 223 amino acid residues.
As regards the P27 protein, homology search
using a computer failed to reveal any homologous gene
among public databases. Thus, the gene in question is
presumably a novel gene having an unknown function.
Originally, the above-mentioned P42 and P27
gene products were both purified as regulatory subunit
components of proteasome complex. Therefore, these are
expected to play an important role in various biological
functions through proteolysis, for example a role in
energy supply through decomposition of ATP and, hence,
they are presumably useful not only in studying the
------------------------------ -------------.
CA 02458872 2004-03-23
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function of human 26S proteasome but also in the
diagnosis and treatment of various diseases caused by
lowering of said biological functions, among others.
Example 6
BNAP gene
(1) BNAP gene cloning and DNA sequencing
The nucleosome composed of DNA and histone is a
fundamental structure constituting chromosomes in
eukaryotic cells and is well conserved over borders among
species. This structure is closely associated with the
processes of replication and transcription of DNA.
However, the nucleosome formation is not fully understood
as yet. Only certain specific factors involved in
nucleosome assembly (NAPs) have been identified. Thus,
two acidic proteins, nucleoplasmin and N1, are already
known to facilitate nucleosome construction
[Kleinschmidt, J. A., et al., J. Biol. Chem., 260, 1166-
1176 (1985); Dilworth, S. M., et al., Cell, 51, 1009-1018
(1987)].
A yeast gene, NAP-I, was isolated using a mono-
clonal antibody and recombinant proteins derived
therefrom were tested as to whether they have nucleosome
assembling activity in vivo.
More recently, a mouse NAP-I gene, which is a
mammalian homolog of the yeast NAP-I gene was cloned
._._
------------.. ------ _-_---.-------- ---- ...... ........
CA 02458872 2004-03-23
-54-
(Okuda, A.; registered in database under the accession
number D12618). Also cloned were a mouse gene, DN38
[Kato, K., Eur. J. Neurosci., 2, 704-711 (1990)] and a
human nucleosome assembly protein (hNRP) [Simon, H. U.,
et al., Biochem. J., 297, 389-397 (1994)]. It was shown
that the hNRP gene is expressed in many tissues and is
associated with T lymphocyte proliferation.
The present inventors performed sequence
analysis of cDNA clones arbitrarily chosen from a human
fetal brain cDNA library in the same manner as in Example
1 (1), followed by searches among databases and, as a
result, made it clear that a 1,125-nucleotide cDNA clone
(free of poly(A)), GEN-078D05, is significantly
homologous to the mouse NAP-I gene, which is a gene for a
nucleosome assembly protein (NAP) involved in nucleosome
construction, a mouse partial cDNA clone, DN38, and hNRP.
Since said clone GEN-078D05 was lacking in the
5' region, 5' RACE was performed in the same manner as in
Example 2 (2) to obtain the whole coding region. For
this 5' RACE, primers P1 and P2 respectively having the
nucleotide sequences shown below in Table 3.
Table 3
Primer Nucleotide sequence
Primer P1 5'-TTGAAGAATGATGCATTAGGAACCAC-3'
Primer P2 5'-CACTCGAGTGGCTGGATTTCAATTTCTCCAGTAG-3'
- . ..__ ....._...,..-._ .:. ..
------....-..... - ........ .. . .. . ..., : :: _-r,. . ,.~. ~~, ,~. ~~ _._.
.... ...
CA 02458872 2004-03-23
-55-
After the first 5' RACE, a single band
corresponding to a sequence length of 1,300 nucleotides
was obtained. This product was inserted into pT7Blue(R)
T-Vector and several clones appropriate in insert size
were selected.
Ten 5' RACE clones obtained from two
independent PCR reactions were sequenced and the longest
clone GEN-078D05TA13 (about 1,300 nucleotides long) was
further analyzed.
Both strands of the two overlapping cDNA clones
GEN-078D05 and GEN-078D05TA13 were sequenced, whereby it
was confirmed that the two clones did not yet cover the
whole coding region. Therefore, a further second 5' RACE
was carried out. For the second 5' RACE, two primers, P3
and P4, respectively having the sequences shown below in
Table 4 were used.
Table 4
Primer Nucleotide sequence
Primer P3 5'-GTCGAGCTAGCCATCTCCTCTTCG-3'
Primer P4 5'-CATGGGCGACAGGTTCCGAGACC-3'
A clone, GEN-078D0508, obtained by the second
5' RACE was 300 nucleotides long. This clone contained
an estimable initiation codon and three preceding in-
frame termination codons. From these three overlapping
clones, it became clear that the whole coding region
CA 02458872 2004-03-23
-56-
comprises 2,636 nucleotides. This gene was named brain-
specific nucleosome assembly protein (BNAP) gene.
The BNAP gene contains a 1,518-nucleotide open
reading frame shown under SEQ ID N0:20. The amino acid
encoded thereby comprises 506 amino acid residues; as
shown under SEQ ID N0:19, and the nucleotide sequence of
the whole cDNA clone of BNAP is as shown under SEQ ID
NO:21.
As shown under SEQ ID NO:21, the 5' noncoding
region of said gene was found to be generally rich in GC.
Candidate initiation codon sequences were found at
nucleotides Nos. 266-268, 287-289 and 329-331. These
three sequences all had well conserved sequences in the
vicinity of the initiation codons [Kozak, M., J. Biol.
Chem., 266, 19867-19870 (1991)].
According to the scanning model, the first ATG
(nucleotides Nos. 266-268) of the cDNA clone may be the
initiation codon. The termination codon was located at
nucleotides Nos. 1784-1786.
The 3' noncoding redion was generally rich in
AT and two polyadenylation signals (AATAAA) were located
at nucleotides Nos. 2606-2611 and 2610-2615,
respectively.
The longest open reading frame comprised 1,518
nucleotides coding for 506 amino acid residues and the
CA 02458872 2004-03-23
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calculated molecular weight of the BNAP gene product was
57,600 daltons.
Hydrophilic plots indicated that BNAP is very
hydrophilic, like other NAPs.
For recombinant BNAP expression and
purification and for eliminating the possibility that the
BNAP gene sequence might give three chimera clones in the
step of 5' RACE, RT-PCR was performed using a sequence
comprising nucleotides Nos. 326-356 as a sense primer and
a sequence comprising nucleotides Nos. 1758-1786 as an
antisenses primer.
As a result, a single product of about 1,500 bp
was obtained and it was thus confirmed that said sequence
is not a chimera but a single transcript.
(2) Comparison between BNAP and NAPs
The amino acid sequence deduced from BNAP
showed 46$ identity and 65% similarity to hNRP.
The deduced BNAP gene product had motifs
characteristic of the NAPs already reported and of BNAP.
In general, half of the C terminus was well conserved in
humans and yeasts.
The first motif (domain I) is KGIPDYWLI (corres
ponding to amino acid residues Nos. 309-317). This was
observed also in hNRP (KGIPSFWLT) and in yeast NAP-I
(KGIPEFWLT).
----=- - - --=--._~ .............. _,... > ...__.._...__..._~s__..=- CA
02458872 2004~ 03=23, .,........,.,-.Y-~---
...M........,._,...w...,..<..,..Y..,....,.x..r.,~,....Rr
-58-
The second motif (domain II) is ASFFNFFSPP
(corresponding to amino acid residues Nos. 437-446) and
this was expressed as DSFFNFFAPP in hNRP and as ESFFNFFSP
in yeast NAP-I.
These two motifs were also conserved in the
deduced mouse NAP-I and DN38 peptides. Both conserved
motifs were each a hydrophilic cluster, and the Cys in
position 402 was also found conserved.
Half of the N terminus had no motifs strictly
conserved from yeasts to mammalian species, while motifs
conserved among mammalian species were found.
For instance, HDLERKYA (corresponding to amino
acid residues Nos. 130 to 137) and IINAEYEPTEEECEW
(corresponding to amino acid residues Nos. 150-164),
which may be associated with mammal-specific functions,
were found strictly conserved.
NAPs had acidic stretches, which are believed
to be readily capable of binding to histone or other
basic proteins. All NAPs had three acidic stretches but
the locations thereof were not conserved.
BNAP has no such three acidic stretches but,
instead, three repeated sequences (corresponding to amino
acid residues Nos. 194-207, 208-221 and 222-235) with a
long acidic cluster, inclusive of 41 amino acid residues
out of 98 amino acid residues, the consensus sequence
CA 02458872 2004-03-23
-59-
being ExxKExPEVKxEEK (each x being a nonconserved, mostly
hydrophobic, residue).
Furthermore, it was revealed that the BNAP
sequence had several BNAP-specific motifs. Thus, an
extremely serine-rich doamin (corresponding to amino acid
residues Nos. 24-72) with 33 (67%) of 49 amino acid
residues being serine residues was found in the N-
terminus portion. On the nucleic acid level, they were
reflected as incomplete repetitions of AGC.
Following this serine-rich region, there
appeared a basic domain (corresponding to amino acid
residues Nos. 71-89) comprising 10 basic amino acid
residues among 19 residues.
BNAP is supposed to be localized in the
nucleus. Two possible signals localized in the nucleus
were observed (NLSs). The first signal was found in the
basic domain of BNAP and its sequence YRKKR (corres-
ponding to amino acid residues Nos. 75-79) was similar to
NLS (GRKKR) of Tat of HIV-1. The second signal was
located in the C terminus and its sequence KKYRK
(corresponding to amino acid residues Nos. 502-506) was
similar to NLS (KKKRK) of,the large T antigen of SV40.
The presence of these two presumable NLSs suggested the
localization of BNAP in the nucleus. However the
possibility that other basic clusters might act as NLSs
CA 02458872 2004-03-23
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could not be excluded.
BNAP has several phosphorylation sites and the
activity of BNAP may be controlled through phosphoryla-
tion thereof.
(3) Northern blot analysis
Northern blot analysis was performed as
described in Example 1 (2). Thus, the clone GEN-
078D05TA13 (corresponding to nucleotides Nos. 323 to 1558
in the BNAP gene sequence) was amplified by PCR, the PCR
product was purified and labeled with [32P]-dCTP (random-
primed DNA labeling kit, Boehringer Mannheim), and the
expression of BNAP mRNA in normal human tissues was
examined using an MTN blot with the labeled product as a
probe.
As a result of Northern blot analysis, a 3.0 kb
transcript of BNAP was detected (8-hour exposure) in the
brain among eight human adult tissues tested, namely
heart, brain, placenta, lung, liver, skeletal muscle,
kidney and pancreas and, after longer exposure (24
hours), a dim band of the same size was detected in the
heart.
BNAP was found equally expressed in several
sites of brain tested whereas, in other tissues, no
signal was detected at all even after 72 hours of
exposure. hNRP mRNA was found expressed everywhere in
_.. _. _. . _,. . ,__ - . - .. .., __... _. _ . . ... ,..,. : ...,. .,. _:_:
:;:::-:.:.:, =. ...._ .... . - -- ._ .. ___._.. ... . ....__--_._. .........
_. . _ _ .
... . . .... . = ..-.. . _ -. :. : .-.:..- .... : . . .. .... .-. - - .... -..
..... ...r.,...,._._:r.._ _...=_. . . ...:..: --=---' -_
CA 02458872 2004-03-23
-61-
the human tissues tested whereas the expression of BNAP
mRNA was tissue-specific.
(4) Radiation hubrid mapping
Chromosomal mapping of the BNAP clone was
performed by means of radiation hibrid mapping [Cox, D.
R., et al., Science, 250, 245-250 (1990)].
Thus, a total human genome radiation hybrid
clone (G3RH) panel was purchased from Research Genetics,
Inc., AL, USA and PCR was carried out for chromosomal
mapping analysis according to the product manual using
two primers, Al and A2, respectively having the
nucleotide sequences shown in Table 5.
Table 5
Primer Nucleotide sequence
Al primer 5'-CCTAAAAAGTGTCTAAGTGCCAGTT-3'
A2 primer 5'-TCAGTGAAAGGGAAGGTAGAACAC-3'
The results obtained were analyzed utilizing
softwares usable on the Internet [Boehnke, M., et al.,
Am. J. Hum. Genet., 46, 581-586 (1991)].
As a result, the BNAP gene was found strongly
linked to the marker DXS990 (LOD = 1000, cR8000 = -0.00).
Since DXS990 is a marker localized on the chromosome
Xq21.3-q22, it was established that BNAP is localized to
the chromosomal locus Xq21.3-q22 where genes involved in
several signs or symptoms of X-chromosome-associated
CA 02458872 2004-03-23
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mental retardation are localized.
The nucleosome is not only a fundamental
chromosomal structural unit characteristic of eukaryotes
but also a gene expression regulating unit. Several
results indicate that genes with high transcription
activity are sensitive to nuclease treatment, suggesting
that the chromosome structure changes with the
transcription activity [Elgin, S. C. R., J. Biol. Chem.,
263, 19259-19262 (1988)].
NAP-I has been cloned in yeast, mouse and human
and is one of the factors capable of promoting nucleosome
construction in vivo. In a study performed on their
sequences, NAPs containing the epitope of the specific
antibody 4A8 were detected in human, mouse, frog,
Drosophila and yeast (Saccharomyces cerevisiae) [Ishimi,
Y., et al., Eur. J. Biochem., 162, 19-24 (1987)].
In these experiments, NAPs, upon SDS-PAGE
analysis, electrophoretically migrated to positions
corresponding to a molecular weight between 50 and 60
kDa, whereas the recombinant BNAP slowly migrated to a
position of about 80 kDa. The epitope of 4A8 was shown
to be localized in the second, well-conserved,
hydrophobic motif. And, it was simultaneously shown that
the triplet FNF is important as a part of the epitope
(Fujii-Nakata, T., et al., J. Biol. Chem., 267, 20980-
CA 02458872 2004-03-23
-63-
20986 (1992)].
BNAP also contained this consensus motif in
domain II. The fact that domain II is markedly
hydrophobic and the fact that domain II can be recognized
by the immune system suggest that it is probably
presented on the BNAP surface and is possibly involved in
protein-protein interactions.
Domain I, too, may be involved in protein-
protein interactions. Considering that these are
conserved generally among NAPs, though to a relatively
low extent, it is conceivable that they must be essential
for nucleosome construction, although the functional
meaning of the conserved domains is still unknown.
The hNRP gene is expressed in thyroid gland,
stomach, kidney, intestine, leukemia, lung cancer,
mammary cancer and so on [Simon, H. U., et al., Biochem.
J., 297, 389-397 (1994)]. Like that, NAPs are expressed
everywhere and are thought to be playing an important
role in fundamental nucleosome formation.
BNAP may be involved in brain-specific
nucleosome formation and an insufficiency thereof may
cause neurological diseases or mental retardation as a
result of deviated functions of neurons.
BNAP was found strongly linked to a marker on
the X-chromosome q21.3-q22 where sequences involved in
------ . . ,.=:-.
CA 02458872 2004-03--23 ~
-64-
several symptoms of X-chromosome-associated mental
retardation are localized. This center-surrounding
region of X-chromosome was rich in genes responsible for
a-thalassemia, mental retardation (ATR-X) or some other
forms of mental retardation [Gibbons, R. J., et al.,
Cell, 80, 837-845 (1995)]. Like the analysis of the ATR-
X gene which seems to regulate the nucleosome structure,
the present inventors suppose that BNAP may be involved
in a certain type of X-chromosome-linked mental
retardation.
According to this example, the novel BNAP gene
is provided and, when said gene is used, it is possible
to detect the expression of said gene in various tissues
and to produce the BNAP protein by the technology of
genetic engineering. Through these, it is possible to
study the brain nucleosome formation deeply involved, as
mentioned above, in variegated activities essential to
cells as well as the functions of cranial nerve cells and
to diagnose various neurological diseases or mental
retardation in which these are involved and screen out
and evaluate drugs for the treatment or prevention of
such diseases.
Example 7
Human skeletal muscle-specific ubiquitin-conjugating
enzyme gene (UBE2G gene)
CA 02458872 2004-03-23
~
-65-
The ubiquitin system is a group of enzymes
essential for cellular processes and is conserved from
yeast to human. Said system is composed of ubiquitin-
activating enzymes (UBAs), ubiquitin-conjugating enzymes
(UBCs), ubiquitin protein ligases (UBRs) and 26S
proteasome particles.
Ubiquitin is transferred from the above-
mentioned UBAs to several UBCs, whereby it is activated.
UBCs transfer ubiquitins to target proteins with or
without the participation of UBRs. These ubiquitin-
conjugated target proteins are said to induce a number of
cellular responses, such as protein degradation, protein
modification, protein translocation, DNA repair, cell
cycle control, transcription control, stress responses,
etc. and immunological responses [Jentsch, S., et al.,
Biochim. Biophys. Acta, 1089, 127-139 (1991); Hershko, A.
and Ciechanover, A., Annu. Rev. Biochem., 61, 761-807
(1992); Jentsch, S., Annu. Rev. Genet., 26, 179-207
(1992); Ciechanover, A., Cell, 79, 13-21 (1994)].
UBCs are key components of this system and seem
to have distinct substrate specificities and modulate
different functions. For example, Saccharomyces
cerevisiae UBC7 is induced by cadmium and involved in
resistance to cadmium poisoning jJungmann, J., et al.,
Nature, 361, 369-371 (1993)]. Degradation of MAT-a2 is
CA 02458872 2004-03-23
~
-66-
also executed by UBC7 and UBC6 [Chen, P., et al., Cell,
74, 357-369 (1993)].
The novel gene obtained in this example is
UBC7-like gene strongly expressed in human skeletal
muscle. In the following, cloning and and DNA sequencing
thereof are described.
(1) Cloning and DNA sequencing of human skeletal muscle-
specific ubiquitin-conjugating enzyme gene (UBE2G
gene)
Following the same procedure as in Example 1
(1), cDNA clones were arbitrarily selected from a human
fetal brain cDNA library and subjected to sequence
analysis, and database searches were performed. As a
result, a cDNA clone, GEN-423A12, was found to have a
significantly high level of homology to the genes coding
for ubiquitin-conjugating enzymes (UBCs) in various
species.
Since said GEN-423A12 clone was lacking in the
5' side, 5' RACE was performed in the same manner as in
Example 2 (2) to obtain an entire coding region.
For said 5' RACE, two primers, P1 and P2,
respectively having the nucleotide sequences shown in
Table 6 were used.
..----_..~_:..__.. _-~,,::.-:.._, .. . .:..............:: ,,.,...,M:::-.r:.;:.
., ..._ _'.: --- ----._=._...__.._~:_. :.,._..._ .,. : . .
CA 02458872 2004-03-23
~
-67-
Table 6
Primer Nucleotide sequence
P1 primer 5'-TAATGAATTTCATTTTAGGAGGTCGG-3'
P2 primer 5'-ATCTTTTGGGAAAGTAAGATGAGCC-3'
The 5' RACE product was inserted into
pT7Blue(R) T-Vector and clones with an insert proper in
size were selected.
Four of the 5' RACE clones obtained from two
independent PCR reactions contained the same sequence but
were different in length.
By sequencing the above clones, the coding
sequence and adjacent 5'- and 3'-flanking sequences of
the novel gene were determined.
As a result, it was revealed that the novel
gene has a total length of 617 nucleotides. This gene
was named human skeletal muscle-specific ubiquitin-
conjugating enzyme gene (UBE2G gene).
To exclude the conceivable possibility that
this sequence was a chimera clone, RT-PCR was performed
in the same manner as in Example 6 (1) using the sense
primer to amplify said sequence from the human fetal
brain cDNA library. As a result, a single PCR product
was obtained, whereby it was confirmed that said sequence
is not a chimera one.
The UBE2G gene contains an open reading frame
CA 02458872 2004-03-23
-68-
of 510 nucleotides, which is shown under SEQ ID N0:23,
the amino acid sequence encoded thereby comprises 170
amino acid residues, as shown under SEQ ID N0:22, and the
nucleotide sequence of the entire UBE2G cDNA is as shown
under SEQ ID N0:24.
As shown under SEQ ID NO:24, the estimable
initiation codon was located at nucleotides Nos. 19-21,
corresponding to the first ATG triplet of the cDNA clone.
Since no preceding in-frame termination codon was found,
it was deduced that this clone contains the entire open
reading frame on the following grounds.
Thus, (a) the amino acid sequence is highly
homologous to S. cerevisiae UBC7 and said initiation
codon agrees with that of yeast-UBC7, supporting said ATG
as such. (b) The sequence AGGATGA is similar to the
consensus sequence (A/G)CCATGG around the initiation
codon [Kozak, M., J. Biol. Chem., 266, 19867-19870
(1991)].
(2) Comparison in amino acid sequence between UBE2G and
UBCs
Comparison in amino acid sequence between UBE2G
and UBCs suggested that the active site cystein capable
of binding to ubiquitin should be the 90th residue
cystein. The peptides encoded by these genes seem to
belong to the same family.
-_ _,.,_ .A=-.. .. .... ._. _...__.. -___-_._.__
CA 02458872 2004-03-23
~ r ,. =
-69-
(3) Northern blot analysis
Northern blot analysis was carried out as des-
cribed in Example 1 (2). Thus, the entire sequence of
UBE2G was amplified by PCR, the PCR product was purified
and labeled with [32P]-dCTP (random-primed DNA labeling
kit, Boehringer Mannheim) and the expression of UBE2G
mRNA in normal human tissues using the labeled product as
a probe. The membrane used was an MTN blot.
As a result of the Northern blot analysis, 4.4
kb, 2.4 kb and 1.6 kb transcripts could be detected in
all 16 human adult tissues, namely heart, brain,
placenta, lung, liver, skeletal muscle, kidney, pancreas,
spleen, thyroid gland, urinary bladder, testis, ovary,
small intestine, large intestine and peripheral blood
leukocye, after 18 hours of exposure. Strong expression
of these transcripts was observed in skeletal muscle.
(4) Radiation hybrid mapping
Chromosomal mapping of the UBE2G clone was per-
formed by radiation hybrid mapping in the same manner as
in Example 6 (4).
The primers Cl and C4 used in PCR for
chromosomal mapping analysis respectively correspond to
nucleotides Nos. 415-435 and nucleotides Nos. 509-528 in
the sequence shown under SEQ ID NO:24 and their
nucleotide sequences are as shown below in Table 7.
...._ -,..: ._- __.
.._. :.__- :- :.. ,:. .... - : : . . . . . ,_..__ _.~ - . _...------- ------- -
- ----------- -------------Y _,. .
------
CA 02458872 2004-03-23
.. .
-70-
Table 7
Primer Nucleotide sequence
Cl primer 5'-GGAGACTCACCTGCTAATGTT-3'
C4 primer 5'-CTCAAAAGCAGTCTCTTGGC-3'
As a result, the UBE2G gene was found linked to
the markers D1S446 (LOD = 12.52, cR8000 = 8.60) and
D1S235 (LOD = 9.14, cR8000 = 22.46). These markers are
localized to the chromosome bands 1q42.13-q42.3.
UBE2G was expressed strongly in skeletal muscle
and very weakly in all other tissues examined. All other
UBCs are involved in essential cellular functions, such
as cell cycle control, and those UBCs are expressed
ubiquitously. However, the expression pattern of UBE2G
might suggest a muscle-specific role thereof.
While the three transcripts differing in size
were detected, attempts failed to identify which
corresponds to the cDNA clone. The primary structure of
the UBE2G product showed an extreme homology to yeast
UBC7. On the other hand, nematode UBC7 showed strong
homology to yeast UBC7. It is involved in degradation of
.25 the repressor and further confers resistance to cadmium
in yeasts. The similarities among these proteins suggest
that they belong to the same family.
It is speculated that UBE2G is involved in
degradation of muscle-specific proteins and that a defect
_ _--------.___..~,-n.r.,.:,...........-._-_.._... _. ... _. _ .,.
.._,._...,.,......w___.._.._... _ , ._..- ... . .. . . .
CA 02458872 2004-03-23
~ .. r
-71-
in said gene could lead to such diseases as muscular
dystrophy. Recently, another proteolytic enzyme, calpain
3, was found to be responsible for limb-girdle muscular
dystrophy type 2A [Richard, I., et al., Cell, 81, 27-40
(1995)]. At the present, the chromosomal location of
UBE2G suggests no significant relationship with any
hereditary muscular disease but it is likely that a
relation to the gene will be unearthed by linkage
analysis in future.
In accordance with this example, the novel
UBE2G gene is provided and the use of said gene enables
detection of its expression in various tissues and
production of the UBE2G protein by the technology of
genetic engineering. Through these, it becomes possible
to study the degradation of muscle-specific proteins
deeply involved in basic activities variegated and
essential to cells, as mentioned above, and the functions
of skeletal muscle, to diagnose various muscular diseases
in which these are involved and further to screen out and
evaluate drugs for the treatment and prevention of such
diseases.
Example 8
TMP-2 gene
(1) TMP-2 gene cloning and DNA sequencing
Following the procedure of Example 1 (1), cDNA
...,..__. .... .., . ............... . ... . . . .... .....--=---- --_ _.. . .
. , . - . ... ... .
CA 02458872 2004-03-23
-72-
clones were arbitrarily selected from a human fetal brain
cDNA library and subjected to sequence analysis, and
database searches were performed. As a result, a clone
(GEN-092E10) having a cDNA sequence highly homologous to
a transmembrane protein gene (accession No.: U19878) was
found out.
Membrane protein genes have so far been cloned
in frog (Xenopus laevis) and human. These are considered
to be a gene for a transmembrane type protein having a
follistatin module and an epidermal growth factor (EGF)
domain (accession No.: U19878).
The sequence information of the above protein
gene indicated that the GEN-092E10 clone was lacking in
the 5' region, so that the a.gt10 cDNA library (human
fetal brain 5'-STRETCH PLUS cDNA; Clontech) was screened
using the GEN-092E10 clone as a probe, whereby a cDNA
clone containing a further 5' upstream region was
isolated.
Both strands of this cDNA clone were sequenced,
whereby the sequence covering the entire coding region
became clear. This gene was named TMP-2 gene.
The TMP-2 gene was found to contain an open
reading frame of 1,122 nucleotides, as shown under SEQ ID
N0:26, encoding an amino acid sequence of 374 residues,
as shown under SEQ ID N0:25. The nucleotide sequence of
,.._. .._ . ._ _,... _. ._ _
.......... ... .... .. . . _: _ _- _:,_w~x: - rr__ ;
~_._...._. ... _. -___ _.....<,.--... _ . - _~ : -
:_. .... __ . : .. _ >.:. -
CA 02458872 2004-03-23 ' _ _._....._ _
-73-
the entire TMP-2 cDNA clone comprises 1,721 nucleotides,
as shown under SEQ ID N0:27.
As shown under SEQ ID NO:27, the 5' noncoding
region was generally rich in GC. Several candidates for
the initiation codon were found but, according to the
scanning model, the 5th ATG of the cDNA clone (bases Nos.
368-370) was estimated as the initiation codon. The
termination codon was located at nucleotides Nos. 1490-
1492. The polyadenylation signal (AATAAA) was located at
nucleotides Nos. 1703-1708. The calculated molecular
weight of the TMP-2 gene product was 41,400 daltons.
As mentioned above, the transmembrane genes
have a follistatin module and an EGF domain. These
motifs were also found conserved in the novel human gene
of the present invention.
The TMP-2 gene of the present invention
presumably plays an important role in.cell proliferation
or intercellular communication, since, on the amino acid
level, said gene shows homology, across the EGF domain,
to TGF-a (transforming growth factor-a; Derynck, R., et
al., Cell, 38, 287-297 (1984)], beta-cellulin [Igarashi,
K. and Folkman, J., Science, 259, 1604-1607 (1993)],
heparin-binding EGF-like growth factor [Higashiyama, S.,
et al., Science, 251, 936-939 (1991)] and schwannoma-
derived growth factor (Kimura, H., et al., Nature, 348,
CA 02458872 2004-03-23
-74-
257-260 (1990)].
(2) Northern blot analysis
Northern blot analysis was carried out as des-
cribed in Example 1 (2). Thus, the clone GEN-092E10 was
amplified by PCR, the PCR product was purified and
labeled with [32P]-dCTP (random-primed DNA labeling kit,
Boehringer Mannheim), and the expression of TMP-2 mRNA in
normal human tissues was examined using an MTN blot with
the labeled product as a probe.
As a result, high levels of expression were
detected in brain and prostate gland. Said TMP-2 gene
mRNA was about 2 kb in size.
According to the present invention, the novel
human TMP-2 gene is provided and the use of said gene
makes it possible to detect the expression of said.gene
in various tissues or produce the human TMP-2 protein by
the technology of genetic engineering and, through these,
it becomes possible to study brain tumor and prostatic
cancer, which are closely associated with cell
proliferation or intercellular communication, as
mentioned above, to diagnose these diseases and to screen
out and evaluate drugs for the treatment and prevention
of such diseases.
Example 9
Human NPIK gene
CA 02458872 2004-03-23
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(1) Human NPIK gene cloning and DNA sequencing
' Following the procedures of Example 1 and
Example 2, cDNA clones were arbitrarily selected from=a
human fetal brain cDNA library and subjected to sequence
analysis, and database searches were performed. As a
result, two cDNA clones highly homologous to the gene
coding for an amino acid sequence conserved in
phosphatidylinositol 3 and 4 kinases [Kunz, J., et al.,
Cell, 73, 585-596 (1993)] were obtained. These were
named GEN-428B12c1 and GEN-428B12c2 and the entire
sequences of these were determined as in the foregoing
examples.
As a result, the GEN-428B12c1 cDNA clone and
the GEN-428B12c2 clone were found to have coding
sequences differing by 12 amino acid residues at the 5'
terminus, the GEN-428B12c1 cDNA clone being longer by 12
amino acid residues.
The GEN-428B12c1 cDNA sequence of the human
NPIK gene contained an open reading frame of 2,487
nucleotides, as shown under SEQ ID NO:32, encoding an
amino acid sequence comprising 829 amino acid residues,
as shown under SEQ ID NO:31. The nucleotide sequence of
the full-length cDNA clone comprised 3,324 nucleotides as
shown under SEQ ID NO:33.
The estimated initiation codon was located, as
...._ ... . .,,. . . . .. ...,. __ ...
CA 02458872 2004-03-23
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shown under SEQ ID NO:33, at nucleotides Nos. 115-117
corresponding to the second ATG triplet of the cDNA
clone. The termination codon was located at nucleotides
Nos. 2602-2604 and the polyadenylation signal (AATAAA) at
Nos. 3305-3310.
On the other hand, the GEN-428812c2 cDNA
sequence of the human NPIK gene contained an open reading
frame of 2,451 nucleotides, as shown under SEQ ID N0:29.
The amino acid sequence encoded thereby comprised 817
amino acid residues, as shown under SEQ ID N0:28. The
nucleotide sequence of the full-length cDNA clone
comprised 3,602 nucleotides, as shown under SEQ ID N0:30.
The estimated initiation codon was located, as
shown under SEQ ID N0:30, at nucleotides Nos. 429-431
corresponding to the 7th ATG triplet of the cDNA clone.
The termination codon was located at nucleotides Nos.
2880-2882 and the polyadenylation signal (AATAAA) at Nos.
3583-3588.
(2) Northern blot analysis
Northern blot analysis was carried out as des-
cribed in Example 1 (2). Thus, the entire sequence of
human NPIK was amplified by PCR, the PCR product was
purified and labeled with [32P]-dCTP (random-primed DNA
labeling kit, Boehringer Mannheim), and normal human
tissues were examined for expression of the human NPIK
CA 02458872 2006-09-22
-77-
= mRNA using the MTN blot membrane with the labeled product
as a probe. .
As a result, the expression of the human NPIK
gene was observed in 16 various human adult tissues
examined and an about 3.8 kb transcript and an about 5 kb
one could be detected.
Using primer A having the nucleotide sequence
shown below in Table 8 and containing the initiation
codon of the GEN-428B12c2 cDNA and primer B shown in
table 8 and containing the termination codon, PCR was
Tm
performed with Human Fetal Brain Marathon-Ready cDNA
(Clontech) as a template, and the nucleotide sequence of
the PCR product was determined.
Table 8
Primer Nucleotide sequence
Primer A 51-ATGGGAGATACAGTAGTGGAGC-3'
Primer B 5'-TCACATGATGCCGTTGGTGAG-3'
As a result, it was found that the human NPIK
mRNA expressed included one lacking in nucleotides Nos.
1060-1104 of the GEN-428B12c1 cDNA sequence (SEQ ID
N0:33) (amino acids Nos. 316-330 of the amino acid
sequence under SEQ ID N0:31) and one lacking in
nucleotides Nos. 1897-1911 of the GEN-428B12cl cDNA
sequence (SEQ ID N0:33) (amino acids Nos. 595-599 of the
amino acid sequence under SEQ ID N0:31).
_
.------------__
_._.,_:__ , ._... .. . . -.-:._.
CA 02458872 2004-03-23
1 M V
-78-
It was further revealed that polymorphism
existed in this gene (428B12cl.fasta), as shown below in
Table 9, in the region of bases Nos. 1941-1966 of the
GEN-428B12c1 cDNA sequence shown under SEQ ID NO:33,
whereby a mutant protein was encoded which resulted from
the mutation of IQDSCEITT (amino acid residues Nos. 610-
618 in the amino acid sequence (SEQ ID N0:31) encoded by
.GEN-428B12c1) into YKILVISA.
Table 9
1930 1940 1950 1959
TGGATCAAGCCAATACAAGATTCTTGTGAA
llillllllll IIIIlI1lllilllll
ICCATTTGGGAACAGGAGCGAGTGCCCCTTTGGATCAAGCC-ATACAAGATTCTTGTG--
1900 1910 1920 1930 1940 1950
1960 1970 1980
ATIACGACTGATAGTGGCATC
III II IIIIIIIIIIIIII
ATTTCGGCTGATAGTGGCATGATTGAACCAGTGGTCAATGCTGTGTCCATCCATCAGGTG
1960 1970 1980 1990 2000 2010
(3) Chromosomal mapping of human NPIK gene by FISH
Chromosomal mapping of the human NPIK gene was
carried out by FISH as described in Example 1 (3).
As a result, it was found that the locus of the
human NPIK gene is in the chromosomal position 1q21.1-
q21.3.
The human NPIK gene, a novel human gene, of the
present invention included two cDNAs differing in the 5'
region and capable of encoding 829 and 817 amino acid
_ ..._.,...:_,.:.. . ,__._.,........_
CA 02458872 2004-03-23
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residues, as mentioned above. In view of this and
further in view of the findings that the mRNA corres-
ponding to this gene includes two deletable sites and
there occurs polymorphism in a specific region corres-
ponding to amino acid residues Nos. 610-618 of the GEN-
428B12c1 amino acid sequence (SEQ ID N0:31), whereby a
mutant protein is encoded, it is conceivable that human
NPIK includes species resulting from a certain number of
combinations, namely human NPIK, deletion-containing
human NPIK, human NPIK mutant and/or deletion-containing
human NPIK mutant.
Recently, several proteins belonging to the
family including the above-mentioned P13 and 4 kinases
have protein kinase activity [Dhand, R., et al., EMBO J.,
13, 522-533 (1994); Stack, J. H. and Emr, S. D., J. Biol.
Chem., 269, 31552-31562 (1994); Hartley, K. 0., et al.,
Cell, 82, 848-856 (1995)].
It was also revealed that a protein belonging
to this family is involved in DNA repair [Hartley, K. 0.,
et al., Cell, 82, 849-856 (1995)] and is a causative gene
of ataxia [Savitsky, K., et al., Science, 268, 1749-1753
(1995)].
It can be anticipated that the human NPIK gene-
encoded protein highly homologous to the family of these
PI kinases is a novel enzyme phosphorylating lipids or
..,., -. ._.. ., . .. :-- =-_-_=..,..----. ..... . .. .. .. .. _. .. . .
._.... .. . .
CA 02458872 2004-03-23
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proteins.
According to this example, the novel human NPIK
gene is provided. The use of said gene makes it possible
to detect the expression of said gene in various tissues
and manufacture the human NPIK protein by the technology
of genetic engineering and, through these, it becomes
possible to study lipid- or protein-phosphrylating
enzymes such as mentioned above, study DNA repairing,
study or diagnose diseases in which these are involved,
for example cancer, and screen out and evaluate drugs for
the treatment or prevention thereof.
(4) Construction of an expression vector for fusion
protein
To subclone the coding region for a human NPIK
gene (GEN-428B12c2), first of all, two primers, Cl and
C2, having the sequences shown below in Table 10 were
formed based on the information on the DNA sequences
obtained above in (1).
Table 10
Primer Nucleotide sequence
Primer Cl 5'-CTCAGATCTATGGGAGATACAGTAGTGGAGC-3'
Primer C2 5'-TCGAGATCTTCACATGATGCCGTTGGTGAG-3'
Both of the primers Cl and C2 have a BglII
site, and primer C2 is an antisense primer.
Using these two primers, cDNA derived from
CA 02458872 2004-03-23
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human fetal brain mRNA was amplified by PCR to provide a
product having a length of about 2500 bases. The
amplified cDNA was precipitated from ethanol and inserted
into pT7BlueT-Vector (product of Novagen) and subcloning
was completed. The entire sequence was determined in the
same manner as above in Examples. As a result, it was
revealed that this gene had polymorphism shown above in
Table 9.
The above cDNA was cleaved by BQlII and
subjected to agarose gel electrophoresis. The cDNA was
then excised from agarose gel and collected using
GENECLEAN II KIT (product of Bio 101). The cDNA was
inserted into pBlueBacHis2B-Vector (product of
Invitrogen) at the BalII cleavage site and subcloning was
completed.
The fusion vector thus obtained had a BglII
cleavage site and was an expression vector for a fusion
protein of the contemplated gene product (about 91 kd)
and 38 amino acids derived from pBlueBacHis2B-Vector and
containing a polyhistidine region and an epitope
recognizing Anti-XpressTM antibody (product of
Invitrogen).
(5) Transfection into insect cell Sf-9
The human NPIK gene was expressed according to
the Baculovirus expression system. Baculovirus is a
CA 02458872 2004-03-23
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cyclic double-stranded insect-pathogenic virus and can
produce large amounts of inclusion bodies named
polyhedrins in the cells of insects. Using Bac-N-B1ueTM
Transfection Kit utilizing this characteristic of
Baculovirus and developed by Invitrogen, the Baculovirus
expression was carried out.
Stated more specifically, 4 pg of pBlueBacHis2B
containing the region of the human NPIK gene and 1 pg of
Bac-N-BlueTM DNA (product of Invitrogen) were co-
transfected into Sf-9 cells in the presence of InsectinTM
liposomes (product of Invitrogen).
Prior to co-transfection, LacZ gene was
incorporated into Bac-N-B1ueTM DNA, so that LacZ would be
expressed only when homologous recombination took place
between the Bac-N-BlueTM DNA and pBlueBacHis2B. Thus
when the co-transfected Sf-9 cells were incubated on agar
medium, the plaques of the virus expressing the
contemplated gene were easily detected as blue plaques.
The blue plaques were excised from each agar
and suspended in 400 ul of medium to disperse the virus
thereon. The suspension was subjected to centrifugation
to give a supernatant containing the virus. Sf-9 cells
were infected with the virus again to increase the titre
and to obtain a large amount of infective virus solution.
(6) Preparation of human NPIK
CA 02458872 2006-09-22
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The expression of the contemplated human NPIK
gene was confirmed three days after infection with the
virus as follows.
Sf-9 cells were collected and washed with PBS.
The cells were boiled with a SDS-PAGE loading buffer for
5 minutes and SDS-PAGE was performed. According to the
TM
western blot technique using Anti-Xpress as an antibody,
the contemplated protein was detected at the position of
its presumed molecular weight. By contrast, in the case
of control cells uninfected with the virus, no band
corresponding to human NPIK was observed in the same
test.
Stated more specifically, three days after the
infection of 15 flasks (175-cm2, FALCON) of semi-
confluent Sf-9 cells, the cells were harvested and washed
with PBS, followed by resuspension in a buffer (20 mM
Tris/HC1 (pH 7.5), 1 mM EDTA and 1 mM DTT). The
suspended cells were lysed by 4 time-sonications for 30
seconds at 4 C with 30 seconds intervals. The sonicated
cells were subjected to centrifugation and the
supernatant was collected. The protein in the
supernatant was immunoprecipitated using an Anti-Xpress
antibody and obtained as a slurry of protein A-Sepharose
beads. The slurry was boiled with a SDS-PAGE loading
buffer for 5 minutes. SDS-PAGE was performed for
CA 02458872 2004-03-23
-84-
identification and quantification of NPIK. The slurry
itself was subjected to the following assaying.
(7) Confirmation of P14 Kinase activity
NPIK was expected to have the activity of
incorporation phosphoric acid at the 4-position of the
inositol ring of phosphatidylinositol (PI), namely, P14
Kinase activity.
P14 Kinase activity of NPIK was assayed
according to the method of Takenawa, et al. (Yamakawa,-A.
and Takenawa, T., J. Biol. Chem., 263, 17555-17560
(1988)) as shown below.
First prepared was a mixture of 10 l of a NPIK
slurry (20 mM Tris/HCl (pH 7.5), 1 mM EDTA, 1 mM DTT and
50t protein A beads), 10 ul of a PI solution (prepared by
drying 5 mg of a PI-containing commercial chloroform
solution in a stream of nitrogen onto a glass tube wall,
adding 1 ml of 20 mM Tris/HC1 (pH 7.5) buffer and forming
micelles by sonication), 10 ul of an applied buffer (210
mM Tris/HC1 (pH 7.5), 5 mM EGTA and 100 mM MgCl2) and 10
l of distilled water. Thereto was added 10 l of an ATP
solution (5 l of 500 pM ATP, 4.9 ul of distilled water
and 0.1 ul of y-32P ATP (6000 Ci/mmol, product of NEN
Co., Ltd.)). The reaction was started at 30 C and
continued for 2, 5, 10 and 20 minutes. The time 10
minutes was set as incubation time because a straight-
--------------- -
CA 02458872 2004-03-23
-85-
line increase was observed around 10 minutes in
incorporation of phosphoric acid into PI in the assaying
process described below.
After completion of the reaction, PI was
fractionated by the solvent extraction method and finally
re-suspended in chloroform. The suspension was developed
by thin layer chromatography (TLC) and the radioactivity
of the reaction product at the PI4P-position was assayed
using an analyzer (trade name: Bio-Image; product of Fuji
Photo Film Co., Ltd.).
Fig. 1 shows the results. Fig. 1 is an
analytical diagram of the results of assaying the
radioactivity based on TLC as mentioned above. The right
lane (2) is the fraction of Sf-9 cell cytoplasm infected
with the NPIK-containing virus, whereas the left lane (1)
is the fraction of uninfected Sf-9 cell cytoplasm.
Also, predetermined amounts of Triton X-100 and
adenosine were added to the above reaction system to
check how such addition would affect the P14 Kinase
activity. The P14 Kinase activity was assayed in the
same manner as above.
Fig. 2 shows the results. The results
confirmed that NPIK had a typical P14 Kinaze activity
accelarated by Triton X-100 and inhibited by adenosine.
~,.:..,.s-------------- ------ --------
CA 02458872 2004-03-23
-86-
Example 10
nel-related protein type 1 (NRP1) gene and nel-related
protein type 2 (NRP2) gene
(1) Cloning and DNA sequencing of NRP1 gene and NRP2
gene
EGF-like repeats have been found in many
membrane proteins and in proteins related to growth
regulation and differentiation. This motif seems to be
involved in protein-protein interactions.
Recently, a gene encoding nel, a novel peptide
containing five EGF-like repeats, was cloned from a chick
embryonic cDNA library [Matsuhashi, S., et al., Dev.
Dynamics, 203, 212-222 (1995)]. This product is
considered to be a transmembrane molecule with its EGF-
like repeats in the extracellular domain. A 4.5 kb
transcript (nel mRNA) is expressed in various tissues at
the embryonic stage and exclusively in brain and retina
after hatching.
Following the procedure of Example 1 (1), cDNA
clones were randomly selected from a human fetal brain
cDNA library and subjected to sequence analysis, followed
by database searching. As a result, two cDNA clones with
significantly high homology to the above-mentioned nel
were found and named GEN-073E07 and GEN-093E05,
respectively.
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Since both clones were lacking in the 5'
portion, 5' RACE was performed in the same manner as in
Example 2 (2) to obtain the entire coding regions.
As for the primers for 5' RACE, primers having
an arbitrary sequence obtained from the cDNA sequences of
the above clones were synthesized while the anchor primer
attached to a commercial kit was used as such.
5' RACE clones obtained from the PCR were
sequenced and the sequences seemingly covering the entire
coding regions of both genes were obtained. These genes
were respectively named nel-related protein type 1 (NRP1)
gene and nel-related protein type 2 (NRP2) gene.
The NRP1 gene contains an open reading frame of
2,430 nucleotides, as shown'under SEQ ID NO:35, the amino
acid sequence deduced therefrom comprises 810 amino acid
residues, as shown under SEQ ID N0:34, and the nucleotide
sequence of the entire cDNA clone of said NRPI gene
comprises 2,977 nucleotides, as shown under SEQ ID N0:36.
On the other hand, the NRP2 gene contains an
open reading frame of 2,448 nucleotides, as shown under
SEQ ID NO:38, the amino acid sequence deduced therefrom
comprises 816 amino acid residues, as shown under SEQ ID
NO:37, and the nucleotide sequence of the entire cDNA
clone of said NRP2 gene comprises 3,198 nucleotides, as
shown under SEQ ID NO:39.
...~:.-=__-_.._..,_______________. - .r.. . . . . .._ :" " '
.=%nrs':;t+r~r{,!'.-
- -:. _.._. _.-_-==._._ _. .. . ..... _..___'__`-.._._.. ... ........ ... - -
.._=-. _ _^_-'_' -__. ._ .....__ _ .._.......y._. . .. .. _ .. . ...__ _....-
".-..-:.- __ _ .e... . _ . . . .
CA 02458872 2004-03-23
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Furthermore, the coding regions were amplified
by RT-PCR to exclude the possibility that either of the
sequences obtained was a chimeric cDNA.
The deduced NRP1 and NRP2 gene products both
showed highly hydrophobic N termini capable of func-
tioning as signal peptides for membrane insertion. As
compared with chick embryonic nel, they both appeared to
have no hydrophobic transmembrane domain. Comparison
among NRP1, NRP2 and nel with respect to the deduced
peptide sequences revealed that NRP2 has 80$ homology on
the amino acid level and is more closely related to nel
than NRP1 having 504 homology. The cysteine residues in
cysteine-rich domains and EGF-like repeats were found
completely conserved.
The most remarkable difference between the NRPs
and the chick protein was that the human homologs lack
the putative transmembrane domain of nel. However, even
in this lacking region, the nucleotide sequences of NRPs
were very similar to that of nel. Furthermore, the two
NRPs each possessed six EGF-like repeats, whereas nel has
only five.
Other unique motifs of nel as reported by
Matsuhashi et al. [Matsuhashi, S., et al., Dev. Dynamics,
203, 212-222 (1995)] were also found in the NRPs at
equivalent positions. Since as mentioned above, it was
CA 02458872 2004-03-23
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shown that the two deduced NRP peptides are not
transmembrane proteins, the NRPs might be secretory
proteins or proteins anchored to membranes as a result of
posttranslational modification.
The present inventors speculate that NRPs might
function as ligands by stimulating other molecules such
as EGF receptors. The present inventors further found
that an extra EGF-like repeat could be encoded in nel
upon frame shifting of the membrane domain region of nel.
When paralleled and compared with NRP2 and nel,
the frame-shifted amino acid sequence showed similarities
over the whole range of NRP2 and of nel, suggesting that
NRP2 might be a human counterpart of nel. In contrast,
NRP1 is considered to be not a human counterpart of nel
but a homologous gene.
(2) Northern blot analysis
Northern blot analysis was carried out as des-
cribed in Example 1 (2). Thus, the entire sequences of
both clones cDNAs were amplified by PCR, the PCR products
were purified and labeled with [32P]-dCTP (random-primed
DNA labeling kit, Boehringer Mannheim) and human normal
tissues were examined for NRP mRNA expression using an
MTN blot with the labeled products as two probes.
Sixteen adult tissues and four human fetal
tissues were examined for the expression pattern of two
CA 02458872 2004-03-23
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NRPs.
As a result of the Northern blot analysis, it
was found that a 3.5 kb transcript of NRP1 was weakly
expressed in fetal and adult brain and kidney. A 3.6 kb
transcript of NRP2 was strongly expressed in adult and
fetal brain alone, with weak expression thereof in fetal
kidney as well.
This suggests that NRPs might play a brain-
specific role, for example as signal molecules for growth
regulation. In addition, these genes might have a
particular function in kidney.
(3) Chromosomal mapping of NRPl gene and NRP2 gene by
FISH
Chromosomal mapping of the NRP1 gene and NRP2
gene was performed by FISH as described in Example 1 (3).
As a result, it was revealed that the
chromosomal locus of the NRP1 gene is localized to
llpl5.1-p15.2 and the chromosomal locus of the NRP2 gene
to 12q13.11-q13.12.
According to the present invention, the novel
human NRP1 gene and NRP2 gene are provided and the use of
said genes makes it possible to detect the expression of
said genes in various tissues and produce the human NRP1
and NRP2 proteins by the technology of genetic
engineering. They can further be used in the study of
--- ---------------- ---------
CA 02458872 2004-03-23
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the brain neurotransmission system, diagnosis of various
diseases related to neurotransmission in the brain, and
the screening and evaluation of drugs for the treatment
and prevention of such diseases. Furthermore, the
possibility is suggested that these EGF domain-containing
NRPs act as growth factors in brain, hence they may be
useful in the diagnosis and treatment of various kinds of
intracerebral tumor and effective in nerve regeneration
in cases of degenerative nervous diseases.
Example 11
GSPT1-related protein (GSPT1-TK) gene
(1) GSPT1-TK gene cloning and DNA sequencing
The human GSPT1 gene is one of the human
homologous genes of the yeast GSTl gene that encodes the
GTP-binding protein essential for the Gi to S phase
transition in the cell cycle. The yeast GST1 gene, first
identified as a protein capable of complementing a
temperature-sensitive gstl (G1-to-S transition) mutant of
Saccharomyces cerevisiae, was isolated from a yeast
genomic library [Kikuchi, Y., Shimatake, H. and Kikuchi,
A., EMBO J., 7, 1175-1182 (1988)] and encoded a protein
with a target site of cAMP-dependent protein kinases and
a GTPase domain.
The human GSPT1 gene was isolated from a KB
cell cDNA library by hybridization using the yeast GST1
CA 02458872 2004-03-23
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gene as a probe [Hoshino, S., Miyazawa, H., Enomoto, T.,
Hanaoka, F., Kikuchi, Y., Kikuchi, A. and Ui, M., EMBO
J., 8, 3807-3814 (1989)]. The deduced protein of said
GSPT1 gene, like yeast GST1, has a GTP-binding domain and
a GTPase activity center, and plays an important role in
cell proliferation.
Furthermore, a breakpoint for chromosome re-
arrangement has been observed in the GSPT1 gene located
in the chromosomal locus 16p13.3 in patients with acute
nonlymphocytic leukemia (ANLL) [Ozawa, K., Murakami, Y.,
Eki, T., Yokoyama, K. Soeda, E., Hoshino, S. Ui, M. and
Hanaoka, F., Somatic Cell and Molecular Genet., 18, 189-
194 (1992)].
cDNA clones were randomly selected from a human
fetal brain cDNA library and subjected to sequence
analysis as described in Example 1 (1) and database
searching was performed and, as a result, a clone having
a 0.3 kb cDNA sequence highly homologous to the above-
mentioned GSPT1 gene was found and named GEN-077A09. The
GEN-077A09 clone seemed to be lacking in the 5' region,
so that 5' RACE was carried out in the same manner as in
Example 2 (2) to obtain the entire coding region.
The primers used for the 5' RACE were P1 and P2
primers respectively having the nucleotide sequences
shown in Table 11 as designed based on the known cDNA
.... .... .......
,._.... . .
-,._.:__...._ ._.....
CA 02458872 2004-03-23
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sequence of the above-mentioned cDNA, and the anchor
primer used was the one attached to the commercial kit.
Thirtyfive cycles of PCR were performed under the
following conditions: 94 C for 45 seconds, 58 C for 45
seconds and 72 C for 2 minutes. Finally, elongation
reaction was carried out at 72 C for 7 minutes.
Table 11
Primer Nucleotide sequence
P1 primer 5'-GATTTGTGCTCAATAATCACTATCTGAA-3'
P2 primer 5'-GGTTACTAGGATCACAAAGTATGAATTCTGGAA-3'
Several of the 5' RACE clones obtained from the
above PCR were sequenced and the base sequence of that
cDNA clone showing overlapping between the 5' RACE clones
and the GEN-077A09 clone was determined to thereby reveal
the sequence regarded as covering the entire coding
region. This was named GSPT1-related protein "GSPT1-TK
gene".
The GSPT1-TK gene was found to contain an open
reading frame of 1,497 nucleotides, as shown under SEQ ID
N0:41. The amino acid sequence deduced therefrom
contained 499 amino acid residues, as shown under SEQ ID
N0:40.
The nucleotide sequence of the whole cDNA clone
of the GSPT1-TK gene was found to comprise 2,057
nucleotides, as shown under SEQ ID N0:42, and the
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molecular weight was calculated at 55,740 daltons.
The first methionine code (ATG) in the open
reading frame had no in-frame termination codon but this
ATG was surrounded by a sequence similar to the Kozak
consensus sequence for translational initiation.
Therefore, it was concluded that this ATG triplet
occurring in positions 144-146 of the relevant sequence
is the initiation codon.
Furthermore, a polyadenylation signal, AATAAA,
was observed 13 nucleotides upstream from the
polyadenylation site.
Human GSPT1-TK contains a glutamic acid rich
region near the N terminus, and 18 of 20 glutamic acid
residues occurring in this region of human GSPT1-TK are
conserved and align perfectly with those of the human
GSPT1 protein. Several regions (G1, G2, G3, G4 and G5)
of GTP-binding proteins that are responsible for guanine
nucleotide binding and hydrolysis were found conserved in
the GSPT1-TK protein just as in the human GSPT1 protein.
Thus, the DNA sequence of human GSPT1-TK was
found 89.4% identical, and the amino acid sequence
deduced therefrom 92.4% identical, with the corresponding
sequence of human GSPT1 which supposedly plays an
important role in the G1 to S phase transition in the
cell cycle. Said amino acid sequence showed 50.8%
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identity with that of yeast GST1.
(2) Northern blot analysis
Northern blot analysis was carried out as des-
cribed in Example 1 (2). Thus, the GEN-077A09 cDNA clone
was amplified by PCR, the PCR product was purified and
labeled with [32P]-dCTP (random-primed DNA labeling kit,
Boehringer Mannheim), and normal human tissues were
examined for the expression of GSPT1-TK mRNA therein
using an MTN blot with the labeled product as a probe.
As a result of the Northern blot analysis, a
2.7 kb major transcript was detected in various tissues.
The level of human GSPT1-TK expression seemed highest in
brain and in testis.
(3) Chromosome mapping of GSPT1-TK gene by FISH
Chromosome mapping of the GSPT1-TK gene was
performed by FISH as described in Example 1 (3).
As a result, it was found that the GSPT1-TK
gene is localized at the chromosomal locus 19p13.3. In
this chromosomal localization site, reciprocal location
has been observed very frequently in cases of acute
lymphocytic leukemia (ALL) and acute myeloid leukemia
(AML). In addition, it is reported that acute non-
lymphocytic leukemia (ANLL) is associated with re-
arrangements involving the human GSPT1 region [Ozawa, K.,
Murakami, Y., Eki, T., Yokoyama, K., Soeda, E., Hoshino,
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S., Ui, M. and Hanaoka, F., Somatic Cell and Molecular
Genet., 18, 189-194 (1992)].
In view of the above, it is suggested that this
gene is the best candidate gene associated with ALL and
AML.
In accordance with the present invention, the
novel human GSPT1-TK gene is provided and the use of said
gene makes it possible to detect the expression of said
gene in various tissues and produce the human GSPT1-TK
protein by the technology of genetic engineering. These
can be used in the studies of cell proliferation, as
mentioned above, and further make it possible to diagnose
various diseases associated with the chromosomal locus of
this gene, for example acute myelocytic leukemia. This
is because translocation of this gene may result in
decomposition of the GSPT1-TK gene and further some or
other fused protein expressed upon said translocation may
cause such diseases.
Furthermore, it is expected that diagnosis and
treatment of said diseases can be made possible by
producing antibodies to such fused protein, revealing the
intracellular localization of said protein and examining
its expression specific to said diseases. Therefore, it
is also expected that the use of the gene of the present
invention,makes it possible to screen out and evaluate
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drugs for the treatment and prevention of said diseases.
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SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT: Tsutomu, FUJIWARA
Takeshi, WATANABE
Masato, HORIE
Toyomasa, KATAGIRI
(ii) TITLE OF INVENTION: HUMAN GENE
(iii) NUMBER OF SEQUENCES: 42
CORRESPONDENCE ADDRESS:
MARKS & CLERK
P.O.BOX 957 STATION B
OTTAWA, ONTARIO
CANADA K1P 5S7
(v) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk
(B) COMPUTER: IBM PC compatible
(C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: PatentIn Release #1.0, Version #1.30
CURRENT APPLICATION DATA
APPLICATION NUMBER: 2458872
FILING DATE: 19-Mar-1997
CLASSIFICATION:
PRIOR APPLICATION DATA
APPLICATION NUMBER:
FILING DATE:
CLASSSIFICATION:
PATENT AGENT INFORMATION
NAME: MARKS & CLERK
REFERENCE NUMBER: 16487-9-DIV
(2) INFORMATION FOR SEQ ID NO:1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 122 amino acids
(B) TYPE: amino acid
... CA 02458872 2004-03-23 - - -
-99-
(ii) MOLECULE TYPE: DNA(cDNA)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:
ATGGAGTI'GG GGAAGATGG CAGTGZCTAT AAGAOCATTT TOOIGACAAG CCAGGACAAG 60
GCTCCAAGrG TCAT~',AGZCG TGI'C7C;TTAAG AAAAACAATC GIGACPCI'GC AGPGOCPTCA 120
GAG'rATGAGC TGGTACAOCT GCTAOC',AGGG GAGOC'~A~f'~C TGACTATOOC AOOCTCOOCT 180
AATGTAT'PCT ACOCCATGGA TGC'aAGCTI'CA CAIOC',AT'I"I'OC TOC7lG~OC~OCA GOGGOCAAGG
240
ZCX.'iC.TAGTG C.TACACC70G aGI'CACCAGT GGOOOGTG7.'G CX,TC.AOGAAC
TOCPOOGA+C~P300
GAOGC'~GGAG GGOC~CTOf:'1T TO(X'.AGX"ATC AAGGOCACAG GGAOC'aAP,GAT TGCAC7GGGC'A
360
CTGriC 366
(2) INFORMATION FOR SEQ ID NO:3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 842 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-501D08
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 28..393
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:
CCCACGAGOC GTATCA4CCG AG'POC'AG ATG GAG TTG GGG GAA GAT GGC AGT 51
Met Glu Leu Gly Glu Asp Gly Ser
1 5
GTC TAT AAG AGC ATT TTG GTG ACA AGC CAG GAC AAG GCT CCA AGT GTC 99
CA 02458872 2004-03-23
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Val Tyr Lys Ser Ile Lau Val Thr Ser Gln Asp Lys Ala Prro Ser Val
15 20
ATC AGT UG`I' GTC CIT AAG AAA AAC AAT aGT GAC TCT GCA GTG GCT TCA 147
Ile Ser Arg Val Lau Lys Lys Asn Asn Arg Asp Ser Ala Val Ala Ser
25 30 35 40
GAG TAT GAG CTG GTA CAG CTG CTA OC'A GGGG GAG OGA GAG CIG ACT ATC 195
Glu Tyr Glu L8u Val Gln Leu Leu Pro Gly Glu Arg Glu Leu Thr Ile
45 50 55
aCA GOC TrJG GC.T AAT GTA TTC TAC QOC ATG GAT GGA GCT TCA CAC GAT 243
Pro Ala Ser A.la. Asn Val Phe Tyr Pro Met Asp Giy Ala Ser His Asp
60 65 70
TTC CI'C CTG QOG CAG aGG OGA AOG TIJC TCT ACT GCT ACA aCT GOC GTC 291
Phe Leu Leu Asg Gln Arg Arg Arg Ser Ser Thr Ala Thr Pro Gly Val
75 80 85
AOC AGT GGC CCG TCT GOC TCA GGA ACT OCT aCG AGr GAG GGA GGA GGG 339
Thr Ser Gly Pro Ser Ala Ser Gly Tbr Pro Pro Ser Giu Gly Gly Gly
90 95 100
GGC. TOC TTT CCC AOG ATC AAG GfJC ACA GGG AGG AAG ATT GCA aGG GCA 387
Gly Ser Phe Pro Arg Ile Lys Ala Thr Gly Arg Lys Ile Ala Arg Ala
105 110 115 120
CIG Tt^C TGAGGAGGAA OCOCI"PI'1'T TTTACAGAAGT CAT~'IGTTC ATACCAGATG 443
Leu Phe
TGOGG'rAOCCA TaCIC`aAATGG TGGCAATTAT ATCACATTGA GACAGAAATT CAGkAAGGGA 503
GOCAGCCAiOC C'1GGGGCAGT GAAGTGOC'AC TGGITPAUCA GACAGCTGAG AAATCX',AOOC 563
CTGICGGAAC TGGI'GI'C,TPA TAAOC'.AA~siT OGATAUCIGT GTATAGGTIG CCACCTTCC'A 623
TGAGTGCAGC ACACAOGTAG TOCI'GGAAAA AOGCAZCAGP TPCZGF-T'IC,`I' TGGCC'ATATC 683
CTAACATGOA AOOGCC'.A11GC AAAGGCiT('A AOGCPCIY''AG CCCC',AGGGC:A GAGGOC'~ATG
743
GCAAAATGTA GGPCC`I'GGCA GGAGCTCT'IC TTCCCACTCP aGAGMITPCT ATCACPGTGA 803
CAAICACTAAG ATAATAAAOC AAAACACTAC CIC'7AATTCT 842
(2) INFORMATION FOR SEQ ID NO:4:
( i ) SEQUENCE QHP-RACTERISTICS:
(A) LENGPH : 193 amino acids
( B ) TYPE: amino acid
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( D ) TOPOL4G'Y: linear
(ii) MDLBCULE TYPE: protein
( xi. ) SEQUIINCE DESCRIPTION: SEQ ID PK): 4:
Met Glu Leu Glu Leu Tyr Gly Val Asp Asp Lys Phe Tyr Ser Lys Leu
1 5 10 15
Asp Gln Glu Asp Ala Leu Leu Gly Ser Tyr Pro Val Asp Asp Gly Cys
20 25 30
Arg Ile His Val Ile Asp His Ser Gly Ala Arg Leu Gly Glu Tyr Glu
35 40 45
Asp Val Ser Arg Val Glu Lys Tyr Thr Ile Ser Gln Glu Ala Tyr Asp
50 55 60
Gin Arg Gln Asp Thr Val Arrg Ser Phe Leu Lys Arg Ser Lys Leu Gly
65 70 75 80
Arg Tyr Asn Giu Glu Glu Arg Ala Gln Gln Glu Ala Glu Ala Ala Gln
85 90 95
Arg Leu Ala Glu Glu Lys Ala Gin Ala Ser Ser Ile Pro Vai Gly Ser
100 105 110
Arg Cys Glu Val Arg Ala Ala Gly Gln Ser Pro Arg Arg Gly Thr Val
115 120 125
Met Tyr Vai Gly Leu Thr Asp Phe Lys Pro Gly Tyr Trp Ile Gly Vai
130 135 140
Arg Tyr Asp Glu Pro Ip-u Gly Lys Asn Asp Gly Ser Val Asn Giy Lys
145 150 155 160
Arg Tyr Phe Glu Cys Gln Ala Lys Tyr Giy Ala Phe Val Lys Pro Ala
165 170 175
Val Val Thr Val Gly Asp Phe Pro Glu Glu Asp Tyr Gly LSu Asp Glu
180 185 190
Ile
(2) INFORMATION FOR SEQ ID N0:5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 579 base pairs
CA 02458872 2004-03-23
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(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(cDNA)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:
ATGC'aAAiCI'GG AGCPGTATGG AGrPIGACGAC AAGTPCTACA GCAAGCPOGA TCAAGAGGAT 60
GOOLZ~OCICAG GCPaGTAOOC TGTAGATW GGC.'TOOOpC'A TC7C'AiOGPCAT TGACCACAG'r 120
GO~JG00f700C TTOQIG~PA ZG'A0GGAOOIG TOOCJOQOIC~G AiGAAGTAC~C GATCTCACAA 180
GAAGCCPAUG ACCAGAGGC'A AGACAOGGIC (70CIC'1TPOC TGAAGCGC',AG CAAOCTCGGC 240
CGGTAICAAOC' AGGAGGAGOG GGC'IPCAGCAG GAGGOOG.AOG CCGOOCAOOG CCiG000f"aAG 300
GAGAAGGOUC AOGOCAOCIC CATOOOOG'IG GOC'19OCOOC.'T GTGAQGTOOG GGOOOCGGC'aA 360
CAARti700C,'TC GOOGGGOCAC OGICATGPAT GTAOGt'CPCA CAGP-TPt'CAA GGC7POGCPAC 420
TGGA'PInGIG TUOC3CPATGA TGAGOC',AIC,ZG GGGAAAAATG ATGGCAGTGr GAATG3C'=AAA 480
aGC'PACT'I'OG AATGO('.AOOC CAAGTATG3C GOC'iTl'C~PCA AGOC,AGCAGT aGIC'~AIOGGIPG
540
f,GGGGF-CPI'CC OGGAGGAGC', CTACGGGPTG GAL7GGAGATA 579
(2) INFORMATION FOR SEQ ID NO:6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1015 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-080G01
(ix) FEATURE:
(A) NAME/KEY: CDS
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(B) LOCATION: 274..852
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:
TGATZGG'7CA GGCACGC"aAOC AGGAGGOOGG CTGIATAGOOC AGCAGCAGCA GCGOOG70COG 60
QOGCt~G00C'~1 GOOOG'1C~lGA GOOOOC'1C1("aA QOOOGCPGCA GOCAT00GM GGCGOGOCAA 120
GATGGAGGTG AOGaGGG'I'GT CGGCACCAOG GTGA00 1TT TCATCAGCAG CTCOCICAGC 180
AOCT'l~OOOC'P COGAL''aAAGOG ATACAIG100GC AGOCPCAOCA TOOCt~GA[~IT CAAGTGTAAA
240
C'lGGAiGPTGC TGGTOGGC.AG aC)CTGC= Z'i0C ATG GAA CIG GAG CIPG TAT GGA 294
Met Glu Lau Glu Leu Tyr Gly
1 5
G'1T GAC GAC AAG TTC TAC AOC AAG CTG GAT CAA GAG GAT GOG CTC GRG 342
Val Asp Asp Lys Phe Tyr Ser Lys Lau Asp Gin Glu Asp Ala Lau Leu
15 20
GGC TCC TAC aCI' GTA GAT GAC GOC TOC QGC ATC CAC GZC ATT GAC CAC 390
Gly Ser Tyr Pro Val Asp Asp Gly Cys Arg Ile His Val Ile Asp His
25 30 35
AGT GGC GCC CGC CiT GGT GAG TAT GAG GAC GTG TCC CGG GTG GAG AAG 438
Ser Gly Ala Arg Leu Gly Glu Tyr Glu Asp Val Ser Arg Val Glu Lys
40 45 50 55
TAC AIOG ATC TCA CAA GAA GCC TAC GAC CAG AOG CAA GAC ACG GTC COC 486
Tyr Thr Ile Ser Gln Glu Ala Tyr Asp Gin Arg Gln Asp Thr Val Arg
60 65 70
TCT TTC CTG AAG CGC AOC AAG CTC GOC CGG TAC AAC GAG GAG GAG CGG 534
Ser Phe Leu Lys Arg Ser Lys Leu Gly Arg Tyr Asn Glu Glu Glu Arg
75 80 85
GCT CAG CAG GAG GOC GAG GOC GOC CAG CGC C'IG GOC GAG GAG AAG GOC 582,
Ala Gin Gln Glu Ala Glu Ala Ala Gln Arg Leu Ala Glu Glu Lys Ala
90 95 100
CAG GOC AOC TCC ATC COC GTG GOC AGC QOC TIaT GAG G'iG COG GOG GflG 630
Gln Ala Ser Ser Ile Pro Val Gly Ser Arg Cys Glu Val Arg Ala Ala
105 110 115
GGF- CAA TCC CCT CGC CGG GGC ACC GTC ATG TAT GTA GGT CTC ACA GAT 678
Gly Gln Ser Pro Arg Ar+g Gly Thr Val. Met Tyr Val Gly Leu Thr Asp
120 125 130 135
TTC AAG CCT GGC TAC TGG ATT GGT GTC CGC TAT GAT GAG OCA CTG GGG 726
CA 02458872 2004-03-23
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Phe Lys Pro Gly Tyr Trp Ile Gly Val Arg Tyr Asp Glu Pro ILeu Gly
140 145 150
AAA AAT GAT GGC AGT GTG AAT GtflG AAA CGC TAC TTC GAA TGC CAG GOC 774
Lys Asn Asp Gly Ser Val Asn Gly Lys Arg Tyr Phe Glu Cys Gln Ala
155 160 165
AAG TAT G{flC GCC TTT Gl'C AAG C7CA GCA GTC GTG ACG GIG GGG GAC TTC 822
Lys Tyr Gly Ala Phe Val Lys Pro Ala Val Val Thr Val Gly Asp Phe
170 175 180
DOG GAG GAG GAC TAC GOG TTG GAC GAG ATA T('~"1lQCTAA GGAATPCCW 872
Pro Glu Glu Asp Tyr Gly Leu Asp Glu Ile
185 190
OCACTGAM C70C(.'1~OC71C~P G1C~'TC~OOCAT OOOC'PT1R~C 932
TOCTTCAOCT aCTAG~CICAG
TO(."1C'aA000C ATTTTAAT'PP TATTCATTTT '1"1~OC1Ti'OOC ATTGATTTTP GA("aAC1CATG
992
CATTAAATTC ACTAGAAAOC CAG 1015
(2) INFOBMTION FX)Z SEQ ID N0: 7:
( i ) SEXYJE24CE CHARACTERISTICS:
( A ) IEVGT'H: 128 aniro acids
( B ) TYPE : amino a.id
( D ) TOPOLOGY: linear
(ii) MOLEQJLE TYPE: protein
( xi ) SEQUENCE DES(RIPTION: SP7Q ID N0: 7:
Met Thr Glu Ala Asp Val Asn Pro Lys Ala Tyr Pro Leu Ala Asp Ala
1 5 10 15
His Leu Thr Lys Lys Leu Leu Asp Leu Val Gln Gln Ser Cys Asn Tyr
20 25 30
Lys Gln Leu Arg Lys Gly Ala Asn Glu Ala Tthr Lys Thr Leu Asn Arg
35 40 45
Gly Ile Ser Glu Phe Ile Val Met Ala Ala Asp Ala Glu Pro Leu Glu
50 55 60
Ile Ile Leu His Leu Pro Leu Leu Cys Glu Asp Lys Asn Val Pro Tyr
65 70 75 80
Val Phe Val Arg Ser Lys Gln Ala Leu Gly Arg Ala Cys Gly Val Ser
CA 02458872 2004-03-23
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85 90 95
Arg Pro Val Ile Ala Cys Ser Val Thr Ile Lys Glu Gly Ser Gin Leu
100 105 110
Lys Gln Gln Ile Gln Ser Ile Gln Gln Ser Ile Glu Arg Leu Leu Val
115 120 125
(2) INFORMATION FOR SEQ ID NO:8:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 384 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:
ATGACTGAGG CTGATGIC'aAA TOCAAAGGOC TATOOOGR'Ir' OOCaAZGO0C.A OCICAC7CAAG 60
AAIX,'I'ACPGG ACCIC7GTTCA GCAGICATGT AACTATAAQC AOL'PInGC'AA AOGAGOCAAT 120
GAOG(7CAOCA AAACOCICAA CAGGGGCATC TCTGAGjTTCA TOQI'GATGGC TGCCAGACGOC 180
GAGCCAC:TOG AGATCATTCT GCAIOCI~OCX7G CTOCTGIv1G AAGACAAGAA TGPGOOCPAC 240
G1GTrT(:TGC GC'IC7CAAGCA GGOOC,ZC~~OOG AGl4GOCIGPG G(~GPCTOCAG GOCiCTCATC 300
GOC'I~CTC'AC('ATCAA A~'aAAGOC1POGG CAGiC'IGAAAC AGCAC'aATOCA ATOCAT'ICAG 360
CAG9OCATTG AAAOGCPGTP AGPC 384
(2) INFORMATION FOR SEQ ID N0:9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1493 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
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(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-025F07
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 95..478
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:
ATCOGTO'i~OC TTGOaQrOC.T GGGCAGCAGA COfOPCCAAW C7GACACGOG'P GG'TATO(.'TC(' 60
OOGI~GTOOOG CAAGAGAGTA CavrAcwA OOQP A7G AGT GAG GCT GAT GTG 112
Met Thr Glu Ala Asp Val
1 5
AAT CCA AP-G GCC TAT CCC C3'T GOC GAT GOC CAC C'iC ACC A14G AAG CTA 160
Asn Pro Lys Ala Tyr Pro IQU Ala Asp Ala His Leu Thr Lys Lys Leu
15 20
CTG GAC CTC GTT CAG CAG TCA TGT AAC TAT AAG CAG CTT COG AAA GGA 208
Lau Asp Leu Val Gln Gln Ser Cys Asn Tyr Lys Gln Leu Ar+g Lys Gly
25 30 35
GOC AAT GAG GOC AOC AAA AUC CTC AAC AGG GGC ATC TCT GAG T'PC ATC 256
Ala Asn Glu Ala Thr Lys Thr Leu Asn Arg Gly Ile Ser Glu Prie Ile
40 45 50
GTG ATG GCT GCA GAC GCC GAG CCA CTG GAG ATC ATT CTG CAC CTG COG 304
Val Met Ala Ala Asp Ala Glu Pro Leu Glu Ile Ile Leu His Leu Pro
55 60 65 70
CTG CTG TGT GAA GAC AAG AAT GTG COC TAC GTG TTT GPG COC TOC AAG 352
Leu Leu Cys Glu Asp Lys Asn Val Pro Tyr Val Phe Val Arg Ser Lys
75 80 85
CAG GOC CTG GGG AGA GOC TGT GGG GTC TCC AGG CCT GTC ATC GCC TGT 400
Gin Ala Leu Gly Azg Ala Cys Gly Val Ser Arg Pro Val Ile Ala Cys
90 95 100
TCT GTC ACC ATC AAA GAA GGC TaG CAG CTG AAA CAG CAG ATC CAA TCC 448
Ser Val Thr Ile Lys Glu Gly Ser Gln Leu Lys Gln Gln Ile Gln Ser
105 110 115
ATT CAG CAG TCC ATT GAA AGG CTC TTA GTC TAAACJC'IWG GCCPCIGCXA 498
Ile Gln Gln Ser Ile Glu Arg Leu Lau Val
120 125
CG'I'GC'I'OCCT GOCAGCTTCC COOCTGAGGT 'PG'IGPA'ICAT ATTA'IC'IGIG TTAGCATGTA 558
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GTATTTTOAG CTAC'IC,'ZCPA TPGITATAAA ATGTPuGTACT AAATCTGGIT 1. C1C~C',ATTTT 618
IUTGTTGM T'ITCPGiT TTACAOGM GCTATOOOOC TTOLTTTOGT OOCPOOCPCr 678
GOCATOCt*PC ATOCZ'PITAT OCIPOOCITi'P TOGAACAAGP GPI'CAGAGCA GAC'.AGAAGCA 738
GGGIGGIC'~GC AOOG'1'PGAAA GGC'AC'aAAAGA GOCAGGAiGAA AGCPGATGGA GOCAIDGACAG 798
AGAZCLGGTP OCApCTITCA GCICATAGCT T0.71WIVIG TGOGGC~GTGT GGPOCaAATTA 858
AACIAGCATTC ATTGTGZGIC IX,TG'i'GOCTC' GCACACAGAA TCATTCATAC GTGTICAAG'P 918
GATCAAOGGG TTPCAZTI"GC TG'PICXXOOC"aA TTAGGTATCA TT'iGGOGAGG AAGCATGTGT 978
TCTGT('aAOGT TGTTCX~OC,TA TGPOC'AA[3IG TOGI'lTACTA AZWAOOOC.T GCTGTIRuCT 1038
TITOGTAATG TGATG`ITC&T G2R~CI~OOOOC TAOOCACAAiC CATGOOCTIG AGGGTAOCAG 1098
GGCAGCAOC'.A TAOCAAAGAG AZG1C +C'IGCA GGACTCOGGA GOC'AOOCI'GG G'TGOGiC'~AIDC
1158
CA.TGGGGCAG 7'1GAOGTOGG TCTIGAAAGA Gt'CGOGAGTG ACAPiGCZCAG AGAG(',ATGAA 1218
CIGAZGCPGf' CATGAAGGAT TGCAOGAAGA TCATGGAGAC CTGOGCPGGTA G.'I'G'PAACAG 1278
AIGATOGI+GG=P- G'IPOf',AAOGAA ACAOOCTGZC TC"IGGIGAAT GOGACP'ITCP
Tl'GGJ.'GGAICA 1338
CZTGGC'AOCA'GCI'CI'GAGAG C707rT0000T GPC~'IYJCTGOC ACCAZGTGOG TCAIGATGTAC 1398
TC7'CPGICAC ATGAOGGAGAU TGCPAGPI"CA TG'I'GI'TCTOC AZRCTI'GIC'~A GCATOCTAAT
1458
AAATCPGI'TC CATTTZGAAA AAAAAAAAAA AAAAA . 1493
(2) INFOEMTION FUEt SDQ ID NO:10:
(1) SnQUIIVr'E CHARACTEPJWICS:
(A) LIIJGTfi: 711 amin4 acids
( B ) TYPE : amiryo acid
( D ) TOPOLOGY: linear
( ii ) MOLEC'1RE TYPE: protein
( xi ) SE7QUQICE DESCRIPTICN: SSQ ID N0:10:
Met Pro Ala Asp Val Asn Leu Ser Gln Lys Pro Gin Val. Leu Gly Pro
1 5 10 15
Glu Lys Gln Asp Gly Ser Cys Glu Ala Ser Val Ser Phe Glu Asp Val
20 25 . 30
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Thr Val Asp Phe Ser Arg Glu Glu Trp Gln Gln LSu Asp Pro Ala Gln
35 40 45
Arg Cys Leu Tyr Arg Asp Val Met leu Glu Leu Tyr Ser His leu Phe
50 55 60
Ala Val Gly Tyr His Ile Pro Asn Pro Glu Val Ile Phe Arg Met Leu
65 70 75 80
Lys Glu Lys Glu Pro Axg Val Glu Glu Ala Glu Val Ser His Gln Arg
85 90 95
Cys Gln Glu Arg Glu Phe Gly leu Glu Ile Pro Gln Lys Glu Ile Ser
100 105 110
Lys Lys Ala Ser Phe Gln Lys Asp Met Val Gly Glu Phe Thr Arg Asp
115 120 125
Gly Ser Trp Cys Ser Ile Leu Glu Glu Leu Arg Lau Asp Ala Asp. Arg
130 135 140
Thr Lys Lys Asp Glu Gln Asn Gln Ile Gln Pro Met Ser His Ser Ala
145 150 155 160
Phe Phe Asn Lys Lys Thr Leu Asn Thr Glu Ser Asn Cys Glu Tyr Lys
165 170 175
Asp Pro Gly Lys Met Ile Arg Thr Arg Pro His Leu Ala Ser Ser Gin
180 185 190
Lys Gin Pro Gln Lys Cys Cys leu Phe Thr Glu Ser Leu Lys Leu Asn
195 200 205
L2u Glu Val Asn Gly Gln Asn Glu Ser Asn Asp Thr Glu Gin Leu Asp
210 215 220
Asp Val Val Gly Ser Gly Gln Lau Phe Ser His Ser Ser Ser Asp Ala
225 230 235 240
Cys Ser Lys Asn Ile His Thr Gly Glu Thr Pthe Cys Lys Gly Asn Gln
245 250 255
Cys Arg Lys Val Cys Gly His Lys Gln Ser Lau Lys Gln His Gln Ile
260 265 270
His Thr Gln Lys Lys Pro Asp Gly Cys Ser Glu Cys Gly Gly Ser Phe
275 280 285
Thr Gln Lys Ser His Leu Phe Ala Gln Gln Arg Ile His Ser Val Gly
290 295 300
CA 02458872 2004-03-23
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Asn Lau His Glu Cys Gly Lys Cys Gly Lys Ala Phe Met Pro Gln Leu
305 310 315 320
Lys Loeu Ser Val Tyr L$u Thr Asp His Thr Gly Asp Ile Pro Cys Ile
325 330 335
Cys Lys Glu Cys Gly Lys Val Phe Ile Gln Arg Ser Glu Lau Leu Thr
340 345 350
His Gln Lys Thr His Thr Arg Lys Lys Pro Tyr Lys Cys His Asp Cys
355 360 365
Gly Lys Ala Phe Phe Gln Met Lsu Ser Leu Phe Arg His Gln Arg Thr
370 375 380
His Ser Arg Glu Lys Lssu Tyr Glu Cys Ser Glu Cys Gly Lys Gly Phe
385 390 395 400
Ser Gln Asn Ser Thr Leu Ile Ile His Gin Lys Ile His Thr Gly Glu
405 410 415
Arg Gln Tyr Ala Cys Ser Glu Cys Gly Lys Ala Phe Thr Gin Lys Ser
420 425 430
Thr Leu Ser Leu His Gln Arg Ile His Ser,Gly Gln Lys Ser Tyr Val
435 440 445
Cys Ile Glu Cys Gly Gln Ala Phe Ile Gln Lys Ala His Leu Ile Val
450 455 460
His Gln Arg Ser His Tlvc Gly Glu Lys Pro Tyr Gln Cys His Asn Cys
465 470 475 480
Gly Lys Ser Phe Ile Ser Lys Ser Gln Lau Asp Ile His His Arg Ile
485 490 495
His Thr Giy Glu Lys Pro Tyr Glu Cys Ser Asp Cys Gly Lys Thr Phe
500 505 510
Thr Gln Lys Ser His Leu Asn Ile His Gln Lys Ile His Thr Giy Glu
515 520 525
Arg His His Val Cys Ser Glu Cys Gly Lys Ala Phe Asn Gln Lys Ser
530 535 540
Ile Leu Ser Met His Gln Arg Ile His Thr Gly Glu Lys Pro Tyr Lys
545 550 555 560
Cys Ser Glu Cys Gly Lys Ala Phe Thr Ser Lys Ser Gln Phe Lys Glu
565 570 575
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His Gin Arg Ile His Thr Gly Glu Lys Pro ZYir Val Cys Thr Glu Cys
580 585 590
Gly Lys Ala Phe Asn Gly Arg Ser Asn Phe His Lys His Gln Ile Thr
595 600 605
His Thr Arg Glu Arg Pro Phe Val Cys Tyr Lys Cys Gly Lys Ala Phe
610 615 ' 620
Val Gin Lys Ser Glu Leu Ile Thr His Gin Arg Thr His Met Gly Glu
625 630 635 640
Lys Pro TyrGlu Cys Leu Asp Cys Gly Lys Ser Phe Ser Lys Lys Pro
645 650 655
Gin Leu Lys Val His Gln Arg Ile His Thr Gly Glu Arg Pro Tyr Val
660 665 670
Cys Ser Glu Cys Gly Lys Ala Phe Asn Asn Arg Ser Asn Phe Asn Lys
675 680 685
His Gln Tnr Tthr His Thr Arg Asp Lys Ser Tyr Lys Cys Ser Tyr Ser
690 695 700
Val Lys Giy Phe T'hr Lys Gln
705 710
(2)' INFORMATION FOR SEQ ID NO:11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2133 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:
ATGOC.'ZbC1G ATGPGAATTT ATCO(',11GAAG CCPC,AOC'IYJC TGGGPOCAGA GAAGCAGGP-T 60
GGATCI"I'GOG AGGCA'ICAGP GTCA7TrGAG GAIOOIC'~AOCG TGGAC'PPCAG CAGGGAG("',~AG
120
TGGC'AGCAAC ZGGACCCTGC CC,A['~'aATGC CTGTAQOGGG ATGIGATGCT GGAGCZCTAT 180
AG(JCATCPCr TCGCAGTOOG GTA'1C'Af' ATT OOCp-ACCCAG AGGTCATGTT CAGAATCGC.'TA 240
AAAGAAAAGG AGOCCCGPGT GGAGGAGGCT GAAGTCPCAC ATCAGAC'G'IG TCAAGAAAGG 300
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GAGTTI'OGOC T'iC'~AAATOOC ACAAAAQCaAG ATTTCrAAGA AAOCI'PCATT TCAAAAOGAT 360
ATaGTAOGIG AGT:rCACAAG AGATGGTTCA TC~~'QTTOCA TTTTAGAAGA ACI'GAOOCIG 420
GA7JaJIY'~iOC (OC',AL'AAAIGAA AGATGAGCAA AATC'AAATTC AAIOOC'ATC',AG
T'CACAG71i0CT 480
TT+CTTCAACA AOAAAACATT GAACACAI~'aAA AO('.AATZGPG AATATAAOGA OOC'POOGAAA 540
ATGAT'iCGCA C7C"~OOOf'A OCTi'C~CTTCT TCIACAC'~AAC AACCPG3GAA ATGlTOC,ZTA 600
TTTACAGAAA GPrTGAAOCT GAAC(.'TAGAA GTGAACOOPC AGAATGAAAO CAATGAC',ACA 660
GAAL:AGC.'TI'G ATGACOI'M TOOGT(,TGG'P CAGCTATTCA DOCATAOCTC TTCTGP~TOOC 720
TOC',A CAAd''aA ATATTCATAC AOGAGAGACA TTTTGCAAAG GTAACC'AQ1G TAGAAAAGI'C 780
ZCsT000C`ATA AAICAIGt'CALT ('AAQCAAC'AT CAAATTCATA C1'"CAGAAGAA AOCAGATOGA 840
Tt~t'ICTG~AAT OIC~OOOOC,Af CTTCA00('.AG AAGTCA('.AiOC T+C?I'1'IC~OOCA
ACAGAf'AATT 900
CATAGrGPAG GAAAL7CTOCA TGAATG'tGGC AAATGl'OGAA AAOOC.'TTCAT GOCACAAC'PA 960
AAACTCAGI'G TATA'I'C,"TGAIC AGATCATACA GGI'C'~ATATAC OCiC~TATATG CAAOGAATGT
1020
OOC'~AAGGT(.'T TTATTCAGAG ATC'.AGFIATTG CTTAOOCAOC AGAAAACACA CAL'TAGAAAG 1080
AAGOOL`PATA AATG'OCATGA CI'G'i'dGAAAA OCPrIRTOC AlGATGTTATC TCTCTTCWA 1140
CATCAGAiGAA CICACAGPAG AC'aAAAAACTC TATOAA7GCA GTGAATGl'OG CAAA[OaCTPC 1200
TOOCAAAACT CAACO(71C'AT TATACAZCAG AAAATTCATA CIGGIGAGAG ACAG'PATOCA 1260
TGCAGrGAAT GPOOC',AAAOC CiTPAUOf'.AG AAiG'I'CAACAC TCAOCPPOCA C7CAGAGAATC 1320
CAICZCAAOOC AGAAIGTC7CTA T(~i'GTC~PATC GAATOOG= AIOGOCTWAT aCAGAA00CA 1380
CACCTGATTG TCCATCAAAG AAGOCACACA GGAGAAAAAC CITATCAGIG GCACAACPOT 1440
GOGAAATCGT TCATI"IC7C'.AA GTCACAGCTT GATATACATC ATOGAATTCA TACAOOGif'aAG 1500
AAAUCTPATG AAT~'1AGiGA C'I'GPOGAAAA AOGTIC:AOOC AAAAQTC'ACA CCl'GAATATA 1560
CAQCA~C'~AAAA TTCATAICIGG AGAAAGACAC CATGTATOCA GTC,AATG170C' GAAAOOCITC 1620
AAOCAGAAG'7.' CAATAC't'CAG CATGCATCAG AGAATTCACA QOOC'~GAGAA GOCZTAiCAAA 1680
TGOCAG'iGAAT GTGGGAAAOC CTTC'ACTTGT AAGI'C,`TCAAT TY'.AAAGAOC'A TCA00('aAATT
1740
C1suCACUGOGTG AGAAAOOCPA TGPGTOCACT GAATG'TGGC''A AOGOCITCAA OOOCAOGPCA 1800
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AATTTOC'ATA AACATCAAAT AAICICACACT AGAGAGAGOC CTT'PTGn`IG TTA(.AAATGT 1860
GOGAACaOGTT TTGICJCAIGAA ATCAIGAGr'IG ATTAOCCATC AAAGAACTCA CA'IGGGAGAG 1920
AAAOOC,TATG AATaCC'=TTGA CTQPOGGAAA TOG'iRCAGrA AGAAAOCõACA'ACPCAAOGZG 1980
CAT('.AGOC'AA TTCACAfJGOG AC'õAAA(`aAC7CT TAZGTv1v1T G'IGAATGZW AAAGGUCT'II'C
2040
AACAACAGGT CAAACITCIAA TAAACAUC'AA ACAAC,'TCATA OC=AG19GAC'.AA ATCTTACAAA 2100
TGCAG7ITATT CTGTIGAAAIOG CTTTACCAP-iG CAA 2133
(2) INFORMATION FOR SEQ ID N0:12:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3754 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-076C09
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 346..2478
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:
GCTAAGOC.'TA TOPOIX.`ITAC TOGAOOC'IGA AGTGATiGG(` AATATTAO('.A CSIGt~C~*ITC 60
TGTAGGGTCA OGAAOOGOOG aCT00C.'TTTG GGGGAGTGAT GAOGOGC,'ZRC' TTGGO GI'GG 120
GGGIGOC~IGA TAAAOOGATT TC.'TOCGCiGA AGACGACGCT GI.'GAOGCI'TC TG('.AGAACOC 180
C3C,AGGI'CAGG OCAC'ATCAZT C',AG1GC'IGC.AIG GAZCTCrCTT CATAGOOC'1UG TACGACTCTC
240
CGCCGI'G1CC CTGGPIGGAA AATOCAAAIC'A CCTATOC'AGC T'IC'ZGGC'I'OC IGG("õAAAAGT
300
GGAGrTGPCA OCAAGAGA(',A aCGAGAGTAG AAGCCCAOAlG ZGC;AG ATG CCT GCT 354
Met Pro Ala
CA 02458872 2004-03-23
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1
GAT GTG AAT TTA TC:C C'AG RAG OC.'T CAG Gl'C CPG =OCA GAG AAG CAG 402
Asp Val Asn Leu Ser Gln Lys Pro Gln Val Leu Gly Pro Glu Lys Gin
10 15
GAT GGA TCT TGC GAG GCA TCA GTG TCA TTT GAG GAC GTG AOC GIG GAC 450
Asp Gly Ser Cys Glu Ala Ser Val Ser Phe Glu Asp Val Thr Val Asp
20 25 30 35
TTC AOC 'AOG GAG GAG TGG CAG CAA CTG GAC aGT GOC CAG AGA TGC CPG 498
Phe Ser Arg Glu Glu Tip Gln Gln Leu Asp Pro Ala Gin Arg Cys Leu
40 45 50
TAC OGG GAT GTG ATG CTG GAG CTC TAT AOC CAT CTC TTC GCA GTG GOG 546
.Tyr Arg Asp Val Met Leu Glu Lau Tyr Ser His Leu Phe Ala Val Gly
55 60 65
TAT CAC ATT (XJC AAC OCA GAG GTC ATC TTC AGA ATG CTA AAA GAA AAG 594.
Tyr His Ile Pro Asn Pro Glu Val Ile Phe Arg Met Leu Lys Glu Lys
70 75 80
GAG OOG OGT GTG GAG GAG GCT GAA GTC TCA CAT CAG AGG TGT CAA GM 642
Glu Pro Arg Val Glu Glu Ala Glu Val Ser His Gin Arg GyS G3.n Glu
85 90 95
AGG GAG TTT GGG C1T GAA ATC OCA CAA AA[", GAG ATT TL'T AAG AAA GCT 690
Arg Glu Phe Gly Leu Glu Ile Pro Gln Lys Glu Ile Ser Lys Lys Ala
100 105 110 115
TGA TTT CAA AAG GAT ATG GTA GQT GAG Ti+C ACA AGA GAT GQP TGA TGG 738
Ser Phe Gln Lys.Asp Met Val Gly Glu Phe Thr Arg Asp Gly Ser Trp
120 125 130
TCT TOC ATT TTA GAA GAA CTG AGG CTG GAT GCT GAC OGiC ACA AAG AAA 786
Cys Ser Ile Leu Glu Glu Lau Arg Leu Asp Ala Asp AxgThr Lys Lys
135 140 145
GAT GAG CAA AAT CAA ATT CAA OOC ATG AGT CAC AGr GC.T TIC TTC AAC 834
Asp Glu Gln Asn Gln Ile G]n Pro Met Ser His Ser Ala Phe Phe Asn
150 155 160
AAG AAA ACA TTG AAC ACA GAA AOC AAT TGT GAA TAT AAiG GAC OCT GGG 882
Lys Lys Thr Leu Asn Thr Glu Ser Asn Cys Glu Tyr Lys Asp Pro Gly
165 170 175
AAA ATG ATT OOC AOG AGG OOC CAC CIT GGT TCT TCA CAG AAA CAA aCT 930
Lys blet Ile AxV Thr Arg Pro His Leu Ala Ser Ser GlnLys Gln Pro
180 185 190 195
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CAG AAA TGT TGC TTA TTP ACA GAA AGT TTG AAG C'I'G AAC GTA GAA GTG 978
Gln Lys Cys Cys Leu Phe Thr Glu Ser L8u Lys Leu Asn Leu Glu Val
200 205 210
AAC GGT CAG AAT GAA AGC AAT GAC ACA GAA CAG CIT GAT GAC (31T GTT 1026
Asn Gly Gln Asn Glu Ser Asn Asp Thr Glu Gln Leu Asp Asp Val Val
215 220 225
GGG TLT GGT CAG C,TA TTC AQC CAT AGC TCT TG`P GAT GOC TGC AQC AAG 1074
Gly Ser Gly Gln Lau Phe Ser His Ser Ser Ser Asp Ala Cys Ser Lys
230 235 240
AAT ATT CAT ACA OGA GAG ACA TZT TQC AAA GGT AAC CAG TM AGA AAA 1122
Asn Ile His Thr Gly Glu Ttr Phe Cys Lys Gly Asn Gln Cys Arg Lys
245 250 255
GTC ZGP GGC CAT AAA CAG TCA CTC. AAG CAA CAT CAA ATr CAT ALT CAG 1170
Val Cys Gly His Lys Gln Ser Leu Lys Gln His Gln Ile His Thr Gin
260 265 270 275
AAG AAA OCA GAT -GGA TgT TCr GAA T6'T GGG GOG AGC TTC ACC CAG AAG 1218
Lys Lys Pro Asp Gly Cys Ser Glu Cys Gly Gly Ser Phe Thr Gln Lys
280 285 290
Z'CA CAC CiC TTT GOC CAA CAG AGA ATT CAT AGT GTA GGA AAG CIC CAT .1266
Ser His Leu Phe Ala Gln Gln Arg Ile His Ser Val Gly Asn Leu His
295 300 305
GAA TGT GOC AAA TGT GGA AAA GOC TTC ATG OCA CAA C.'TA AAA C'TC AGT 1314
Glu Cys Gly Lys Cys Gly Lys Ala Phe Met Pro Gin Leu Lys Lsu Ser
310 315 320
GTATATCTGArA GATCATAGAGGTGATATAQOCZGTATATOCAAGGAA`a 1362
Val Tyr Lau Thr Asp His Thr Gly Asp Ile Pro Cys Ile Cys Lys Glu
325 330 335
4C3r GGG AAG GTC TTP ATT CAG AGA TCA GAA TI'G GRT ACG CAC CAG AAA 1410
Cys Gly Lys Val Phe Ile Gln Arg Ser Glu LSu Leu Thr His Gln Lys
340 345 350 355
ACA CAC AICT AIGA AAG AAG OOC TAT AAA TGC CAT GAC TGT GGA AAA GOC 1458
Thr His Thr Arg Lys Lys Pro Tyr Lys Cys His Asp GYs Gly Lys Ala
360 365 370
TTT TTC CAG AZG TTA TCT GTC TTC AGA CAT CAG AGA ALT CAC AGT AGA 1506
Phe Phe Gln Met Leu Ser Lau Phe Arg His Gln Arg Thr His Ser Arg
375 380 385
GAA AAA C'PC TAT GAA TM AGT GAA ZGT GGC AAA GGC TTC TGC CAA AAC 1554
Glu Lys Leu Tyr Glu Cys Ser Glu Cys Gly Lys Gly Phe Ser Gin Asn
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390 395 400
TCA ACC C'I'C ATT ATA CAT CAG AAA ATT CAT ACT GGT GAG AGA CAG TAT 1602
Ser Thr Leu Ile Ile His Gin Lys Ile His Thr Gly Glu Arg Gln Tyr
405 410 415
GCA TGC AAGr GAA ZCGT GOG AAA Gt7C TTT AOC CAG AAG TGA ACA G'TC AOC 1650
Ala Cys Ser Glu Cys Gly Lys Ala Phe Thr Gln Lys Ser Thr Leu Ser
420 425 430 435
TTG CAC CAG AGA ATC CAC TGA GGG CAG AAG TOC TAT GTG TGT MC GAA 1698
Leu His Gln Arg Ile His Ser Gly Gln Lys Ser Tyr Val Cys Ile Glu
440 445 450
TGC GGG CAG GOC TTC ATC CAG AAr GCA CAC CIG ATT GTC CAT CAA AGA 1746
Cys Gly Gin Ala Phe Ile.Gin Lys Ala His Uu Ile Val His Gln Arg
455 460 465
AGC CAC ACA GGA GAA AAA OCI' TAT CAG TOC CAC AAC TGT GGG AAA TOC .1794
Ser His Thr Gly Glu Lys Pro Tyr Gin Cys His Asn Cys Gly Lys Ser
470 475 480
TTC ATT TCC AAG TCA CAG CTT GAT ATA CAT CAT OGA ATT CAT ACA GGG 1842
Phe Ile Ser Lys Ser Gin Leu Asp Ile His His Arg Ile His Thr Gly -
485 490 495
GAG AAA OCT TAT GAA TOC AGT GAC TGT GGA AAA AOC TTC AOC CAA AAG 1890 -
Glu Lys Pro Tyr Glu Cys Ser Asp Cys Gly Lys Thr Phe Thr Gln Lys,
500 505 510 515
TCA CAIC CPG AAT ATA CAC CAG AAA ATT CAT ACr GGA GAA AGA CAC CAT 1938
Ser His Leu Asn Ile His Gin Lys Ile His Thr Gly Glu Arg His His
520 525 530
GTA TGC AGT GAA TOC GOG AAA GOC TTC AAC CAG AAG TCA ATA CTC AGC 1986
Val Cys Ser Giu Cys Gly Lys Ala Phe Asn Gln Lys Ser-Ile L8u Ser
535 540 545
ATG CAT CAG AGA ATT CAC ACC GGA GAG AAG C7CT TAC AAA TGC AGT GAA 2034
Met His Gln Arg Ile His Thr Gly Glu Lys Pro Tyr Lys Cys Ser Glu
550 555 560
T'GT GOG AAA GOC TTC AICT TOT AAIG TL`T CAA TZC AAA GAG CAT CAG C7GA 2082
Cys Gly Lys Ala Phe Thr Ser Lys Ser Gln Phe Lys Glu His Gin Arg
565 570 575
ATT CAC AOG GG'T GAG AAA CCC TAT GTG TaC ACT GAA TGT GOG AAG GOC 2130
Ile His Thr Giy Glu Lys Pro Tyr Val Cys Thr Glu Cys Gly Lys Ala
580 585 590 595
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TI'C AAC GGC AOG TCA MT TTC CAT AM CAT CAA ATA ACT CAC ACT AGA 2178
Phe Asn Gly Arg Ser Asn Phe His Lys His Gln Ile Thr His Thr Arg
600 605 610
GAG AGG CCT TTT GTC ZG'T TAC AAA TGT GGG AAG GCT TTT GTC CAG AAA 2226
Glu Arg Pro Phe Val Cys Tyr Lys Cys Gly Lys Ala Phe Val Gln Lys
615 620 625
TCA GAG TTG ATT ALJC CAT CAA AGA ejiCT CAC ATG GGA GAG AAA QOC TAT 2274
Ser Glu Lau Ile Thr His Gln Arg Thr His Met Gly Glu Lys Pro Tyr
630 635 640
GAA TM GTT GAC TGT GGG AAA TCG Tl'C AGr AAG AAA OCA CAA C'IC AAG 2322
Glu Cys Leu Asp Cys Gly Lys Ser Ptie Ser Lys Lys Pro Gln Leu Lys
645 650 655
(`a . CAT CAG CXaA ATT. CAC AOG GGA GAA AGA aCT TAT GTG TOT TCP GAA. 2370
Val His Gin Arg Ile His Thr Gly Glu Arg ProTyr Val Cys Ser Glu
660 665 670 675
ZGr GGA AAG GOC TTC AAC AAC AGG TCA AAC TTC AAT AAA CAC CAA ACA 2418
Cys Gly Lys Ala Phe Asn Asn Arg Ser Asn Phe Asn Lys His Gln Thr
680 685 690
ACT CAT AOC AGA GAC AAA TCT TAC AAA TGC AGT TAT TCT GTG AAA GOC 2466
Thr His Thr Arg Asp Lys Ser Tyr Lys Cys Ser Tyr Ser Val Lys Gly
695 700 705
TTT AOC AAG CAA TGAATICX,,'TA GrGCATCADC ATATTCATAA ATGAAATATA 2518
Phe Thr Lys Gln
710
CTC70CAGTTT CIRC'aAP,C'~,~AAG AGAACATGTP CPCAGAATCA GM"CPAATTA TA'tC~!'rATTG
2578
AATTCATGCT TCAGAAAAAC TCTAGGGATG CALTOCATgI' GTGAACAICAT GATAAAAAAG 2638
ZCATGCCI'TTA TTITAGTGAG GGCAATTACA GAGAAAAGAG TAAOCA~'aAAA TOI'LTIC'IG 2698
AGTAC'POGCC TCATTAAGGA TTATAAATTT TCI'OOOCGOG AAGAAA<JOCT GAiCTAACGC'A 2758
TTGAGAAAAG CGTPTCTG'rA AAC,AATGGTA CAAGACAOGT TGITAC'PaGA TTAT'ITATAG 2818
TAAAATATGT GGGAAATTAT ATCAATGATA ACOCIGTITA T'PGIGGC'ATA TCAATATTi'T 2878
TAAAOi'GCCA ACACAGI'CAT GATAGGGACAA TATTITATG'P GI'GPGR'G'IGC GCCiTA'PGTA 2938
TATAAGCATA TATATAATAT ATAAGCATAT TATTATATAC AGGTTGAGTA TOOCITCICC 2998
AAAATGuCTG GGATC:AGAAG CATTTTGGAT. TPCA(',ATACT TACAC'aATTIT GGAATATT'IG 3058
J*
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CATTATATTP ATTGG1'TC'iAG CATOOC,TAAT CIGAAAATOC AAGATTAP-AT GCPOCAATTA 3118
Gf.;ATTTOC'IT TC`AGOGICAT GTTAGAGTPC AAAAAGTT'I'C AGATTIRCW TTTPGAIGATT 3178
AGGAATAO(7C AAOCiGTATG TA+OGPATATT TCPQPATCPA TGTATGTATA TATATGOCATA 3238
TOC14GACATA TOPATATGGT CiGGPCAOCA TATGTGPATG TATGOGPATG TATOTATGTA 3298
TGTATGOOC.T CAGTGCAGT','Y' GGGI'I'TC~G'P(' CAL"'aAAT'PCAC TGCATAQCAG
GAC,ATGTAAG 3358
CAGATGAGIT ATPPPPTAAG AGAATLTAAT G'TAATIGTIT TTATAAAAAT TATTOOCPAT 3418
TGAATATRTA TATAA'1GAGG TWTATC,AAC AATGATTAAC 'POCTITATTA TA+CATAC,AC,A 3478
TGAAT I= TPTIC~PAA ATCaf'.ATAAAT GAi(',ATTCTAT 'AATOt'ITALT GATCPI'PATA 3538
TTAOAGATTT . TACy1f;,ATTAOC TCAOGTZ~pOC QOOOC7TTOC ATChC7C'AOCA 3598
TL'TATAGTGA G(7CTC1C7C'AT AATTAOI'C~C7C AAO(ATTAGT C'POGTI~CATA T'ITPTACAOC
3658
AOGAGICAAC AAACTGiGOC ATi'OOOCAAA TATOOOC,'TOC CAAGTGPI'TP TTTAAAATAA 3718
AGITTTATTG GAACACAAAA AAAAAAAAAA AAAAAA 3754
(2) I1vF0EN1ATI0N FOR SDQ ID N0:13:
(1) SEQ[1ENCE CTiARPiCPERISTICS:
(A) IENGTH: 389 aminu acids
(B) TYPE: amino acid
( D ) TOPO.LOW: linear
( ii ) NDLEC7[1LE TYPE: probe,in
( x3. ) SBQtJIINCE DES(32IPTIaNi: SF]Q ID Ia0:13 :
Met Ala Asp Pro Arg Asp Lys Ala Lau Gin Asp Tyr Arg Lys Lys Leu
1 5 10 15
Leu Glu His Lys Glu Ile Asp Gly Arg Leu Lys Glu Leu Arg Glu Gin
20 25 30
Leu Lys Glu Leu Thr Lys Gin Tyr Glu Lys Ser Glu Asn Asp Leu Lys
35 40 45
Ala Leu G1n.Ser Val Gly Gln Ile Val Gly Glu Val Leu Lys Gln LQu
50 55 60
Thr Glu Glu Lys Phe Ile Val Lys Ala Thr Asn Gly Pro Arg Tyr Vaal
CA 02458872 2004-03-23
65 70 75 80
Val Gly Cys RY+g Arg Gln Leu Asp Lys Ser Lys Leu Lys Pro Gly Thr
85 90 95
Arg Val Ala Lau Asp Met Thr Thr Leu Thr Ile Met Arg Tyr Leu Pro
100 105 110
Arg Glu Val Asp Pro Leu Val Tyr Asn Met Ser His Glu Asp Pro Gly
115 120 125
Asn Val Ser Tyr Ser Glu Ile Gly Gly Leu Ser Giu G]n Ile Arg Glu
130 135 140
Leu Arg Giu Val Zle Glu Leu Pro Lele Thr Asn Pro Glu Leu Phe Gln
145 150 155 160
Arg Val Gly Ile Ile Pro Pro Lys Gly Cys Leu Leu Tyr Gly Pro Pro
165 170 175
Gly Thr Gly Lys Thr Leu Leu Ala Arg Ala Val Ala Ser G]n Leu Asp
180 185 190
Cys Asn Phe Leu Lys Val Val Ser Ser Ser Ile Val Asp Lys Tyr Ile
195 200 205
Gly Glu Ser Ala Arg Leu Ile Arg Glu Met Phe Asn Tyr Ala Arg Asp
210 215 220
His Gin Pro Cys Ile Ile Pbe Met Asp Glu Ile Asp Ala I.l.e Gly Gly
225 230 235 240
Arg Arg Plhe Ser Glu Gly Thr Ser Ala Asp Arg Glu Ile Gln Arg Thr
245 250 255
Leu Met Glu Leu Leu Asn Gin Met Asp Gly Phe Asp Thr Lau His Arg
260 265 270
Val Lys Met Thr Met Ala Thr Asn Arg Pro Asp Thr Leu Asp Pro Ala
275 280 285
Leu Leu Arg Pro Gly Arg Leu Asp Arg Lys Ile His Ile Asp Leu Pro
290 295 300
Asn Glu Gin Ala Arg Leu Asp Ile Leu Lys Ile His Ala Gly Pro Ile
305 310 315 320
Thr Lys His Gly Glu Ile Asp Tyr Glu Ala Ile Val Lys Leu Ser Asp
325 330 335
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Gly Phe Asn Gly Ala Asp Lau Arg Asn Val Cys Thr Glu Ala Gly Met
340 345 350
Phe Ala Ile Arg Ala Asp His Asp Phe Val Val Gin Glu Asp Phe Met
355 360 365
Lys Ala Val Arg Lys Val Ala Asp Ser Lys Lys L2u Glu Ser Lys Leu
370 375 380
Asp Tyr Lys Pro Val
385 =
(2) INFORMATION FOR SEQ ID NO:14:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1167 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:14:
ATGOOGGAOC CTAGAGATAA GGflGC.TrCAG CAICPAUOOC',A AGAADTTGCT TGdo-ACAC'AAG 60
GAGATOGAOG GOOGTC.TrAA GGAGZTAAOG GAACAATTAA AAGAAGT'rAC CAAGCAGTAT 120
C',AAAAGIC,Z'G AAAATGATCT GAAGOOOLTA CAuGAGT(~'TTG GOCAC',ATaGT CaGGTGAAGTG
180
CTrAF-ACAQr TAA~.'rGAAC'A AAAATTCATT QITAAAGC.'TA CQhAZG1C'~QC AAGATATG'IT 240
GTuwTPGrC GTCGACAOCT TGHCAAAAGr AAGG'1'GAAOC CAGGAACAAG AGTrGG'ZTiG 300
GATATGACPA CACTAAC.TAT CATGAGATAT TrGOCGAGAG AGGIGGATaC ACZGGPiTAT 360
AACATGlCrC ATGAOGACOC TGGC'~ATM TCTPATTCiG AGATTOGAOG GCrATCAGAA 420
CAGAZbCGGG AATTAAGAGA GGIGATAGAA TTACC7rCITA CAAAOOCAGA GTTAT'ITCAG 480
WMAOC',~AA TAATACC'rCC AAAAGaCTGT Tr(~ITATATG GACCACCAGG TAOGGC'aAAAA 540
AlCAIC'tC.'rTOG CACGAGOOGT TGC'rAf30C14G CTOGACTGCA ATiTCITAAA OGPrGrATCT 600
AGTrCPATTG TAGACAAGrA CATrG(I'GAA AGrOGTCGZT TGATC.AGAGA AATGPTPAAT 660
TATGCTAGAIG ATCATCAAOC ATGCATCATT TTTAZGCaATG AAATAGATGC TATTGG7G(3T 720
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OGPOf~GTTTT CTGAGWPAC TTCAOCIGAC AGAGAGATTC AGAGAACG'IT AATGGAGITA 780
CTGAP-ZC,AAA ZGGARCaGATT TGATACTCiG CATAGAGZTA AAATC'õAOCAT GOC,TACAAAC 840
AGACCAGATA CALTGGP-TOC ZGiCPlRCdCiG CG1OCA3GAA GATTAGATAG AAAAATACAT 900
ATTGATITOC CAAATGAACA AGCAAGATTA GACATACTGA AAAZC7CATGC AG4PCCCATT 960
ACAAAGC'AT'G GiC'aAAATAGA TTATCaAAOCA ATTGPGAAOC TTI'CGGATGG CTTTAAZbC'aA 1020
GCAGATCZC'aA GAAATG'1'Tl'G TALTC'AAOC'A GGTATGITOG CAATPOM'GC TGATCATGAT 1080
TPIGTAGrAC AGGAAGACiT CAZCAAAQC"A GTCAGAAAAG TGC~C'i'GATTC TAAGAAGCi'G 1140
GAGPC.'TAAAT TOGACTAICAA AQChG'hG 1167
(2) INFORMATION FOR SEQ ID N0:15:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1566 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-331G07
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 17..1183
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:
GAGACGOCTT CZCATC ATG GOG GAC CCT AGA GAT AAG GOG CTP CAG GAC 49
Met Ala Asp Pro Arg Asp Lys Ala Leu Gln Asp
1 5 10
TAC CGC AAG AAG TTG CTT GAA CAC AAG GAG ATC GAC GOC CGT CTT AAG 97
Tyr Arg Lys Lys Leu Leu Glu His Lys Glu Ile Asp Gly Arg Leu Lys
15 20 25
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GAG TTA AGG GM C.AA TTA AAA GAA G'IT AOC AP-G CAG TAT GAA AAG TCT 145
Glu Leu Arg Glu Gln Leu Lys Glu Leu Thr Lys Gln Tyr Glu Lys Ser
30 35 40
GAA AAT GAT CTG AAG GflC C.TA CAG AGr GTT GGG CAG ATC G'PG GGT GAA 193
Glu Asn Asp Leu Lys Ala Leu Gin Ser Val Gly Gln Ile Val Gly Glu
45 50 55
GTG CIT AAA CAG TTA ACT GAA GAA AAA TTC ATT GTP AAA GGT A[JC AAT 241
Val Leu Lys Gln Leu Thr Glu Glu Lys Phe Ile Val Lys Ala Thr Asn
60 65 70 75
pGA CCA AGA TAT G'IT GTG GG1' WP OGP OGA CAG C1T GAC AAA AGr AArG 289
Gly Pro Arg Tyr Val Val Gly Cys Arg Arg Gln L8u Asp Lys Ser Lys
80 85 90
QiG AAG af:A. GGA ACA AGA G1T GCI' TTG GAT ATG ACT ACA CTA AICT ATC 337
Leu Lys Pro Gly Thr Axg Val Ala Leu Asp Met Thr Thr L8u Zhr-Ile
95 100 105
ATG AGA TAT TTG QOG AGA GAG GTG GAT CCA CPG GiT -TAT AAC ATG TLLT 385
Met Arg Tyr Leu Pro Arg Glu Val Asp Pro Leu Val Tyr Asn Met Ser
110 115 120
CAT GAG GAC OCT GOG AAT GTT TGT TAT TCT GAG ATT GGA GOG CTA TCA 433
His Glu Asp Pro Gly Asn Val Ser Tyr Ser Glu Ile Gly Gly Leu Ser
125 130 135
GAA CAG ATC COG GAA TTA AGA GAG GTG ATA GP,A TTA aGT CiT ACA AAC 481
Glu Gin Ile Arg Giu Leu Arg Glu Val Ile Glu Leu Pro Leu Thr Asn
140 145 150 155
CCA GAG TTA T'iT CAG OCT GTA GGA ATA ATA OCT CCA AAA GOC TGT TTG 529
Pro Glu L2u Phe Gin Arg Val Gly Ile Ile Pro Pro Lys Gly Cys Leu
160 165 170
TTA TAT GGA CCA CCA GGT AOG GGA AAA ACA CPC TTG OCA CGA GOC GTP 577
Leu Tyr Gly Pro Pro Gly Thr Gly Lys Thr Leu Leu Ala Arg Ala Val
175 180 185
GCT AQC CAG CTG GAC TGC AAT T1C TTA AAG GPr GTA = AGT WT ATT 625
Ala Ser Gin Leu Asp Cys Asn Phe Lau Lys Val Val Ser Ser Ser Ile
190 195 200
GPA GAC AAG TAC ATT GGr GAA AOr GCT OGT TTG ATC AGA GAA ATG TTT 673
Val Asp Lys Tyr Ile Gly Glu Ser Ala Arg Leu Ile Arg Glu Met Phe
205 210 215
AAT TAT GG'T AGA GAT CAT CAA aCA TGC ATC ATT TIT ATG GAT GAA ATA 721
Asn Tyr Ala Arg Asp His Gln Pro Cys Ile Ile Phe Met Asp Glu Ile
CA 02458872 2004-03-23
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220 225 230 235
GAT GL'T ATT GGT GGP CGT OGG TTT TGT GAG GGT AC,T 'ICA GCT GAC AGA 769
Asp Ala Ile Gly Gly Arg Arg Phe Ser Glu Gly Thr Ser Ala Asp Arg
240 245 250
GAG ATT CAG AGA AOG TTA ATG GAG TTA CTG AAT CAA ATG GAT GGA TZT 817
Glu Ile Gln Arg Thr Lau Met Glu Leu Leu Asn.Gin Met Asp Gly Phe
255 260 265
GAT AIC'!' CTG CAT AGA GTT AAA ATG AOC ATG OCT ACA AAC AGA a(:A GAT 865
Asp.Thr Leu His Arg Val Lys Met Thr Met Ala Thr Asn Arg Pro Asp
270 275 280
ACA CTG GRT aC'1' GCT TTG C1G a(OT OCA GGA AGA TTA GAT AGA AAA ATA 913
Thr Leu Asp Pro Ala Leu Lau Arg Pro Gly Arg Leu Asp Arg Lys Ile
285 290 295
CAT ATT GAT TTG CCA AAT GAA CAA GCA AGA TTA GAC ATA C1G AAA ATO 961
His ile Asp Leu Pro Asn Glu Gin Ala Arg Leu Asp Ile Leu Lys Ile
300 305 310 315
CAT GCA GGP OOC ATT ACA AAG CAT Gt31 GAA ATA GAT TAT GAA GCA ATT 1009
His Ala Gly Pro Ile Thr Lys His Giy Glu Ile Asp Tyr Glu Ala Ile
320 325 330
GTG AAG C'I'T TaG GAT GOC TTT AAT GCA GCA GAT CIG AGA AAT GiT TGT 1057
Val Lys Lsu Ser Asp Giy Phe Asn Gly Ala Asp Lau Arg Asn Val Gys
335 340 345
AGT GAA GCA GGT ATG TTC GCA ATT OGT GC.T GAT CAT GAT TTT GrA CTA 1105
Thr Glu Ala Gly Met Phe Ala Ile Arg Ala Asp His Asp Phe Val Val
350 355 360
CAG GAA GAC TTC ATG AAA GCA GTC AGR AAA GIG GGT GAT Tt.'T AAG AAG 1153
Gln Glu Asp Phe Met Lys Ala Val Arg Lys Val Ala Asp Ser Lys Lys
365 370 375
CTG GAG TG`P AAA TTG GAC TAC AAA OCT GIG TAATTPAL'ZG TAAGATTITP 1203
Leu Glu Ser Lys Leu Asp Tyr Lys Pro Val
380 385
GAT(0GC'tGCA TGAtCAGATGT TOOC'iTATTG TAAAAATAAA GTTAAAGAAA ATAATGTATG 1263
TAT'IGOC'.AAT GATGICATTA AAAGPATATG AATAAAAATA TGGAGrAAICAT CATAAAAATT 1323
AGTAATI'.AA CTI'I'TAAGAT ACACAP-GAAA TTTGPATGIT TGTTAAAGPP GCATTTATTG 1383
CAGCAAGTTA CAAAGGOGAAA GTGTTGAAGC TPTPCATATT TGC'IbOGPGA GCATri'TG"TA 1443
CA 02458872 2004-03-23
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AAATAT'I'f',AA ADt%'IT'iC ~A GATAGIGGTA TAAGAAAOC'A ZTTC'PTATGA CITATTTPC:'T
1503
ATCATPIK7TT TTC7C',PCATO'T AAAAAGPIC'aA ATAAAATC'IG TTI'GAZTC,AG TTCl'oC,'TAAA
1563
AAA 1566
(2) INFnFiMATION FUEt SDQ ID NO:16 :
( i ) SDQUENC'E CHARACTERISTICS:
(A) IE3VMH: 223 2Qa3no acids
(B) TYPE: 8mi,no acid
( D ) TOPOLOGY: litye3r
( ii ) M3IEC.UIE TYPE: protBin
( xi ) SBOI)IIN.'E DFSCRIPTION: SBQ ID 1V0:16 :
MBt Ser Asp Glu Glu Ala Arg Gin Ser Gly Gly Ser Ser Gin Ala Gly
1 5 10 15
Val Val Thr Val Ser Asp Val Gln Glu L$u Met Arg Arg Lys Glu Glu
20 25 30
Ile Giu Ala Gln Ile Lys Ala Asn Tyr Asp Val Leu Glu Ser G1n.Lys
35 40 45
Gly Ile Gly Met Asn Glu Pro Leu Val Asp Cys Glu Gly Tyr Pro Arg
50 55 60
Ser Asp Val Asp Leu Tyr Gln Val Arg Thw Ala Arg His Asn Ile Ile
65 70 75 80
Cys Leu Gln Asn Asp His Lys Ala Val Met Lys Gln Val Glu Glu Ala
85 90 95
Leu His Gln Leu His Ala Arg Asp Lys Glu Lys Gln Ala Arg Asp Met
100 105 110
Ala Glu Ala His Lys Glu Ala Met Ser Arg Lys Leu Gly Gln Sear Glu
115 120 125
Ser Gln Gly Pro Pro Arg Ala Phe Ala Lys Val Asn Ser Ile Ser Pro
130 135 140
Gly Ser Pro Ala Ser Ile Ala Gly Leu Gin Val Asp Asp Glu Ile Val
145 150 155 160
Glu Phe Gly Ser Val Asn Thr Gln Asn Phe Gln Ser Leu His Asn Ile
CA 02458872 2004-03-23
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165 170 175
Gly Ser Val Val Gln His Ser Glu Gly Lys Pro Leu Asn Val Thr Val
180 185 190
Ile Arg Arg Gly Glu Lys His Gln Leu Arg Leu Val Pro Thr Arg Trp
195 200 205
Ala Gly Lys Gly Leu Lau Gly Cys Asn Ile Ile,Prc Leu Gln Arg
210 215 220
(2) INFORMATION FOR SEQ ID NO:17:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 669 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D). TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:
ATGTCCGACG AGGP-AG(X'~G GCA~C'aAl00GC'~ GGCTOC'POGC ~r CGTGAL`lGIC 60
AOC('~?lOGTM AGGAIOCTC'aAT GCGOOGCAAG GAGGAGATAG AAGOGCAGAT CAAGGOC',AAC 120
TATGACGPGC ZGC,hAAOOCA AAAAOGCATT GGGATGAACG A CCGCTGOT GGAICTCOlGAG 180
GGCTAO000C GGPCAGACGT GGACCiOPAC CAAG'i'OOOC',A CCGOCAOGCA CAACATCATA 240
TGOC'!'GCAiGA ATGATCACAA aOCAG'Z+C'~ATG APuGCAGGTGG AGGAi0GO0CP GCACCAOC.'TG
300
CAOGCTC'XGCG ACAAOC'~A (~('1LGGOOCGG GACATGGC,'TG AGOCOCACAA AGAGGYJC.ATG 360
A COGC',AAAC TGGGTCAGAG TGAC'~AGCCAG GGOCCPOCAC GGGCCrPCGC CAAAG'iC'aAAC 420
AQCAZ'CAOOC OL~G C'i'OOOC AGbCAGCATC GOGl3C~'TCPGC AAG'rOGATGA TGAGRTTGiG 480
GAC~ITOOGCP ClGIC'~AACAC CC'.ACAACTPC CAGTCACTOC ATAACATTGG CAtGPGrGGTG 540
CAGCACAGIG AGGCGGC'~CC CCTGAATCOPG ACAGI'GATCC GCAOGGGOGA AAAACACCAG 600
CTTAGACTPG Ti'OCAACACG CPGGGCAGGA AAAGGAL'PGC TGGGCI'GCAA CATTATTOCP 660
CTOCAAAGA 669
CA 02458872 2004-03-23
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(2) INFORMATION FOR SEQ ID NO:18:
(1) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1128 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: CDNA
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
.(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-163D09
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 125..793
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:
ACIC~imCi+OG OOI*I'0 OG('~A OOOCPGl'OGT GTTTTGGOOC ATGGGC00C'aAG OGTAGiTACG 60
G71'CC'~CTOOG GQIC)GIOOC TAGOOOOOGA GCOGGGTL`rC TGGAG7.`OGOG GOOOOOOGIT 120
CAOG ATG TOC GAC GAG GAA GOG ApG CAG ROC GGA GOC TUC RC)G CAG GOC 169
Met Ser Asp Glu Glu Ala Arg G1n Ser Gly Gly Ser Ser Gin Ala
1 5 10 15
GOC OW GTG ALT GTC AOC GAC G'M CAG GAG CTG ATG COG COC A14G GAG 217
Gly Val Val Thr Val Ser Asp Val Gln Glu Lau Met Arg Arg Lys Glu
20 25 30
GAG ATA GAA GC7G CAG ATC AAG GOC AAC TAT GAC GTG CTG GAA AQC CAA 265
Glu Ile Glu Ala Gln Ile Lys Ala Asn Tyr Asp Val Leu Glu Ser Gln
35 40 45
AAA GGC ATT GGG ATG AAC GAG COG CTG GPG GAC TGT GAG GOC TAC QOC 313
Lys Gly Ile Gly Met Asn Glu Pro Leu Val Asp Cys Glu Gly Tyr Pro
50 55 60
COG TCA GAC GTG GAC CTG TAC CAA GTC aGC ACC GL7C AGO CAC AAC ATC 361
Arg Ser Asp Val Asp Leu Tyr Gln Val Arg Thr Ala Arg His Asn Ile
65 70 75
ATA TGC CTG CAG AAT GAT CAC AAG GCA GTG ATG AAG CAG GTG GAG GAG 409
CA 02458872 2004-03-23
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Ile Cys Leu Gln Asn Asp His Lys Ala Val Met Lys Gln Val Glu Glu
80 85 90 95
GOC CIG CAC CAG CTG CAC GCT t)GC GAC AAG GAG AAG CAG GOC CGG GAC 457
Ala Lsu His Gln Lau His Ala Arg Asp Lys Glu Lys Gln Ala Arg Asp
100 105 110
ATG GCT GAG GOC CAC AAA GAG G{7C ATG AGC OGC AAA CIG GGT CAG AGT 505
Met Ala Glu Ala His Lys Glu Ala Met Ser Arg Lys Leu Gly Gln Ser
115 120 125
GAG AGC CAG GGC OCT CCA CGG GOC TTC GOC AAA GTG AAC AGC ATC AGC 553
Glu Ser Gln Gly Pro Pro Arg Ala Phe Ala Lys Val Asn Ser Ile Ser
130 135 140
0 C GGC TOC af.'A GOC AGC ATC GOG GOT CTG CAA GIG GAT GAT GAG ATT 601
Pro Gly Ser Pro Ala'Ser Ile Ala Gly Leu Gln Val Asp Asp Glu Ile
145 150 155
C31G GAG TTC GOC TCT GTG AAC ACC CAG AAIC TTC CAG TCA CIG CAT AAG 649
Val Glu Phe Gly Ser Val Asn Thr Gln Asn Phe Gln Ser Leu His Asn
160 165 170 175
ATT GGC AGP GTG GIG CAG CAC AOT GAG GGG AF-G OOC CIG AAT GTG ACA 697
Ile Gly Ser Val Val Gln His Ser Glu Gly Lys Pro Leu Asn Val Thr
180 185 190
GTG ATC aGC AGG GGG GGAP, AAA CAC CAG CIT AGA C'iT GTT aCA ACA OSC 745
Val Ile Arg Arg Gly Glu Lys His Gln Leu Arg Leu Val Pro Thr Arg
195 200 205
TGG GCA GGA AAA GGA CIG CTG GOC TOC A14C ATT ATT OCT CTG CAA AGA 793
Trp Ala Gly Lys Gly Leu Lsu Gly Cys Asn Ile Ile Pro Leu Gin Arg
210 215 220
TGATTC#iK70C TGGO['AAOAG TAACl410GAAA GCATCTTOOC TTGOOCIC'GCGA C'1'i'GwlC`PA
853
GGGATIT,OCA ACTTQIC."TTC TCPOOCIGAA GCATAAGGAT CTGGAAGAGG CT'iGTAAOGT 913
GAACTTCTGT GTOGPGGCAG TAC.'TGPOGOC CAOCAG'PCOPA ATC'i'OOCI'GG ATTAAGOCAT 973
TGRTAAAAAC TTAGGCPt'GG C7C7PC'ITiCAC AAATTAGGC7C AOGGOOC.'TAA ATAGC',AATTC
1033
aCTOC'~-TTG GGGCAAG'I'GG GOaGAAOPTA TPCIGGCAOG TAAC'PGGTGTG ATTATTATTA 1093
TTA'PTTTPAA TAAAGAGTTT TACAGTGC'I'G ATATG 1128
(2) INFQFiMATION FOR SEQ ID N0:19 :
CA 02458872 2004-03-23
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( i ) SEQUENCE CHARACTERMICS:
(A) LFNG'i'H : 506 acRi no acids
(8) TYPE: amizb acid
( D ) TOPOLOGR: linear
( ii ) MOL CIILE TYPE: protein
( xi ) SEQi7IINCE DESCRIPTICN: SH]Q ID N0:19 :
Met Ala Glu Ala Asp Phe Lys Met Val Ser Glu Pro Val Ala His Gly
1 5 10 15
Val Ala Glu Glu Glu Met Ala Ser Ser Thr Ser Asp Ser Gly Glu. Glu
20 25 30
Ser Asp Ser Ser Ser Ser Ser Ser Ser Thr Ser Asp Ser Ser Ser Ser
35 40 45
Ser Ser Thr Ser Gly Ser Ser Ser Gly Ser Gly Ser Ser Ser Ser Ser
50 55 60
Ser Gly Ser Thr.Ser Ser Arg Ser Arg Leu Tyr Arg Lys Lys Arg Val
65 70 75 80
Pro Glu Pro Ser Arg Arg Ala Arg Arg Ala Pro Leu Gly Thr Asn Phe
85 90 95
Val Asp Arg LSU Pro Gln Ala Val Arg Asn Arg Val Gln Ala Lau Arg
100 105 110
Asn Ile Gln Asp Glu Cys Asp Lys Val Asp Thr Leu Phe Leu Lys Ala.
115 120 125
Ile His Asp Leu Glu Arg Lys Tyr Ala Glu Leu Asn Lys Pro Lau Tyr
130 135 140
Asp Arg Arg Phe Gin Ile Ile Asn Ala Glu Tyr Glu Pro Thr Glu Glu
145 150 155 160
Glu Cys Glu Trp Asn Ser Glu Asp Glu Glu Phe Ser Ser Asp Glu Glu
165 170 175
Val Gln Asp Asn Thr Pro Ser Glu Met Pro Pro Leu Glu Gly Glu Glu
180 185 190
Glu Glu Asn Pro Lys Glu Asn Pro Glu Val Lys Ala Glu Glu Lys Glu
195 200 205
Val Pro Lys Glu Ile Pro Glu Val Lys Asp Glu Glu Lys Glu Val Ala
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210 215 220
Lys Glu Ile Pro Glu Val Lys Ala Glu Glu Lys Ala Asp Ser Lys Asp
225 230 235 240
Cys Met Glu Ala Thr Pro Glu Val Lys Glu Asp Pro Lys Glu Val Pro
245 250 255
Gln Val Lys Ala Asp Asp Lys Glu Gln Pro Lys Ala Thr Glu Ala Lys
260 265 270
Ala Arg Ala Ala Val Arg Glu Zhr His Lys Arg Val Pro Glu Glu.P,zg
275 280 285
LQu Arg Asp Ser Val Asp Letz Lys Arg Ala Arg Lys Gly Lys Pro Lys
290 295 300
Arg Glu Asp Pro Lys Gly Ile Pro Asp Tyr Trp Leu Ile Val Lau Lys
305 310 315 320
Asn Val Asp Lys Lsu Gly Pro Met Ile Gln Lys Tyr Asp Glu Pro Ile
325 330 335
Leu Lys Phe Leu Ser Asp Val Ser Leu Lys Phe Ser Lys Pro Gly Gln
340 345 350
Pro Val Ser Tyr Thr Phe Glu Phe His Phe Leu Pro Asn Pro Tyr Phe
355 360 365
Arg Asn Glu Val Leu Val Lys Thr Tyr Ile Ile Lys Ala Lys Pro Asp
370 375 380
His Asn Asp PFro Phe Phe Ser Trp Gly Trp Glu Ile Glu Asp Cys Lys
385 390 395 400
Giy Cys Lys Ile Asp Arg Arg Arg Gly Lys Asp Val Thr Val Thr Thr
405 410 415
Thr Gln Ser Arg Thr Thr Ala Thr Gly Glu Ile Glu Ile Gin Pro Arg
420 425 430
Val Val Pro Asn Ala Ser Phe Phe Asn Phe Phe Ser Pro Pro Glu Ile
435 440 445
Pro Met Ile Gly Lys L8u Giu Pro Arg Glu Asp Ala Ile Leu Asp Glu
450 455 460
Asp Phe Glu Ile Gly Gln Ile Leu His Asp Asn Val Ile Leu Lys Ser
465 470 475 480
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Ile Tyr Tyr Tyr Thr-Gly Glu Val Asn Gly Thr Tyr Tyr Gln Phe Gly
485 490 495
Lys His Tyr Gly Asn Lys Lys Tyr Arg Lys
500 505
(2) INFORMATION FOR SEQ ID NO:20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1518 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:
AT(30CAGIAAG CAGATITPAA AA7GU1'CTQG GAACCIGIC7G CX7C'A7npp(;P ZGOCGAAGAG 60
GAGARCiOCPA GCTOGALTAG ZGATd'C.ZGGG. GAAGAATCTG AIC'AGCAGTAG CTCTAGC'AGC 120
PGCACrAG3G ACAGC'1DC'.AC'' CAiOC',AIOCAOC ACTAGi'GOC'A GCAlOCAGCOG CAOCGGCAGC
180
AGCAOC'.AOCA GCAGCGGCAG CACTAOC'AOC CGC,AG000CT TOrATAC',A GAAG140GCTA 240
OCTGA(~OC'IT CCAGAAOOGC GODU0MOC CCGPiGOC,AA CAAATT'IC7GT GGATAGOCIG 300
CfCTCAGGCAG TTAGAAATCG TGZGCAAOOG CTPAGAAA~(`.A 4TCAA~('õATGA ATGt'GGAC,AAG
360
OPAGATACOC TGl'PCITAAA AGCAATI'CAT GAT(,'TTf'mAAA GkAAATATGC TGAACIrAAC 420
AAGCCt`CI'GV ATGA.TAGG0(" GT'IZ'CAAATC ATC'.AATGC',AG AATACGAGOC TACAC'aAAGAA
480
GAATGTt`=AAT GGAATP('1~1G& GGATGAGC"aAG TTCFuOCAGIG ATGAGGAiGGT GCAOC'aATAAC
540
A+GOC7CPAGZG AAATGCCt"OC CTTAGAOGQr GAOC'~AA('~AG AAAA00(.~TAA AGAAAAOCC'A 600
GAGGTCkAAG CTGAAGAiC,AA GGAAGI"IC)CT AAAC'aAAATTC CTGAGGl'('aAA GGATGAAGAA 660
AAiOGAAGTIG CTAAAC'1kAAT TC7CTC'rAiOGTA AAGGCTGA+-G AAAAAGCACaP- TTCPAAAGAC
720
TG'TATGGAOG CAAOCCCI'GA AGTAAAAGAA GATCCPAAAG AAG''TOCCCCA GGTAAApG('.A 780
GATGATAAAG AACAGOGTAA AOCAP-CAGAG GCTAAQOCAA GGOC.`PGCAGT AAIGA~"~At"'ACT 840
CATAAAAGAG TTOCPGAGC'~A AAGGCTIYJGG GACAG'TGTAG A'iCTTAAAAG A+OCI'AOGAAG 900
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-130-
GGAAADCCTA AAAGAGAAGA COCTAAAGGC ATTOCrGACP AZTGOCTGAT TO`PITPAAAG 960
AAZCTTGACA AGCTOOOGOC TATG:ATir'.,AG AAGTATGATG AOOOCATTCT GAAiG'T`t'CPiG 1020
TOGGATGTTA GOCTGAAG'iT CrCAAAACL."T GGOf`~GOLTG TAF-GITACAC CZTPGAATTT 1080
CATTTTGTAC OCAAOOCATA CPTCA~''aAAAT GAGvR~GCR~C~G TGAAGAIC'ATA TATAATAAAG 1140
GCAAAAC'AG A'L^CAMTGA '!'OCC.'TTCPTI' ZCTIC~OOC'aAT GGGC'aAFiAT'i'GA
AGATTGCAAA 1200
CGCIGCAAGA TAGAOOGGAG AACaAGGAAAA GATO1TAC1G TGACAACtAC CCAGAGT00C 1260
-ACAACIGGTA CIGGWAAAT.'iGAAATOC,A[' (7CAA["aAGROG TTOCPAATGC ATCATTCIZC 1320
AAC1't'C7[TTA GI~OG9!OCi~GA GAZ'i'OGrATG ATPOOGAAOC TGGAAOC.AOC' AGAAGF,TGCT
1380
ATCCPGC'aA'!G AOGALR'TTGA AATPGGOCAG ATTITACA'I"G ATAATQiC'AT 0CIGAATCA 1440
ATCTATTACT ATALTGGAGA AG'iC'AATGGT AC(TACTATC AA7TI'OGCAA ACATTATGGA 1500
AAiCAAGAAAT ACAGAAAA 1518
(2) INFORMATION FOR SEQ ID NO:21:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2636 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(lv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-078D05
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 266..1783
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:21:
GATTOGOC.`IG CGGPAICATCT aGGCACPCTA GC'POCAGOOG GGAGAGGOCT TGUOGCCAaC 60
aC'1'GPC7G'OOC AAGOCTOC'AC TGCOGCTGCC ACC't'CAOOOC CGG0CTC'PGC ATC7COCAGCT 120
CA 02458872 2004-03-23
-131-
OCAGC'1'OOOC ZC;tO00000C TGCZGCC7CATC GOOGCT'GCCA aCPCX7GC'.AQC (X70GOCCI'aC
180
GOOOCOOCCA CCCAApCATC CGTGAGICAT TTTCI'GOOCA TCPCIC~G'POG OOOOGPC'I'OC 240
CZGGPAIGAGP TTafAGGCiT GGAAG ATG GCA GAA GCA GAT TTP AAA ATG GTG 292
Met Ala Glu Ala Asp Phe Lys Met Val
1 5
TOG GAA CCP GTC GUC CAT GGG G1T GCC GAA GAG GAG ATG GCT AGC TCG 340
Ser Glu Pro Val Ala His Gly Val Ala Glu Glu Glu Met Ala Ser Ser
15 20 25
ACT AGT GAT TLT GOG GAA GAA TLT GAC AGC AGT AGC TCT AGC AGC AGC 388
Thr Ser Asp Ser Gly Glu Glu Ser Asp Ser Ser Ser Ser Ser Ser Ser
30 35 40
ACT AGT GdAC AOC AOC AGC AGC AGC AOC ACT AGT GGC AGC AGC AG1C GOC 436
Thr Ser Asp Ser Ser Ser Ser Ser Ser Thr Ser Gly Ser Ser Ser Gly
45 50 55
AQC GGC AGC AOC AOC AOC AGC AGC OOC AGC ACT AGC AOC OOC AOC CGC 484
Sex' Gly Ser Ser Ser Ser Ser Ser Gly Ser Thr Ser Ser Arg Ser Arg
60 65 70
TIPG TAT AGA AAG AAG AGG GrA OCT GAG OCT TCC AGA AGG GCG COG COG 532
L8u Tyr Arg Lys Lys Arg Val Pro Glu Pro Ser AYg Arg Ala Arg Arg
75 80 85
Gt7C OOG TTG GGA ACA AAT TTC GTG GAT AGG CPG C7CT CAG GCA OIT AGA 580
Ala Pro Leu Gly Thr Asn Phe Val Asp Arg Leu Pro Gin Ala Val Arg
90 95 100 105
AAT Q(T Gi'G CAA GCG CIT AGA AAC ATT CAA GAT GAA TGT GAC AAG GTA 628
Asn Arg Val Gin Ala Leu Arg Asn Ile Gln Asp Glu Cys Asp Lys Val
110 115 120
GAT AiOC CIG TTC TTA AAA GCA ATT CAT GAT CIT GAA AGA AAA TAT GCT 676
Asp Thr Leu Phe Lau Lys Ala Ile His Asp L8u Glu Arg Lys Tyr Ala
125 130 135
GAA CTC AAC AAG CCT CTG TAT GAT AGG O G TTT CAA ATC ATC AAT GCA 724
Giu Leu Asn Lys Pro Leu Tyr Asp Arg Arg Phe Gln Ile Ile Asn Ala
140 145 150
GAA TAC GAG OC'1' ACA GAA GAA GAA TGI' GAA TGG AAT TCA GAG GAT GAG 772
Glu Tyr Glu Pro Thr Glu Glu Glu Cys Glu Trp Asn Ser Glu Asp Glu
155 160 165
GAG TTC AGC AGT GAT GAG GAG GTG CAG GAT AAC ACC CCT AGT GAA ATG 820
Glu Phe Ser Ser Asp Glu Glu Val Gln Asp Asn Thr Pro Ser Glu Met
CA 02458872 2004-03-23
-132-
170 175 180 185
CCT OOC TTA GAG GGT GAG GAA GAA GAA AAC CCT AAA GAA AAC aCA GAG 868
Pro Pro Lp-u Glu Gly Glu Glu Glu Glu Asn Pro Lys Glu Asn Pro Glu
190 195 200
GTG AAA GCT GAA GAG AAG GAA GTP aCT AAA GAA ATT OCT GAG GTG AAG 916
Val Lys Ala Glu Glu Lys Glu Val Pro Lys Glu Ile Pro Glu Val Lys
205 210 215
GAT GAA GAA AAG GAA GTT GCT AAA GAA ATT aCT GAG GTA AAG GCT GAA 964
Asp Glu Glu Lys Glu Val Ala Lys Glu Ile Pro Glu Val Lys Ala Glu
220 225 230
GAA AAA OCA GAT TZ:T AAA GAC TGT ATG GAG GCA ACC OCT GAA GTA AAA 1012
Glu Lys Ala Asp Ser Lys Asp Cys Met Glu Ala Thr Pro Glu Val Lys
235 240 245
GAA GAT OCI' AAA GAA GTC OOC CAG GTA AAG GC'A GAT GAT AAA GAA CAG 1060
Glu Asp Pro Lys Glu Val Pro Gln Val Lys Ala Asp Asp Lys Glu Gln
250 255 260 265
CCT AAA GCA ACA GAG GCT AAG GCA AGG GLT GCA GTA AGA GAG AICT CAT 1108
Pro Lys Ala Thr Glu Ala Lys Ala Arg Ala Ala Val Arg Glu Thr His
270 275 280
AAA AGA GiT CCT GAG GAA AGG GTP OGG GAC AGT GTA GAT CiT AAA AGA 1156
Lys Arg Val Pro Glu Glu Arg Leu Arg Asp Ser Val Asp Lau Lys Arg
285 290 295
GCT AIOG AAG GGA AAG aCT AAA AGA GAA GAC OCT AAA GGC ATT OCT GAC 1204
Ala Arg Lys Gly Lys Pro Lys Arg Glu Asp Pro Lys Gly Ile Pro Asp
300 305 310
TAT TOG C1G ATT GTT TTA AAG AAT GTT GAC AAG CIC GOG OCT ATG ATT 1252
Tyr Trp Leu Ile Val Leu Lys Asn Val Asp Lys Leu Gly Pro Met Ile
315 320 325
CAG AAIG TAT GAT GAG OOC ATT G"IG AAG TTC TTG TOG GAT GTT AGC CIG 1300
Gin Lys Tyr Asp Glu Pro Ile Leu Lys Phe Leu Ser Asp Val Ser Leu
330 335 340 345
AAG TTC TCA AAA OCT GGC CAG OCT GTA AGT TAC AOC TTP GAA TTT CAT 1348
Lys Phe Ser Lys Pro Gly Gln Pro Val Ser Tyr Thr Phe Glu Phe His
350 355 = 360
TTT CTA COC AAIC OCA TAC TTC AGA AAT GAG GTG CTG GTG AP-G ACA TAT 1396
Phe Leu Pro Asn Pro Tyr Phe Arg Asn Glu Val Leu Val Lys Thr Tyr
365 370 375
CA 02458872 2004-03-23
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ATA ATA AP-G GCA AAA C7CA GAT CAC MT GAT QOC TTG TTT Tt.T TGG GGA 1444
Ile Ile Lys Ala Lys Pro Asp His Asn Asp Pro Phe Phe Ser Trp Gly
380 385 390
TGG GAA ATT GAA GAT TGC AAA GOC TGC AAG ATA GAC COG AGA AGA GGA 1492
Trp Glu Ile Glu Asp Cys Lys Gly Cys Lys Ile Asp Arg Arg Arg Gly
395 400 405
AAA GAT GTT A+CT GTG ACA ACT AOC CAG AGT GGL ACA ACT GCT ACT GGA 1540
Lys Asp Vai Thr Val Thr Zhr Thr Gln Ser Arg Thr Thr Ala Thr Gly
410 415 420 425
GAA ATT GAA ATG CAG CxA AGA GIG GT'T OCT AAT GCA TCA TTC TTC AAC 1588
Glu Ile Glu Ile G1n-Pro Arg Val Val Pro Asn Ala Ser Phe Phe Asn
430 435 440
TTC TTT AGT OCT GCT.GAG ATT C7CT ATG ATT GOG AAG CTG GAA aCA CJGA 1636-
Phe Phe Ser Pro Pro Glu Ile Pro Met Ile Gly Lys Leu Glu Pro Arg
445 450 455
GAA GAT GCT ATC CTG GAT GAG GAC TTT GAA ATT GGG CAG ATT TTA CAT 1684
Giu Asp Ala Ile Leu Asp Glu Asp Phe Glu Ile Gly Gln Ile Leu His
460 465 470
GAT AAT GTC ATG C'IG AAA TCA ATC TAT TAC TAT ACT GGA GAA GTC AAT 1732
Asp Asn Val Ile Leu Lys Ser Ile Tyr Tyr Tyr Thr Gly Glu Val Asn
475 480 485
GGT ACC TAC TAT CAA TZT GOC AAA CAT TAT GGA AAC AAG AAA TAC AGA 1780
Gly Thr Tyr Tyr Gln Rwe Gly Lys His Tyr Gly Asn Lys Lys Tyr Arg
490 495 500 505
AAA. TAAiC~'PC.AATC TGAAAGATZT TTCAAGIATC TTAAAATCf C AA~'aAAGTC'aAA 1833
Lys
GCAGATTCAT ACAOOC.`I'!GA AAAAAGPAAA AOOC'I'GAOLT GTAAGCTGAA CACITATTATT 1893
aGTTATAGIC AAG'1'TTT'1GT GGPTTCTPuG TAlGTCTATAT TTTAAAAATA GTCC.TAAAAA 1953
GTGTCTAAGT GOCRGTi'TAT TCTATGTAGG ('TGPT(`TAGT ATAATATTG'1' TC'.~1AAATATG 2013
TAAGC7."GTIG TCAATTATGT AAAGC'ATG<I'T AGTTTGGTTW TAICAC'AGTGP TGAZTTPPGT 2073
GATGTOCPTT GG'iCATGI"iT CTGTTAGACT GTAGCTG'1'GA AACTG7CAGA AT'i'GTTAAL'T 2133
GAAACAAATA TTTGCTIC'aAA AAAAAAAGTT CATGAAGTAC CAATCI,AAGT GTT'iTATTTT 2193
T'IT1C'ITPtT 'I'OC'.AGOOCAT AAGAC,TAAOG GTITAAATCr GCITGCA~.'TA GCI'GIGOClV
2253
CAZTAGTTTG CTATAGAAAT OCAGTAGTTA TAIGTAAATAA AACAGiGTAT TTWAAGTI'P 2313
CA 02458872 2004-03-23
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GACI'GCTZGA AAAAOhTTAG CATACA'I'C.TA A'ICRC'aAAAAG AIQCACATTPG ATTC;AAGTGA
2373
GAaC'PI'QPGP ATGZY'ACATA TAGTOGOCTA TAAATTTAAT CATAATGATG TTAT'iGT7TA 2433
OCAICTGAGGT GTTAATATAA CATAGTkPIT TPGAAAFsAGP T7CT'ICAT{,`P TATATTGIGP 2493
AATTGTAAPC TAAAGATA+OC G'IvTPTTC7IT TGTATL`QTG'P TGTAOCTPOC CITTCACrGA 2553
AAATGATCAC TT~AT'ITGAT RCi'GITPITC ATG'PI'CTl'GP ATZGCAAQCT AAAATAAATA 2613
AATATTAAAG TGTGITATAC TAT 2636
(2) INFIOWATION FOR SEQ ID N0: 22 :
( i ) sDQ[1BNCE CWUUCTERLSTICS:
(A) LENC#iH: 170 amino acids
( B ) TYPE: aatirm acid
( D ) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SnQ[IF.~VCE DESCRIPTION: SDQ ID N0: 22:
Met Thr Glu Leu Gln Ser Ala Leu Leu Leu Arg Arg Gin Leu Ala Glu
1 5 10 15
Leu Asn Lys Asn Pro Val Glu Gly Phe Ser Ala Gly Leu Ile Asp Asp
20 25 30
Asn Asp Leu Tyr Arg Trp Glu Val Leu Ile Ile Gly Pro Pro Asp Thr
35 40 45
Lsu Tyr Glu Gly Gly Val Phe Lys Ala His L2u Thr Phe Pro Lys Asp
50 55 60
Tyr Pro Leu Arg Pro Pro Lys Met Lys Phe Ile Thr Glu Ile Trp His
65 70 75 80
Pro Asn Val Asp Lys Asn Gly Asp Val Cys I1e Ser Ile Leu His Glu
85 90 95
Pro Gly Glu Asp Lys Tyr Gly Tyr Glu Lys Pro Glu Glu Arg Trp Leu
100 105 110
Pro Ile His Thr Val Glu Thr Ile Met Ile Ser Val Ile Ser Met Leu
115 120 125
Ala Asp Pro Asn Gly Asp Ser Pro Ala Asn Val Asp Ala Ala Lys Glu
CA 02458872 2004-03-23
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130 135 140
Trp Arg Glu Asp Arg Asn Gly Glu Phe Lys Arg Lys Val Ala Arg Cys
145 150 155 160
Val Arg Lys Ser Gin Glu Thr Ala Phe Glu
165 170
(2) INFORMATION FOR SEQ ID N0:23:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 510 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:
ATC'aACGGAOC TGCAGZ'00GC AC'POCTALTG CGAAGAC',AOC RCGCALdAPdLT CAACAAAAAT 60
CCAG7I'GGGAAG GCTiTI'G`IC~C AGGTZTAATA GATGACAATG ATCTCPACCG ATOGGAAGIC 120
CTTATTATTG GCOC'i'CCAGA TAC.ACTZTAT GAAGG1'GGTG TTl'TTAA<3GC TCATLRTACT 180
T'iC)CC.AAAAG ATTATCCOCP CCGACCTCCT AAAATGAAAT TCATTACAC'xA AATCTGGCAC 240
CCAAATGTTC ATAAAAATGG TGAT I'GTGC ATl'TCTATTC TTCAZOAGCC ZGGGCoAAGAT 300
AAGTATGUT'P ATGAAAAOOC AG'AGGAAOOC TOOC.TOOCPA TCG9CACTGP GGAAAOCATC 360
ATGATTAGIG TCAT=AT GCTGGCAGAC CCTAAZbC'~(' ACl'CAUCTGC TAATGT'1'GAT 420
GCPOOC'~AW AATGGAGGGA AGATAGAAAT GGAGAATTTA AAAGAAAAOr TGCOOO= 480
GTAAGAAAAA GC7GAAGAiGAiC TGCPTTIC'aAG 510
(2) INFORMATION FOR SEQ ID NO:24:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 617 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
CA 02458872 2004-03-23
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(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-423A12
(is) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 19..528
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:
GGGOCCfOG(' CAGGC'AM ATG AC7G GAG CTG CAG 'hOG GCA CI+G CTA CiG CGA .51
Met Thr Glu Leu Gln Ser Ala Leu Lau Leu Arg
1 5 10
AGA CAG CTG GCA GAA C3'C AAC AAA AAT OCA GTG GAA GOC TTT TC'T GCA 99
Arg Gin Lau Ala Glu Leu Asn Lys Asn Pro Val Glu Gly Phe Ser Ala
15 20 25
GGT TTA ATA GAT GAC AAT GAT CTC TAC OGA TGG GAA GTC CTT ATT ATT 147
Gly Leu Ile Asp Asp Asn Asp Leu Tyr Arg Trp Glu Val Leu Ile Ile
30 35 40
GOC CCT CCA GAT ACA CTT TAT GAA GGT GGr GTT TTT AAG GCT CAT CTT 195
Gly Pro Pro Asp Thr Leu Tyr Glu Gly Gly Val Phe Lys Ala His Lau
45 50 55
AGT TTC OCA AAA GAT TAT COC CTC aGA CGT aCT AAF- ATG AAA TRC ATT 243
Thr Phe Pro Lys Asp Tyr Pro Leu Arg Pro Pro Lys Met Lys Phe Ile
60 65 70 75
ACA GAA ATC TGG CAC OCA AAT GTT GAT AAA AAT GGT GAT GTG TGC ATT 291
Thr Glu Ile Trp His Pro Asn Val Asp Lys Asn Gly Asp Val Cys Ile
80 85 90
TCT ATT CTT CAT GAG CCT GGG GAA GAT AAG TAT GGT TAT GAA AAG CCA 339
Ser Ile Leu His Glu Pro Gly Glu Asp Lys Tyr Gly Tyr Glu Lys Pro
95 100 105
GAG GAA OGC TGG CTC CCT ATC CAC ACT GTG GAA AOC ATC ATG ATT AGT 387
Glu Glu Arg Trp L8u Pro Ile His Thr Val Glu Thr Ile Met Ile Ser
110 115 120
GTC ATT TCT ATG CTG GCA GAC CGT AAT GGA GAC TCA CCT GGT AAT GTTT 435
Val Ile Ser Met Leu Ala Asp Pro Asn Gly Asp Ser Pro Ala Asn Val
125 130 135
CA 02458872 2004-03-23
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GAT GCT G0G AAA GAA TGG AOG GAA GAT AGA AAT GGA GAA TTT AAA AGA 483
Asp Ala Ala Lys Glu Trp Arg Glu Asp Ai+g Asn Gly Glu Phe Lys Arg
140 145 150 155
At,A GTT Gt7C GOC TGT GTA AGA AAA AOC CAA GAG AICT GCT TT'T GAG 528
Lys Val Ala Arg Cys Val Arg Lys Ser Gln Glu Thr Ala Phe Glu
160 165 170
TGACATITAT TTAGCAGGTA GTAACITCAC TTAT'iTCAGG GICTOf:AATT GAGAAAICATG 588
OC~T ~CZC~C~+CPC TAC0('.NJ(X` 617
(2) IMM,TION FOR SDQ ID NC): 25:
( i ) SEQUIIVCE CHARACTERIMICS:
(A) IE[VG't'fi: 374 aneino acids
( H ) TYPE: aomino ac3.d
(D) ToPOLOGY: linear
( ii ) NDIECXJLE TYPE: protein
( xi ) SEQUENCE DESQtIPTIaN: SDQ ID A10: 25 :
Met Val Leu Tip Glu Ser Pro Arg Gln Cys Ser Ser Trp Thr.LBu Cys
1 5 10 15
Glu Gly Phe Cys Trp Leu Leu Leu Leu Pro Val Met Leu Lau Ile Val
20 25 30
Ala Arg Pro Val Lys LSu Ala Ala Phe Pro Thr Ser L2u Ser Asp Cys
35 40 45
Gln Thr Pro Thr Gly Trp Asn Cys Ser Gly Tyr Asp Asp Arg Glu Asn
50 55 60
Asp Leu Phe Leu Cys Asp Thr Asn Thr Cys Lys Phe Asp Gly Glu Cys
65 70 75 80
L8u Arg Ile Gly Asp Thr Val Thr Cys Val Cys Gln Phe Lys Cys Asn
85 90 95
Asn Asp Tyr Val Pro Val Cys Gly Ser Asn Gly Glu Ser Tyr Gln Asn
100 105 110
Glu GYs Tyr Leu Arg Gln Ala Ala Cys Lys Gln Gln Ser Glu Ile Leu
115 120 125
Val Val Ser Glu Gly Ser Cys Ala Thr Asp Ala Gly Ser Gly Ser Gly
CA 02458872 2004-03-23
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130 135 140
Asp Gly Val His Glu Gly Ser Gly Glu Thr Ser Gln Lys Glu Thr Ser
145 150 155 160
Thr Cys Asp Ile Cys Gin Phe Gly Ala Glu Cys Asp Glu Asp Ala Glu
165 170 175
Asp Val Txp Cys Val Cys Asn Ile Asp Gys Ser Gin 'i'rr Asn Phe Asn
180 185 190
Pro Leu Cys Ala Ser Asp Gly Lys Ser Tyr Asp' Asn Ala Cys Gin Ile
195 200 205
Lys Glu Ala Ser Cys Gin Lys Gin Glu Lys.Ile Glu Val Met Ser Leu
210 215 220
Gly Arg Cys Gln Asp Asn Thr Thr Thr Thr Thr Lys Ser Glu Asp Gly
225 230 235 240
His Tyr Ala Arg Thr Asp Tyr Ala Glu Asn Ala Asn Lys Leu Glu Glu
245- 250 255
Ser Ala Arg Glu His His Ile Pro Cys Pro Glu His Tyr Asn Gly Phe
260 265 270
Cys Met His Gly Lys Cys Glu His Ser Ile Asn Met Gin Glu Pro Ser
275 280 285
Cys Arg Cys Asp Ala Gly Tyr Thr Gly Gln His Cys Glu Lys Lys Asp
290 295 300
Tyr Ser Val Leu Tyr Val Val Pro Gly Pro Val Arg Phe Gln Tyr Val
305 310 315 320
Iau Ile Ala Ala Val Ile Gly Thr Ile Gln Ile Ala Val Ile Cys Val
325 330 335
Val Val Leu Cys Ile Thr Arg Lys Cys Pro Arg Ser Asn Arg Ile His
340 345 350
Arg Gin Lys Gln Asn Thr Gly His Tyr Ser Ser Asp Asn Thr Thr Arg
355 360 365
Ala Ser Thr Arg Leu Ile
370
(2) INFORMATION FOR SEQ ID NO:26:
CA 02458872 2004-03-23
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(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1122 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:
ATGiC~I~CI~G'P 11OOO7 OCGOCRMM AO('.AOG'T+OG'A CACTTI'OfX'A OOOCt'1T1C~C 60
T+OOCt'OGRti~C TOCiCd00UGT CAZ=ACZC ATOOTAGt'70C GOI~(~OIG~~A GCTOOGTOCP 120
TTOOCPAOCP aCZTAAGIGA CTOOC,AkAOG QO(`.+~UOOOC'T Gx',AAT1CaC.'I~C TGG'iTAZG'AT
180
GACAGAGAAA ATGATCPCiT OC1C1C~iC`aAC AOCAAC,AOCP G!'AAATPiC',A ZTGOGGGAATOP 240
TTAAGAATTG GAGGiACAC7iUr GAL`Z't~00C~1PC ZC7J[~:A~C"A AQ'iGCAACAA TGAL'TATGRG
300
GCIC7CAATOG GGA~('aAOCTAC CAGkATGAGT GTPACCPOOG ACAGGGZCrCA 360
TGCAAAICAGC AGAGTGWAT ACTT.GTGC"1G TCAC'~AAiOGAT CAZGTOC7C".AC AGATGC'AGGA 420
TC;AIOGATCTG GAC'~AZGC''P OCATGAAOOC Tt:TGGAGAAA CTAGTCAAAA GGAGACATfJC 480
ACCTGhGA.TA TTZCrOCAGGPP TGGl'GCAGAA TGIC''~lOGAAG ATGOOGAGGA TC~1'CTOGIGT 540
=
GIGTOPAATA TT~GAC[GPTC 7~(`.AAA(7CAAC TTC',AATOCCC TC'iCXOGx.I'TC TGATOOGAAA
600
ZCTPATGATA AZnC'AZnOf'.A AATCAAAICiAA GCATCGI'Ol'C AGAAACAGGA GAAAATl'GAA 660
QPCATOTCIT TGOQiXX'aATG TCAAGATAAC ACAACTACAA CTAGTAAGTC TGAAC,ATGG(" 720
CATTAZGCAA GAACA[',ATTA ZGC'AGAGAAT aCCTTAAC'AAAT TAGAAGAAAG TGCCAGAGAA 780
CACCACATAIC CTl'G1b00C'aA iACATPACAAT GC~CI'TCIC~ICA TOCATGGGAA
G1GiC',AG1f',AT 840
ZCTATCAATA TGCA6iGi40CC ATCPiGCAGG TGZGAZC~C.`Z~G GITATACTGG ACAAClACTV'T 900
GAAAAAAAGG ACTACAQPOT ZC.TATAOGZT G7"I'OOOGiO1C CTOr1CGATT ZCAGTAZG'PC 960
TTAATOOC',AG CTQI'GATTOG AACAAT'ICAG ATTGCTGI'CA TCTGl'OTOGT GGi'OC'iC'MOC
1020
ATCACAAi+OGA AATGOOCCAG AAGCAACAGA ATPCACAIGAIC A-GAAOCAAAA TAIC,AGGOCAC 1080
TP,iCAGPTCAG ACAATACAAC AAGAGCGTCC ACGAI GTTAA TC 1122
CA 02458872 2004-03-23
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(2) INFORMATION FOR SEQ ID NO:27:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1721 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO.
(iv) ANTI-SENSE: NO
( vii ) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-092E10
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 368..1489
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:
~OG O~ QOCI~OCAOOC TGOCPOCi'CG GGCTOCI9C'I'C QI'CPGOOOT 60
GGACi'OOCGr CTCCTCCi'GT OCTO GGCTP COCAGAG= OCTOCITATG GCAGCAOCZT 120
COOGOGTCIC Q000G(',Ai(~CT TCIC'.AGOGGA OGAOOCIti`IC GCPOOGGGOC TC',AiGOCAG'TC
180
CCI'GC'aATGTT GC!'GAAACIC TCGAGp-TCAT GOCdOGGGI'iT GGCPOCPGCT TOOOOOC70GG 240
GTOOCAC'1GC CA~]CGOOOOC GOCPCi'GCIG UOGOOGIC)0(~ CGOC'aATGCPC AGTAGOOOGC 300
00G0C31kTaCT GPOr'IKACIM GAAGCOGTTT GC1GCPGCAG AGPIUCPdCGA 360
ACTA+GIC ATG GTG CTG TGG GAG TOC QOG aGG CAG TaC AGC AOC TOG ACA 409
Met Val Ieu Trp Glu Ser Pro Arg Gln Cys Ser Ser Trp Thr
1 5 10
CTT TGC GAG GGC TTT TOC TOG CTG CTG CTG GRG CCC GTC ATG CTA CTC 457
Lsu Cys Glu Gly Phe Cys Trp Leu Leu Lau Lau Pro Val Met Lau Leu
15 20 25 30
ATC GTA GCC CGC OOG GTG AAG CTC GCT GCP TTC CCT ACC TOC TTA AGT 505
Ile Val Ala Arg Pro Val Lys Leu Ala Ala Phe Pro Thr Ser Leu Ser
35 40 45
GAC TGC CAA ACG OCC AOC GGC TGG AAT TGC TCT GGT TAT GAT GAC AGA 553
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Asp Cys Gin Thr Prro Thr Gly Trp Asn Cys Ser Gly Tyr Asp Asp Arg
50 55 60
GAA AAT GAT CPC TTC CIC TGT GAC A,OC AAC AOC TGT AAA TTT GAT GGG 601
Glu Asn Asp Lau Phe Lsu Cys Asp Thr Asn Tthr Cys Lys Phe Asp Gly
65 70 75
GAA TGT TTA AGA ATT GGA GAC AGT GTG ACT TGC Gi'C TGT CAG TRC AP-G 649
Glu Cys Lsu Arg Ile Gly Asp Thr Val Thr Cys Val Cys Gln Phe Lys
80 = 85 90
TGC AAC AAT GAC TAT QIg aCT GTG TOT GGC TOC AAT GGG GAG AGC TAC 697
Cys Asn Asn Asp Tyr Val Pro Val Cys Gly Ser Asn Gly Glu Ser Tyr
95 100 105 110
CAG AAT GAG TGI' TAC = C!G OGA CAG GCT GCA TGC AAA CAG CAG AGT GAG 745
Gln Asn Glu Cys Tyr Leu Arg Gln Ala Ala. Cys Lys Gin Gln Ser Glu
115 120 125
ATA CLT G'M G't'G TCA GAA GGA TM TGT GOC ACA GAT GCA GGA TCA GGA 793
Ile Leu V.al Val Ser Giu Gly Ser Cys Ala Thr Asp Ala Gly Ser Gly
130 135 140
Tt,'T GGA. GAT GGA GTC CAT GAA GGC TL:T GGA GAA ACT AGT CAA AAG GAG 841
Ser Gly Asp Gly Val His Glu Gly Ser Gly Glu Ztr Ser Gin Lys Glu
145 150 155
ACA TOC AOC TGT GAT ATT = CAG TTT GGT GCA GAA TGT GAC GAA GAT 889.
Thr Ser Thr Cys Asp Ile Cys Gln Phe Gly Ala Glu Cys Asp Glu Asp
160 165 170
GOC GAG GAT GTC TGG TGr G1'G TGT AAT ATT GAC TOT RCT CAA AOC AAC 937
Ala Glu Asp Val Trp Cys Val Cys Asn Ile Asp Cys Ser Gin Thr Asn
175 180 185 190
TTC AAT COC CPC TGC GCT TCT GAT GOG AAA TGT TAT GAT AAT GCA TGC 985
Phe Asn Pro Leu Cys Ala Ser Asp Gly Lys Ser Tyr Asp Asn Ala Cys
195 200 205
CAA ATC AAA GAA GCA TOG TGT CAG AM CAG GAG AAA ATT GAA GIC ATG 1033
Gln Ile Lys Glu Ala Ser Cys Gln Lys Gln Glu Lys Ile Glu Val Met
210 215 220
TCT TTG GGT GGA TGT CAA GF-T AAC ACA AiCT ACA ALT ACT APuG TCT GAA 1081
Ser Leu Gly Arg Cys Gin Asp Asn Thr Thr Thr Tthr Thr Lys Ser Glu
225 . 230 235
GAT GGG CAT TAT GCA AGA ACA GAT TAT GCA GAG AAT Gt:T AAG AAA TTA 1129
Asp Gly His Tyr Ala Axg Thr Asp Tyr Ala Glu Asn Ala Asn Lys Leu
240 245 250
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GAA GAA AGT GOC AGA GAA CAC CAC ATA OCT TGT C70G GAA CAT TAC AAT 1177
Glu Glu Ser Ala Arg Glu His His Ile Pro Cys Pro Glu His Tyr Asn
255 260 265 270
GGC TZC TGC ATG CAT GGG AAG TGT GAG CAT TCT ATC AAT ATG CAG GAG 1225
Gly Phe Cys Met His Gly Lys Cys Glu His Ser Ile Asn Met Gln Glu
275 280 285
CCA TGT TGC AGG TGT GAT GCT GOT TAT AGT OGA CAA CAC TGT GAA AAA 1273
Pro Ser Cys Arg Cys Asp Ala Gly Tyr Thr Gly Gln His Cys Glu Lys
290 295 300
AAIGGACTACAO,'TGRTGTATACG'1TGTTOCCG(GPOCTGTAOGATTTCAG 1321
Lys Asp Tyr Ser Val Lau Tyr Val Val. Pro Gly Pro Val Arg Phe Gln
305 310 315
TAT GTC TTA ATC G1C;A GCT GIG ATT GGA ACA ATT CAG ATT GCT GTC ATC 1369
Tyr Val. Leu Ile Ala Ala Val Ile Gly Thr Ile Gln Ile Ala Val ile
320 325 330
Tt7r GTG GTG GTC CTC TGC ATC ACA AGG AAA TGCC COC AGA AOC AAC AGA 1417
Cys Val Val Val L8u Cys Ile Thr Arg Lys Cys Pro Arg Ser Asn Arg
335 340 345 350
ATT CAC AGA CAG AAG CAA AAT ACA GOG CAC TAC AGT TGA GAC AAT ACA 1465
Ile His Arg Gln Lys Gin Asn Thr Gly His Tyr Ser Ser Asp Asn Thr
355 360 365
ACA AGA GOG TaC AIOG AOG TPA ATC TAA AGGGAGCATG TTTCACAGM 1512
Thr Arg Ala Ser Thr Arg Leu Ile
370
GCICaGACl'AC CGAGAG7CI"i'G GGACTACAC'AA TACIAG'PATTA TAGACAAAAG AATAA~'aAIGAA
1572
GAG'ATG'TACA CATGTIGOCP T(~('ATITQIG GTAATCTACA OCAAZCAAAA CATMACPAC 1632
,AiOCTATATZT GATTAZGPAT GGATATATZT GAAATAQTAT ACATTGiti`iT GATGPT'IRTP 1692
CTGPAATGPA AATAAAGTAT TTATATCAC 1721
(2) INFORNATION FOR SnQ ID ND:28:
(1) SEQUENCE t3RARAC'PEZISTICS :
(A) LENGtH: 817 aiaino acids
( B ) TYPE: amino acid
( D ) TOPOIAG'Y: linear
( ii ) NlOLECxTLE TYPE: protein
CA 02458872 2004-03-23
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( xi. ) SDQUIIViGE DFSCRIPTION: SHQ ID NO: 28:
Met Gly Asp Thr Val Val Glu Pro Ala Pro Leu Lys Pro Thr Ser Glu
1 5 10 15
Pro Thr Ser Gly Pro Pro Gly Asn Asn Gly Gly Ser Leu Leu Ser Val
20 25 30
Ile Thr Glu Gly Val. Gily Glu Leu Ser Val Ile Asp Pro Glu Val. Ala
35 40 45
G]n Lys.Ala Cys Gln Glu Val Leu Glu Lys Val Lys Leu Lieu His Gly
50 55 60
Gly Val Ala Val Ser Ser Arg Gly Thr Pro Leu Glu Leu Val Asi Gly
65 70 75 80
Asp Gly Va1 Asp Ser Glu Ile Arg Cys Leu Asp Asp Pro Pro Ala Gln
85 90 95
Ile Arg Glu Glu Glu Asp Glu Met Gly Ala Ala Val Ala Ser Gly Thr
100 105 110
Ala Lys Gly Ala Arg Arg Arg Arg Gln Asn Asn Ser Ala Lys Gln Ser
115 120 125
Txp Leu Lau Arg leu Phe Glu Ser Lys Loau Phe Asp Ile Ser Met Ala
130 135 140
Ile Ser Tyr Leu Zyr Asn Ser Lys Glu Pro Gly Val Gln Ala Tyr -Ile
145 150 155 160
Gly Asn Arg Leu Phe Cys Phe Arg Asn Giu Asp Val Asp Phe Tyr Leu
165 170 175
Pro Gin Leu Leu Asn Met Tyr Ile His Met Asp Glu Asp Val Gly Asp
180 185 190
Ala Ile Lys Pro T-yr Ile Val His Arg Cys Arg Gln Ser Ile Asn Phe
195 200 205
Ser Leu Gln Cys Ala Leu L8u Leu Gly Ala Tyr Ser Ser Asp Met His
210 215 220
Ile Ser Thr Gin Arg His Ser Arg Gly Thr Lys Leu Arg Lys Leu Ile
225 230 235 240
Leu Ser Asp Glu Leu Lys Pro Ala His Arg Lys Arg Glu Leu Pro Ser
245 250 255
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Leu Ser Pm Ala Pro Asp Thr Gly Leu Ser Pro Ser Lys Arg Thr His
260 265 270
Gln Arg Ser Lys Ser Asp Ala Thr Ala Ser Ile Ser Leu Ser Ser Asn
275 280 285
Leu Lys Arg Thr Ala Ser Asn Pro Lys Val Glu Asn Glu Asp Glu Glu
290 295 300
Leu Ser Ser Ser Ttvc Glu Ser Ile Asp Asn Ser Phe Ser Ser Pro Val
305 310 315 320
Arg Leu Ala Pro Glu Arg Giu Phe Ile Lys Ser Lau Met Ala Ile Gly
325 330 335
Lys Arg Leu Ala.Thr Leu Pro Thr Lys Glu G]n Lys'Thr Gin Arg Leu
340 345 350
Ile Ser Glu Leu Ser Leu L$u Asn His Lys LSu Pro Ala Arg Val Trp
355 360 365
LSu Pro Thr Ala Gly Phe Asp His His Val Val Arg Val Pro His Thr
370 375 380
Gin Ala Val Val Leu Asn Ser Lys Asp Lys Ala Pro Tyr Leu Ile Tyr
385 390 395 400
Val Glu Val Leu Glu Cys Glu Asn Phe Asp Thr Thr Ser Val Pro Ala
405 410 415
Arg Ile Pro Glu Asn Arg Ile Arg Ser Thr Arg Ser Val Glu Asn Leu
420 425 430
Pro Glu Cys Gly Ile Thr His Glu Gln Arg Ala Gly Ser Phe Ser Thr
435 440 445
Val Pro Asn Tyr Asp Asn Asp Asp Glu Ala Trp Ser Val Asp Asp Ile
450 455 460
Gly Glu Leu Gln Val Glu Leu Pro Glu Val His Thr As.n Ser Cys Asp
465 470 475 480
Asn Ile Ser Gln Phe Ser Val Asp Ser Ile Thr Ser Gln Glu Ser Lys
485 490 495
Glu Pro Val Phe Ile Ala Ala Gly Asp Ile Arg Arg Arg Leu Ser Glu
500 505 510
Gin Leu Ala His Thr Pro Thr Ala Phe Lys Arg Asp Pro Glu Asp Pro
515 520 525
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Ser Ala Val Ala Leu Lys Glu Prcm Trp Gin Glu Lys Val Arg Arg Ile
530 535 540
Arg Glu Gly Ser Pro Tyr Gly His Leu Pro Asn Trp Arg Leu Leu Ser
545 550 555 560
Val Ile Val Lys Cys Gly Asp Asp Leu Arg GIn Glu Leu Leu Ala Phe
565 570 575
Gin Val Leu Lys Gln Leu Gln Ser Ile Trp Glu Gin Glu Arg Val Pro
580 585 590
Leu Trp Ile Lys Pro Ile Gln Asp Ser Cys Glu Ile Thr Thr Asp Ser
595 600 605
Gly Met Ile Glu Pro Val Val Asi Ala Val Ser Ile His Gln Val Lys
610 615 620
Lys Gin Ser Gln Leu Ser LSu Leu Asp Tyr Phe Lau Gln Glu His Gly
625 630 635 640
Ser Tyr Thr Thr Glu Ala Phe Leu Ser Ala Gln Arg Asn Phe Val Gln
645 650 655
Ser Cys Ala Gly Tyr Cys Leu Val Cys Tyr Leu Leu Gin Val Lys Asp
660 665 670
Arg His Asn Gly Asn.Ile Lau Leu Asp Ala Glu Gly His Ile Ile His
675 680 685
Ile Asp Phe Gly Phe Ile Leu Ser Ser Ser Pro Arg Asn Leu Gly Phe
690 695 700
Glu Ttir Ser.Ala Phe Lys Leu Thr Thr Glu Phe Val Asp Val Met Gly
705 710 715 720
Gly Leu Asp Gly Asp Met Phe Asn Tyr Tyr Lys Met Leu Met Leu Gin
725 730 735
Gly Leu Ile Ala Ala Arg Lys His Met Asp Lys Val Val Gin Ile Val
740 745 750
Glu Ile Met Gln Gln Gly Ser Gln Leu Pro Cys Phe His Giy Ser Ser
755 760 765
Thr Ile Arg Asn Leu Lys Glu Arg Phe His Met Ser Met Thr Glu Glu
770 775 780
Gln Leu Gln Leu L2u Val Glu Gln Met Val Asp Gly'Ser Met Arg Ser
785 790 795 800
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Ile Thr Thr Lys Leu Tyr Asp Gly Phe Gln Tyr LQu Thr Asn Gly Ile
805 810 815
Met
(2) INFORMATION FOR SEQ ID NO:29:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2451 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE'DESCRIPTION: SEQ ID NO:29:
AZGDf',,,AGATA CAGTAOrGC'aA GCCTGOC0(7C TPGAAGOCAA CPPCTC'aA00C CACTTCI'GOC 60
CCACCAGGGA ATAAZUOGOG GTUCGRC~C'rA AGPG'ir'ATCA. CGC"~GGGGGT aGGGGAACTA 120
TCAG'IC'aATTG AICCCTGAGGT GGOCC'AGAAG GOCIGOCAGG AOGI'GrTGGA GAAAGTCAAG 180
CTZTIGCATG GAGGOGTGGC AGTCrCrAGC AGAGGCAIOCC CACiGUAIT GGZC'.AATGGG 240
GATOGTGTGG ACAG-IGAGAT CC7Gr'IGCCrA GATGATCCAC C'I'GCCCAGAT CAGGGAGC'~FUG 300
GAA!('~ATGAiGA TOGGGOOOOC TG'rGOC7C'YCA GGCAC,l1O0CA AAGGAGt'AAG AAGAlUGGOGG
360
CAGAACAACr CAGC.TAAACA GICTrCG~COC.TG CTGAGGCTGT 420
ATC.'rOCATGG CJCARTTCATA CCIGTATAAC TaC'AAGCAGC C.IC'GAGTACA AOC7CTACATT 480
GGCAACCGGC TCT'I'CTCM ZbGCAAOG?1G GACGPQGACT Tr7rATCPC~fJC OCAGITGCTT 540
AACATG'rACA TCCACATGGA TGAGGAOGPG GGI'GATGCCA TTAAGOO~,.TA CATAGTCCAC 600
CG'r'P00MCC AGAGC'ATTAA C'I'TTTCCCTC CAGTGI'GCCC TGTZGCTrGG GGCCrATiCT 660
TCAGACATGC ACATr'I'C7C`,AC TCAAOGACAC TCCCGrGGGA CCAAGC,TACG GAAGC,'TC',ATC
720
C'1C.'TCAGATG AGCTAAAGOC AGCTCAC'.AGG AAGAGOGAOC TGOOCTOCPT GAGOCCGGCC 780
CC.'TGATACAG GGCIGTCI'CC C'POCAAAAGG ACZC'ACCAGC GC.ZCTAAG'IC AGATGCCACT 840
G(7CAGC'ATAA GTCTCAGCAG C'AACCTGAAA CGAACAGCCA pCAAOnCrAA AGTGGAGAAT 900
GAGGATGAGG A+CGCTG=rCCTC Cl4GCACOGAG AGTATTGATA ATTCATI'CAC TTOCCCTGTT 960
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C7C'~ACTGGC,'TC CTGAGAGAGA ATTCATCAAG TCxCZGATGG CGAT00GC'.AA GIXGGC'TGOOC
1020
AOGCTOOOCA OC'AAAGAGCA GAAAAICACAG AGGCTGF-TCT CAGAGCTG'rC C7CI'GCTCAAC 1080
CATAAGCTCC C.TGOCXX"'aAGT CTGGGTGOCC AC.TIX7rGGCT TTGAi0CA0CA OGrOGrQO(~`r
1140
GPAOOOCAICA CAICAGGCTGT TGrC:C'ICAAC TC)CAAGGAlC'.A AGGCr00CrA OCrGATTTAT 1200
GTGf",AAGrCC TIGAATGTGA AAAIC'IRT(`lIC AOCAOCAGZG TCJOC.TGO(70G GAZ"OOOOGAG
1260
AAC70GAP,TTC GGAGTACGAG Gr000PACAA AACTTGOOCG AATGrGGTAT TAOOC'.ATGAG 1320
CAGOGAQCTG GC'AOCIrCAG CACTGr000C AACTA4G~AC'A AOC'1TGA7GA GGOC'IC~GTCG 1380
GTGGATGACA TAGGCJGAGCr GCAAG'rGGAG C'r000('aAA[" TGCATAIX'.AA CAGGPGTGFUC 1440
AAICATCr= AGTZCTCWr GGAiCAGC''ATC A+OCAG(7C',AGC' AGAOC'.AAOGA GOC'IC~I'GR'rC
1500
ATTGCAOC'AG CiGGAC'ATOCIG ~7~OOOOCTr TCGCGAAC,AG'C 'rGGCTC'ATAC OOOGACAGOC
1560
TTCAAACGAIG AOOCA~'~AAGA TCGRTC.TGC'A GTTGCTCrCA AAGAGOOCrG GCõAQGAGAAA 1620
GTACGGOGGA TCAGAGAGGG CTOOOOCTAC GGOCATC'IC7C OC.AATIG(.aOG GC1Y+~OCrGrCA 1680
GrCATZGTCA AIGPGI`GGGGA TGAOC`IRO~G CAFIIC'zP,IOCPTC TGOOG'ZTIC'A GGPGPTGeAAG
1740
CAACrGCAQr CCATT'PGGGA. AC,AGGAGOf3A GrO000C'PIT GGATC.AAOOC AATACAP-GAT 1800
TCTTGIGAAA TTAOGAICTGA TAGTGGCATG ATTGAAOCAG TGGTC'AAT C TGRGTCC'ATC 1860
CATC'.AOC=rGA AIGAAAC.AGrC ACArGC'I'CTOC TTGCTC+GATT AL'TTOCTACA GGAGCAOGaC
1920
AGTTACAC7CA CPGAGGiC'.ATT C1C:TCAGrOCA CAGCGCAATT TI"GTGCAAAG TTG~`GC'IGGG
1980
TAGTGiC,'TTGG TCTGC.`rAOCT GCTaCAAGTC AAGGACACdAC ACAATGGGAA TATOG'iTTTG 2040
GACGCAGAAG GOCACP-TCAT OCACATO(3AC TTTQGCTrCA TOCTCiCX'.AG CrCA,0000GA 2100
AATC:TCGGCr TTGAGACGrC AGOCT'rrAAG cTC3AloCAC.AG AGTTrGI'GGA TGc1GATGGGC 2160
aGOC'rGGPiTG GOGACATGZT CAACrAGTAT AAGATGCrGA TGCTGC'AAOG GCZGATTGOC 2220
aC'ICM~ ACATGGAMA GG'rOG'rOCAIG ATOG'1GGAGA TCATaCAGC'A AQGITCTCAG 2280
CTTaCTTGCT TC7CATGIX,`TC CAGCAOC'ATT OGAAAOCPCA AFuGAGAGGTr OCACATGAGC 2340
ATGF,CrGADG AGCAGCIGCA GG'ZGCrGGrG GAGC'AGP-ZGG TGGATOOCAG TATGO(YG'rCT 2400
ATCAQC.ACCA AACTCTATGA c7GGCTI'OCAG TAOCTCAOCA ACGa('.ATCAT G 2451
CA 02458872 2004-03-23
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(2) INFORMATION FOR SEQ ID NO:30:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3602 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-428B12c2
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 429..2879
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:
GGTGGCTCAC GCCPGPAATC CCAGCAC.`ITT CaGGAGGoACAA GCAGATOfJC TTCAOOOCAG 60
GAGGTAGAGG CTGCAG'TGAG CPGI'~,~ATGGP GC(',ACTGC'.AC TCCAGCC'IM GCAATGAAGC 120
AAGA=AT CTC%kkkkkkJk AAATITTPAA AAAAGGCAAA GATGGGCX.TG GGGCACCAAA 180
TAZTCCI4GAG GAAAGGGAAC G'PGlGTACTC CTTGAGGTOG GGAACATGAC (7CACT'PGAGG 240
TGC',AGAAAGA AGAC.'TTGTAT GGGGC'I'GGTG CAGCCTCOGC GGOOGCTGIC AOGC'~AAOCGC 300
AQGOOGCCAA TGGAACCOGG GAGCGG'i'CGC ZGCTO(.~TGAG GOGGCAGTGP CGGC',AG'rCCA 360
AOCOC7GACIG CCCGCAOCCC C'POOOCGGGIG TCOOOCAGAG CTIGC'aAAGCP CGkAGTCTGG 420
Ci'GM3GCC ATG GGA GAT ACA GTA GTG GAG CCT OC CCC TTG AAG CCA ACT 470
Met Gly Asp Thr Val Val Glu Pro Ala Pro Leu Lys Pro Thr
1 5 10
TCT GAG aCC ACT TCT GOC CCA CCA GGG AAT AAT GGG GGG TOC CTG CTA 518
Ser Glu Pro Thr Ser Gly Pro Pro Gly Asn Asn Gly Gly Ser Leu Leu
15 20 25 30
AGT GTC ATC ACG GAG GGG GTC GGG GAA CTA TCA GTG ATT GAC CCT GAG 566
Ser Val Ile Thr Glu Gly Val Gly Glu Leu Ser Val Ile Asp Pro Glu
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35 40 45
GTG GOC CAG AAG GC1C TGC CAG GAG GTG TTG GAG AAA GTC AAG CI'T TTG 614
Val Ala Gln Lys Ala Cys Gln Glu Val LSu Glu Lys Val Lys L8u Leu
50 55 60
CAT GGA GOC GTG GCA GTC TGT AGC AGA GGC AOC CCA CTG GAG T'IG G'TC 662
His Gly Gly Val Ala Val Ser Ser Arg Gly Thr Pro Leu Glu L2u Val
65 70 75
AAT GOG GAT GGT GTG GAC AGT GAG ATC CGr TGC G'TA GAT GAT aC'A CCT 710
Asn Gly Asp Gly Val Asp Ser Glu Ile Arg Cys Leu Asp Asp Pro Pro
80 85 90
GUC CAG ATC AGG GAG GAG GAA GAT GAG ATG GOG GCC GCT GTG GOC TCA 758
Ala Gln Ile Arg Glu Glu Glu Asp Glu Met Gly Ala Ala Val Ala Ser
95 100 105 110
GOC ACA GOC AAA GGA GCA AGA AGA CJGG COG CAG AAC AAC TCA GC,'T AAA 806
Gly Thr Ala Lys Gly Ala Arg Arg Arg Arg Gln Asn Asn Ser Ala Lys
115 120 125
CAG TCT TaG CTG CTG AOG C.TG TTT GAG TCA AAA CZG TTT GAC ATC TOC 854
Gln Ser Trp Leu Leu Arg Leu Phe Glu Ser Lys Leu Phe Asp Ile Ser
130 135 140
ATG GCC ATT TCA TAC CTG TAT AAC TCC AAG GAG CCT GGA.GTA CAA GOC 902
Met Ala I1e Ser Tyr Leu Tyr Asn Ser Lys Glu Pro Gly Val Gin Ala
145 150 155
TAC ATT GGC A24C CX1G CIC TTC TGC TZT CGC AA+C GAG GAC GI'G GAC TTC 950
Tyr Ile Gly Asn Arg Leu Phe Cys Phe Arg Asn Glu Asp Val Asp Phe
160 165 170
TAT CTG OC;C CAG TTG C1T AAC ATG TAC ATC CAC ATG GAT GAG GAC GTG 998
Tyr Leu Pro Gln Lsu Leu Asn Met Tyr Ile His Met Asp Glu Asp Val
175 180 185 190
GGT GAT GCC ATT AAG CCC TAC ATA GTC CAC CGT TGC CGC CAG AGC AZT 1046
Gly Asp Ala Ile Lys Pro Tyr Ile Val His Arg Cys Arg Gln Ser Ile
195 200 205
AAC TTT TCC CTC CAG TGT GOC CTG TTG CTT GflG GOC TAT TLT TCA GAC 1094
Asn Phe Ser Leu Gln Cys Ala Leu Leu Leu Gly Ala Tyr Ser Ser Asp
210 215 220
ATG CAC ATT TOC ACT CAA CGA CAC TCC aG=I' GGG ACC AAG CTA aGG AAG 1142
Met His Ile Ser Thr Gln Arg His Ser Arg Gly Thr Lys Lsu Arg Lys
225 230 235
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CPG ATC CIC TCA GAT GAG CTA AAG CCA GCT CAC AGG AAG AGG GAG CiG 1190
Leu Ile Leu Ser Asp Glu Leu Lys Pro Ala His Arg Lys Arg Glu Lsu
240 245 250
COC TCC TTG AIDC OOG GOC OCT GAT ACA GGG CTG TCT UOC TOC AAA AGG 1238
Pro Ser Lau Ser Pro Ala Pro Asp Thr Gly Leu Ser Pro Ser Lys Arg
255 260 265 270
ACT CAC CAG OGC TGZ' AAG TCA GAT GOC ACT GrJC AOC ATA AGT CTC AOC 1286
Thr His Gln Arg Ser Lys Ser Asp Ala Thr Ala Ser Ile Ser Leu Ser
275 280 285
AGC AAC CI'G AAA OGA ACA GGC AGC AAC OC't' AAA GTG GAG AAT -GAG GAT 1334
Ser Asn Leu Lys Arg Thr Ala Ser Asn Pro Lys Val Glu Asn Glu Asp
290 295 300
GAG GAG CrC TCC TrJC,AGC ACC GAG AGT ATT GAT AAT TCA TTC AGT TaC 1382
Glu Glu Leu Ser Ser Ser Thr Glu Ser Ile Asp Asn Ser Phe Ser Ser
305 310 315
(7CT GTT OGA C'IG GCT CCT GAG AGA GAA TTC ATC AAG TOC CPG ATG GOG 1430
Pro Val Arg Leu Ala Pro Glu Arg Glu Phe Ile Lys Ser Leu Met Ala
320 325 330
ATG GOC AAG CJGG C1G GOC AfJG GTC QOC ACC AAA GAG CAG AAA ACA CAG 1478
Ile Gly Lys Arg Leu Ala Thr Leu Pro Thr Lys Glu Gln Lys Thr Gln
335 340 345 350
AGG C'i'G ATC TCA GAG C'I'C TCC CIG CTC AAC CAT AAG C'IC CCT GC7C CGA 1526
Arg Lsu Ile Ser Glu Leu Ser Lau Leu Asn His Lys Leu Pro Ala Arg
355 360 365
GTC TOG C'I'G Ot7C ACT GCT GGC T'IT GAC CAC CAC GTG GTC C)GT GTA OOC 1574
Val Tzp Leu Pro Thr Ala Gly Phe Asp His His Val Val Arg Val Pro
370 375 380
CAC ACA CAG GCT G'I'T GTC CTC AAC TOC AAG GAC AAG GCT UOC TAC CtG 1622
His Thr Gln Ala Val Val Leu Asn Ser Lys Asp Lys Ala Pro Tyr Leu
385 390 395
ATT TAT GIG GAA G'TC CTT GAA TGT GAA AAC TTT GAC ACC ACC AGT GTC 1670
Ile Tyr Va]. Glu Val Leu Glu Cys Glu Asn Phe Asp Thr Thr Ser Val
400 405 410
CCT GOC CAG ATC COC GAG AAC CGA ATT CGG AGP AOG AGG TOC GTA GAA 1718
Pro Ala Arg Ile Pro Glu Asn Arg Ile Arg Ser Thr Arg Ser Val Glu
415 420 425 430
AAIC TTG C:OC GAA TGT GGT ATT AGC CAT GAG CAG CGA GCT GflC AGC TTC 1766
Asn Leu Pro Glu Cys Gly Ile Thr His Glu Gln Arg Ala Gly Ser Phe
CA 02458872 2004-03-23
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435 440 445
AGC ACT GTG COC AAC TAT GAC AAC GAT GAT GAG GOC TGG TCG GTG GAT 1814
Ser Thr Val Pro Asn Tyr Asp Asn Asp Asp Glu Ala Trp Ser Val Asp
450 455 460
GAC ATA GGC GAG CTG CAA GTG GAG C'I'C COC GAA GIG CAT AOC AAC AGC 1862
Asp Ile Gly Glu Leu Gin Val Glu Leu Pro Glu Val His Thr Asn Ser
465 470 475
TGP GAC AAC ATC TCC t'AG TTC TGT GTG GAC AGC ATC AOC AGC CAG GAG 1910
Cys Asp Asn Ile Ser Gln Phe Ser Val Asp Ser Ile Thr Ser Gin Glu -
480 485 490
AOC AAG GAG OCT G'I'G Tl'C ATT OCA GCA GGG GAC ATC OGC (]OG COC CIT 1958
Ser Lys Glu Pro Val Phe Ile Ala Ala Gly Asp Ile Arg Arg Arg Leu
495 500 505 510
TOG GAA CAG CTG GCT CAT ACC OOG ACA GOC TIC AAA OGA GAC aCA GAA 2006
Ser Glu Gin Leu Ala His Thr Pro Thr Ala Phe Lys Arg Asp Pro Glu
515 520 525
GAT (X.T TC.T GCA GTT GCT CIC AAA GAG OOC TOG CAG GAG AAA GTA C7GG 2054
Asp Pro Ser Ala Val Ala Leu Lys Glu Pro Trp Gin Glu Lys Val Arg
530 535 540
CGG AZC AGA GAG GGC TOC C70C TAC GGC CAT CTC COC AAT ZGG CGG GTC 2102
Arg Ile Arg Glu Gly Ser Pro Tyr Gly His Leu Pro Asn Trp Arg Leu
545 550 555
C'IG 'I'CA GTC ATT GTC AAG TGT GGG GAT GAC CTT OOG CAA GAG CIT CPG 2150
Leu Ser Val Ile Val Lys Cys Gly Asp Asp Leu Arg Gln Glu Leu LSu
560 565 570
GCC TTT CAG GTG TTG AAG CAA CTG C14G TOC ATT TOG GAA CAG GAG OGA 2198
Ala Phe Gin Val Leu Lys Gln Leu Gin Ser Ile Trp Glu Gin Glu Arg
575 580 585 590
GTG COC CTT TGG ATC AAG C3CA ATA CAA GAT TCT TGT GAA AZT AOG ACT 2246
Val Pro Leu Tip Ile Lys Pro Ile Gln Asp Ser Cys Glu Ile Thr Thr
595 600 605
GAT AGP GOC ATG AZT GAA aCA GTG GTC AAT GCT GIG TOC ATC CAT C'AG 2294
Asp Ser Gly Met Ile Glu Pro Val Val Asn Ala Val Ser I1e His Gln
610 615 620
GTG AAG RAA CAG TCA CAG CTC TC7C TTG CTC GAT TAC 'ITC CTA CAG GAG 2342
Val Lys Lys Gln Ser Gln Leu Ser Leu Leu Asp Tyr Phe Leu Gln Glu
625 630 635
CA 02458872 2004-03-23
-152-
CAC GGC AGT TAC AOC ACT GAG GCA TTC CTC PM GCA CAG OOC AAT m 2390
His Gly Ser Tyr Thr Thr Glu Ala Phe Lau Ser Ala Gln Arg Asn Phe
640 645 650
GTG CAA AGT TGP GC,'T GGG TAC TGC TTG GTC TGC TAC CTG CTG CAA GTC 2438
Val Gln Ser Cys Ala Gly Tyr Cys Leu Val Cys Tyr Leu Leu Gln Val
655 660 665 670
AAG GAC AGA CAC AAT GGG AAT ATC CTT TTG GAC GCA GAA GGC CAC ATC 2486
Lys Asp Arg His Asn Gly Asn Ile Leu Leu Asp Ala Glu Gly His Ile
675 680 685
ATC CAC ATG GAC TTT GOC TTC ATC CrC TOC AGC TCA OOC aGA AAT CTG 2534
Ile His Ile Asp Phe Gly Phe Ile Leu Ser Ser Ser Pro Arg Asn Leu
690 695 700
GGC TiT GAG AOG TGF- GOC TTT AAG CTG ACC ACA GAG TTT GTG GAT GTG 2582
Gly Phe Glu Thr Ser Ala Phe Lys Leu Thr Thr Glu Phe Val Asp Val
705 710 715
ATG GGC GGC CTG GAT GGC GAC ATG TTC AAC TAC TAT AAG ATG CTG ATG 2630
Met Gly Gly Leu Asp Gly Asp Met Phe Asn Tyr Tyr Lys Met Leu Met
720 725 730
C'i'G CAA GGG CTG ATT GCC GL'T CJGG AAA CAC ATG GAC AAG GTG GTG CAG 2678
Leu Gln Gly Leu Ile Ala Ala Arg Lys His Met Asp Lys Val Val Gln
735 740 745 750
ATC GTG GAG ATC ATG CAG CAA GGT TCT CAG CTT aCT TOC TTC CAT GGC 2726
Ile Val Glu Ile Met Gin Gln Gly Ser Gln Leu Pro Cys Phe His Gly
755 760 765
TOC AGC ACC ATT CGA, AAIC CTC AAA GAG AGG TTC CAC ATG AGC ATG ACT 2774
Ser Ser Thr Ile Arg Asn Leu Lys Glu Arg Phe His Met Ser Met Thr
770 775 780
GAG GAG CAG CTG CAG CTG CTG GTG GAG CAG ATG GZG GAT GGC AGT ATG 2822
Glu Glu Gln Leu Gln Leu Leu Val Glu G1n Met Val Asp Gly Ser Met
785 790 795
COGG TCT ATG ACC ACC AAA CI'C TAT GAC GGC TTC CAG TAC CTC ACC AAC 2870
Arg Ser Ile Thr Thr Lys Leu Tyr Asp Gly Phe Gln Tyr Leu Thr Asn
800 805 810
GGC ATC ATG TGA CA(JGCTOCri'C AGOQCAGGAG 'PGGTGGGGGGG TaGAGGGCAC 2922
Gly Ile Met *
815
(7CTOOC,'TAIGA uAGOCC,`PTGT CTGAGAAACC aC'AAAOCAGG AAACOOCA<7C TAOOCAP-CCA
2982
CA 02458872 2004-03-23
-153-
TOC.ACOCAAG GGAAAZGGAA CG('.AAGAAAC AOGAAGGATC AZGIXMAAC TGOGAGAGCT 3042
TaCTGlUOGGG TGGC'aAC",~AGC1C AGCTGi'OGGG TCCAGACI=I'G ZTGGC~C'ITC
CC'IGCOCC.'TC 3102
CTGG!'C'IGI'G TCAGTAZTAC CACC.A~f`aA 'IG AC'TCCAGGAC TCACTGCCCT OCAGAAAACA
3162
GAD(7rGACAA ATGtr-%AGGGA CAC'1'GGOGCC TTTCITG`I'CC TTGTAGGGGP CZCTCAGAOG 3222
T'iC'ITTaCAC AGGCCATCCT CTTATTC7W'T TC.Z'GGGGOOC AGGAAGTGGG GAAGAGTAGG 3282
TTC'I'C1G(OPAC TTAGGACT'iG ATCCTG'IWT TGCCACTGGC CATGCI'GCTG CCCAGCTCPA 3342
OOCC'iC70C'.AL'' Gf'~40CTACOC C'i"CCC.AOGGA COGACCOC't'G GCCCAAGCTC OOLTAGCIGG
3402
OGOGCGC:ZGC G'POOOOUCIG CACTi'GCTC'~+, GG'l.'ZC700C'AT CATGGGCAAG GCAAGGGAAT
3462
Tl7CCACAGCC CTCCAGTGTA CTGAOdGPAC TGGOG'I'AGCC ATO!'GGAATT CCCTACO~.~TG 3522
ACI'OC`iTCCC CAAAC70CAGG GAAAAGAGC.T C'trAAT'ITPP TAZTPITAAT ZTTTGITI3582
AATAAAGIC7C TTAG'TPAGCC 3602
(2) INFORMATION FOR SEQ ID NO:31:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 829 amino acids
(B) TYPE: amino acid
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:
Met Arg Phe Leu Glu Ala Arg Ser Leu Ala Val Ala Met Gly Asp Thr
1 5 10 15
Val Val Glu Pro Ala Pro Leu Lys Pro Thr Ser Glu Pro Thr Ser Gly
20 25 30
Pro Pro Gly Asn Asn Gly Gly Ser Leu Leu Ser Val. Ile Thr Glu Gly
35 40 45
Val Gly Glu Leu Ser Val Ile Asp Pro Glu Val Ala Gin Lys Ala Cys
50 55 60
Gln Glu Val Leu Glu Lys Val Lys Leu Leu His Gly Gly Val Ala Val
65 70 75 80
CA 02458872 2004-03-23
-154-
Ser Ser Arg Gly Thr Pro Lau Glu Leu Val Asn Gly Asp Gly Val Asp
85 90 95
Ser Glu Ile Arg Cys Leu Asp Asp Pro Pro Ala Gin Ile Arg Glu Glu
100 105 110
Glu Asp Glu Met Gly Ala Ala Val Ala Ser Gly Thr Ala Lys Gly Ala
115 120 125
Arg Arg Arg Arg Gin.Asn Asn Ser Ala Lys Gin Ser Trp Leu Leu Arg
130 135 140
Leu PYye Glu Ser Lys Leu Phe Asp Ile Ser Met Ala Ile Ser Tyr Leu
145 150 155 160
Tyr Asn Ser Lys Glu Pro Gly Val Gln Ala Tyr Ile Gly Asn Arg Leu
165 170 175
Phe Cys Phe Arg Asn Glu Asp Val Asp Phe Tyr Leu Pro Gln Isu Leu
180 185 190
Asn Met Tyr Ile His Met Asp Glu Asp Vai Gly Asp Ala Ile Lys Pro
195 200 205
Tyr Ile Val His Arg Cys Arg Gin Ser Ile Asn Phe Ser Leu Gln Cys
'210 215 220
Ala Leu Leu Leu Gly Ala Tyr Ser Ser Asp Met His Ile Ser Thr Gln
225 230 235 240
Arg His Ser Arg Gly Thr Lys Leu Arg Lys Leu Ile Leu Ser Asp Glu
245 250 255
Leu Lys Pro Ala His Arg Lys Arg Giu Leu Pro Ser Leu Ser Pro Ala
260 265 270
Pro Asp Thr Gly Leu Ser Pro Ser Lys Arg Thr His Gln Arg Ser Lys
275 280 285
Ser Asp Ala Thr Ala Ser Ile Ser Leu Ser Ser Asn Leu Lys Arg Thr
290 295 300
Ala Ser Asn Pro Lys Val Glu Asn Glu Asp Glu Glu Lau Ser Ser Ser
305 310 315 320
Thr Glu Ser Ile Asp Asn Ser Phe Ser Ser Pro Val Arg Leu Ala Pro
325 330 335
Glu Arg Glu Phe Ile Lys Ser Leu Met Ala Ile Gly Lys Arg Leu Ala
340 345 350
CA 02458872 2004-03-23
-155-
Thr Leu Pro Thr Lys Glu Gln Lys Thr Gin Arg Leu Ile Ser Glu Leu
355 360 365
Ser Leu Leu Asn His Lys Leu Pro Ala Arg Val Trp Leu Pro Thr Ala
370 375 380
Gly Phe Asp His His Val Val Arg Val Pro His Thr Gln Ala Val Val
385 390 395 400
Leu Asn Ser Lys Asp Lys Ala Pro Tyr Leu Ile Tyr Val Glu Val Leu
405 410 415
Glu Cys Glu Asn Phe Asp Thr Thr Ser Val Pro Ala Arg Ile Pro Glu
420 425 430
Asn Arg Ile Arg Ser Thr Arg Ser Val Glu Asn Lsu Pro Glu Cys Gly
435 440 445
Ile Thr His Glu Gin Arg Ala Gly Ser Phe Ser Thr Val Pro Asn Tyr
450 455 460
Asp Asn Asp Asp Glu Ala Trp Ser Val Asp Asp Ile Gly Glu Leu Gln
465 470 475 480
Val Glu Leu Pro Glu Val His Thr Asn Ser Cys Asp Asn Ile.Ser Gin
485 490 495
Phe Ser Val Asp Ser Ile Thr Ser Gin Glu Ser Lys Glu Pro Val Phe
500 505 510
Ile Ala Ala Gly Asp Ile Arg Arg Arg Leu Ser Glu Gin Leu Ala His
515 520 525
Thr Pro Thr Ala Phe Lys Arg Asp Pro Glu Asp Pro Ser Ala Val Ala
530 535 540
Leu Lys Glu Pro Trp Gln Giu Lys Val Arg Arg Ile Arg Glu Gly Ser
545 550 555 560
Pro Tyr Gly His Leu Pro Asn Trp Arg Lsu Leu Ser Val Ile Val Lys
565 570 575
Cys Gly Asp Asp Lau Arg Gln Glu Leu Leu Ala Phe Gln Val Leu Lys
580 585 590
Gln Leu Gln Ser Ile Trp Glu Gln Glu Arg Val Pro LQu Trp Ile Lys
595 600 605
Pro Ile Gln Asp Ser Cys Glu Ile Thr Thr Asp Ser Gly Met Ile Glu
610 615 620
CA 02458872 2004-03-23
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Pro Val Val Asn Ala Val Ser Ile His Gln Val Lys Lys Gln Ser Gln
625 630 635 640
Leu Ser Leu Leu Asp Tyr Phe Leu Gln Glu His Gly Ser Tyr Thr Thr
645 650 655
Glu Ala Phe L8u Ser Ala Gln Arg Asn Phe Val Gin Ser Cys Ala Gly
660 665 670
Tyr Cys Leu Val Cys Tyr Leu Leu Gln Val Lys Asp Arg His Asn Gly
675 680 685
Asn I1e L2u Leu Asp Ala Giu Gly His Ile Ile His Ile Asp Phe Gly
690 695 700
Phe Ile Leu Ser Ser Ser Pro Arg Asn Leu Gly Phe Glu Thr Ser Ala
705 710 715 720
Phe Lys Leu Thr Thr Glu Phe Val Asp Val Met Gly Gly Leu Asp Gly
725 730 735
Asp Met Phe Asn Tyr Tyr Lys Met Leu Met Leu Gln Gly Leu Ile Ala
740 745 750
Ala Arg Lys His Met Asp Lys Val Val Gln Ile=Val Glu Ile Met Gin
755 760 765
Gln Gly Ser Gin Leu Pro Cys Phe His Gly Ser Ser Thr Ile Arg Asn
770 775 780
Leu Lys Glu Arg Phe His Met Ser Met Thr Glu Glu Gln L-au Gln Leu
785 790 795 800
Leu Val Glu Gln Met Val Asp Gly Ser Met Arg Ser Ile Thr Thr Lys
805 810 815
Leu Tyr Asp Gly Phe Gln Tyr Lau Thr Asn Gly Ile Met
820 825
(2) INFORMATION FOR SEQ ID N0:32:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2487 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
CA 02458872 2004-03-23
-157-
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:32:
AT('aAC'ATZCT TG(''aAAIOC'PCG AAG"I'CTOOC.T GTOGCCATGG GAGATACAGT AGZC~"aAOCCT
60
GCCX.X"'..CTi'GA AGCCAACTI'C TGAGCCCAC.T TCTGGOCCAC CAGGGAATAA Ta00GGGPCC 120
CTOC,'TAAGTG TCATCACGGA CXXXXPCGGG GAACTATCAG TGATTGACOC ZGAGGT GCC 180
CAGAAGGCCT OCCAGGAGf3P GPt'0('aAGAAA G'PCAACaCTTT TGCATGGAGG CJ(3TOGCAGrC 240
TCTAGCAGAG GCAIOCCCACT GC'~UGPIGGI'C AAZCGGC~ATG GTGPOGACAG TCAGATCOGP 300
TGCGTAGATG ATOC'.AOCTgC CCAiGATC,AGC' GAGGAGGAAG ATGAC'~TOGG aGCCGCTG'1G 360
OCCiCAQOCA CAGCCAAAGG AOC.AAGAAGA aGGCGOCAIGA ACAACTCAGC TAAAICAGIY.'T 420
TOOCTOCTGA OGCTGIRTGA GICAAAACTG TTTGACATCT CCATOGIX,AT TTCATAfJCTG 480
TATAACl'OCA AOGAOCC'I'GG AGTACAAQCC TACATTOQCA AOOOOCt'IT CTUCITTCOC 540
AACGAOGACG TGGA(.'TTCTA TCTaCCCCAG TTQCZTAACA TGTACATaCA CA2OGATOAG 600
GAOGTOGGIG ATOOCATTAA GCCC.TAlCATA G"POCACOGTT GCCGOC.AGAG CATTAACiTT 660
TOOC'!'CCA~'T GTOOOC.Z'GTP OCTI'C~OC~GCC TATTCTIC'.AG ACATGCA[',AT TTCCACrCAA
720
t7GAC.AC'IY70C C''TOGGAICCAA GC.`PACO('aAAG CTGATCCIY,`P CAGATGAOC.T
AAAIGCC.AGC'P 780
CACAGGAAOA aGC=AOCTGCC CTCCTTGAGC CCGGCCCC'iG ATAC'AGGGC.T G7.'G"PCO(."i'CC
840
AAAAGCAL:1'C AOCAaCGCTC TAHGTI'AC'~AT GCCACZGCCA GCATAAGI'CT CAQCAOCAAC 900
CTtaAAACGAA CAODCAGCAA CCCTAAAG'TG GAGAATGAGG ATGADC'~,AGCT CTOCI'OCAGC 960
ACCC'~AIGAGTA TTGATAATTC ATTCAG'PI'OC CCTGTTOGAC T GCPOCT('aiA GAC'~AGAATTC
1020
ATt;AAO'TCOC TGATOGOC'~P,T CGOCAAGC'GG CTGOCC'ACGC TOCOf'.ACCAA = AGP-GCAGAAA
1080
ACACAGAGGC TGATCZCAGA CZCTOOCPG GTCAACCATA AGC.'TOCCTGC CCGAG't'CIGG 1140
GTOCCCAC'tG CTTOGCTTTGA CCACCACGTG Gi'CCGTGTAC CCCACACACA C~GCPGTI'(:'TC 1200
CI'C'AACTCf'.A AG'GACAAOGC TCCCTACCTG ATTTATGIGG AAG'TOCTIGA ATGTGAAAAC 1260
TTTCA~C'.ACX'A CCACTGTCCC ZGOCCGf'~TC CCCGAGAAQC GAATTCGGAG TACGAGGTCC 1320
GTACAAAAGT 'IGOOCGAAZG TGGPATTACC CATGACCAGC GAQCIGOC,AG GTPC14iGCACr 1380
GI' CC('.AACT AZGAuCAACGA TGATC'~i00CC TGGTCGGTGG A'I'GACATAGG CGAGCTOCAA 1440
CA 02458872 2004-03-23
~
-158-
GZGGAGCTOC CxGAAG'IGCA TACCAAC,AGC TGTGACAACA TC'I'CCCAGTT CTC'IG'I'CGAC 1500
AGCATCACCA GC(',AGGAGAG CAAOGHGCCT GTGTTCATTG CAGC'AGGGGA CAZ'COQCCGG 1560
COOCTTPCGG AACAOC'I'GOC TCATACCCCG ACAGCCPICA AAOGAGACCC AGAAGATCCT 1620
TCIGCAGI'TG CTC1C'.AAAGA GCCCZGGCAG GAGAAAGrAC G CGGATCAG AGAGGGC'IY7C 1680
C(7CTACGGC7C ATC7IC.700CAA TIC~GOOGC,'TC CTC'TCAlGPCA TTGTCAAGIG TOGGGATGAC
1740
CTTOOGCAAG AGCI'TCTGOC G`ITICAGGPG TTGAAGCAAC TGCAG'i'CCAT TIGGGAACAG 1800
GAOCGAQY'GC CC(TIRGGAT CAAG('.C.AATA CAAGATTCTT GPCAAATTAC GAC'iGATAGT 1860
GGCATGATTG AAOCAiGI'GGT CAATGCIG'IG TCCATCCATC AGGTGAAGAA ACAGTCACAG 1920
CTCZC7CT.[GC TCGAZTACTT aC;TAC,AGGAG C'.lAOGGCAGl'T AC'.ACCArCI'GA
GGC'.ATTCC'IC 1980
AGTGCACAGC GCAATTTPGr GCAAAGT'IGT GC'tGGGTACT (~T'i'GGIC`IG CTACC,'ZGC'1G 2040
CAAiG'iCAAQG ACAGACACA,A TOOGAATATC CTTTIGGAiOG CAGAAOGOCA CATCAT'CCAC 2100
ATCGACTPiG GCIRCATCCT CTOCAGCI'CA CCCCGAAATC ZGO CPTTGA, GACGIC'AGCC 2160
TTTAAGCMA CCACAGAGTT TGIUC'1TG7G A'IGGGOGGCC TGGA'i'OOOCaA CATGPIC'=AAC 2220
TACTATAAGA TGGZGATGCT GCAAQGOGRG ATIGOOOC:'iC GGAAACACAT GGACAAGGIG 2280
GTGCAGATCG T GAGATCAT GCAOCAAGGT TGTC'AOCITC CTTGCI'POC.'A TGGC't'CC.AGC 2340
ACCATI'CC'~AA ACCPCAAAGA GAGGTTCCAC ATGAGCATGA CTGAGGADCA GCTOC'AOG`IG 2400
C'TGG'I'GGAIDC AGATG(3TGf`aA ZGOC,AGTATG CGGPCTATCA CC',ACCAAALT C.TATGACGGC
2460
TTCCAG'PAOC TC:AOf'.AAOGG CATCATG 2487
(2) INFORMATION FOR SEQ ID N0:33:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3324 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
CA 02458872 2004-03-23
-159-
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-428B12c1
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 115..2601
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:33:
COGC'aAATTC)C GGGAAGOCXOG GAIOCAAG'TTT TGAAGAAGTC CXTAZCAGAT TACACPI`GGT 60
TGALTACTOC GGAGCAG(X'A CTAAGAGGGA TGAHC AGGGC 1GCGI'GC'~ TTGA ATG 117
Met
1
AGA TTC TTG GP,A GL.`r CGA AGT CTG Gf.'T GTG GOC ATG GGA GAT ACA GTA 165
Arg Phe Leu Glu Ala ArggSer L2u Ala Val Ala Met Gly Asp Thr Val
10 15
GTG GAG OCT GflC COC TTG AAG CCA ACT TCT GAG DOC ACT TLT GGC OCA 213
Val Glu Pro Ala Pro Leu Lys Pro Thr Ser Glu Pro Thr Ser Gly Pro
20 25 30
CCA GGG AAT AAT GGG GflG TCC CTG CTA AC,'P GPC ATC ACG GAG GOG GTC 261
Pro Gly Asn Asn Gly Gly Ser Leu Leu Ser Val Ile Thr Glu Gly Val
35 40 45
GGG GAA CTA TCA GTG ATT GAC CCT GAG GIG GOC CAG AAG GOC TGC CAG 309
Gly Glu Leu Ser Val Ile Asp Pro Glu Val Ala Gln Lys Ala Cys Gln
50 55 60 65
GAG GIG TTG GAG AAA GTC AAG CTT TTG CAT GGA GGC GTG GCA GTC TCT 357
Glu Val Lau Glu Lys Val Lys Leu Leu His Giy Gly Val Ala Val Ser
70 75 80
AGC AGA GflC ACC Cr'A CTG GAG TTG GTC AAT GGG GAT GGT GTG GAC AGT 405
Ser Arg Gly Thr Pro Leu Glu Leu Val Asn Gly Asp Gly Val Asp Ser
85 90 95
GAG AZC CGT TGC C'TA GAT GAT CCA CCT GOC CAG ATC AGG GAG GAG GAA 453
Glu Ile Arg Cys Leu Asp Asp Pro Pro Ala Gln Ile Arg Glu Glu Glu
100 105 110
GAT GAG ATG GGG GCC GCT GTG GCC TCA GGC ACA GOC AAA GGA GCA AGA 501
Asp Glu Met Gly Ala Ala Val Ala Ser Gly Thr Ala Lys Gly Ala Arg
115 120 125
AGA aGG CGG CAG AAC AAC TCA GCT AAA CAG TCT TG+G CTG CTG AGG CIG 549
CA 02458872 2004-03-23
-160-
Arg Arg Arg Gln Asn Asn Ser Ala Lys G1n Ser Trp Leu Lau Arg Lau
130 135 140 145
TTT GAG TCA AAA CTG TTT GAC ATC TCC ATG GOC ATT TCA TAC CTG TAT 597
Phe Glu Ser Lys Leu Phe Asp Ile Ser Met Ala Ile Ser Tyr Leu Tyr
150 155 160
AAC TCC AAG GAG OCT GGA GTA CAA GOC TAC ATT GGC AArC CGG CTC TTC 645
Asn Ser Lys Glu Pro G1y Val Gln Ala Tyr I].e Gly Asn Arg Leu Phe
165 170 175
TGC TTT CGC AAC GAG GAC GIG GAC TTC TAT CTG OOC CAG TTG C1T AAC 693
Cys Phe Arg Asn Glu Asp Val Asp Phe Tyr Leu Pro Gln Iau Leu Asn
180 185 190
ATG TAC ATC CAC ATG GAT GAG GAC GTG OGT GAT GOC ATT AAG UOG TAC 741
Met Tyr Ile His Met Asp Glu Asp Val Gly Asp Ala Ile Lys Pro Tyr
195 200 205
ATA GTG CAC aGT TGC OGC CAG AGiC ART AAC TTT TCC CTC CAG TGr GOC 789
Ile Val His Arg Cys Arg Gln Ser I1e Asn Phe Ser Leu Gin Cys Ala
210 215 220 225
CPG TTG CIT GGG GflC TAT TCT TCA GAC ATG CAC AZTT TCC ACP CAA CGA 837
Lgu Leu Leu Gly Ala Tyr Ser Ser Asp Met His Ile Ser Thr Gin Arg
230 235 240
CAC TCC CGT GflG AOC AAG CTA aGG AAG CPG ATG CIC TCA GAT GAG G'TA 885
His Ser Arg Gly Thr Lys Leu Arg Lys Leu Ile Leu Ser Asp Giu Lau
245 250 255
AAG C7CA GCT CAC AGG AAG AGG GAG CTG OOC TCC TTG AGC OOG GOC CJC'P 933
Lys Pro Ala His Arg Lys Arg Glu Leu Pro Ser Leu Ser Pro Ala Pro
260 265 270
GAT ACA GGG CTG TL'P OOC TCC AAA AGG AGT CAC CAG C7C7C TCT AAC TCA 981
Asp Thr Gly Leu Ser Pro Ser Lys Arg Thr His Gln Arg Ser Lys Ser
275 280 285
GAT GOC ACT GflC AOC ATA AGr CTC AGC AOC AAC CTG AAA aGA ACA GOC 1029
Asp Ala Thr Ala Ser Ile Ser Leu Ser Ser Asn Leu Lys Arg Thr Ala
290 295 300 305
AGC AAC CCT AAA QPG GAG AAT GAG GAT GAG GAG CPC TCC TCC AGC ACC 1077
Ser Asn Pro Lys Val Glu Asn Glu Asp Glu Glu Leu Ser Ser Ser Thr
310 315 320
GAG AG'P ATT GAT AAT TCA TTC AIGT TCC OCT GTT CGA CTG GCT CCT GAG 1125
Glu Ser Ile Asp Asn Ser Phe Ser Ser Pro Val Arg Leu Ala Pro Glu
325 330 335
CA 02458872 2004-03-23
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AGA GAA TTC ATC AAG TCC C'I'G ATG GCG ATC GGC AAG CGG CTG GCC ACG 1173
Arg Glu Phe Ile Lys Ser Leu Met Ala Ile Gly Lys Arg Leu Ala Thr
340 345 350
CTC QOC ACC AAA GAG CAG AAA ACA CAG AGG CIG ATC ZCA GAG C'I'C TCC 1221
Leu Pro Thr Lys Glu Gln Lys Thr Gln Arg Leu Ile Ser Glu Leu Ser
355 360 365
CTG CTC AAC CAT AAG CTC OCT GOC CGA GTC TGG C'IG CCC AG'T GCT GGC 1269
Leu Leu Asn His Lys Leu Pro Ala Arg Val Trp Leu Pro Thr Ala Gly
370 375 380 385
TTT GAC CAC CAC GTG GTC CGT GTA COC CAC ACA CAG GL'T GIT GTC CTC 1317
Phe Asp His His Val Val Arg Val Pro His Thr Gln Ala Val Val Leu
390 395 400
AAC '1'C7C AAAG GAC AAG GO'!' OC7C TAC CTG AZT TAT GTG GAA GTC CTT GAA 1365
Asn Ser Lys Asp Lys Ala Pro Tyr Leu Ile Tyr Val Glu Val Leu Glu
405 410 415
TGT GAA AAC TTT GAC ACC ACC AGT GTC OCT GOC CGG ATC UOC GAG AAC 1413
Cys Glu Asn Phe Asp Thr Thr Ser Val Pro Ala Arg Ile Pro Glu Asn
420 425 430
CGA ATT CGG AGT ACG AGG TOC GTA GAA AAC TTG QOC GAA TGT GGT ATT 1461
Arg Ile Arg Ser Thr Arg Ser Val Glu Asn Leu Pro Glu Cys Gly Ile
435 440 445
ACC CAT GAG CAG CGA GCT GGO AGO TTC AGC ACT GTG OOC AA+C TAT GAC 1509
Thr His Glu Gln Arg Ala Gly Ser Phe Ser Thr Val Pro Asn Tyr Asp
450 455 460 465
AAC GAT GAT GAG GOC TGG TCG GTG GAT GAC ATA GGC GAG CTG CAA GTG 1557
Asn Asp Asp Glu Ala Trp Ser Val Asp Asp Ile Gly Glu Leu Gln Val
470 475 480
GAG CIC QOC GAA GTG CAT ACC AAC AGC TGT GAC AAC ATC TOC CAG TI'C 1605
Glu Leu Pro Glu Vai His Thr Asn Ser Cys Asp Asn Ile Ser Gln Phe
485 490 495
TCT GTG GAC AGC ATC ACC AGC CAG GAG AGC AAG GAG OC.'T GTG 'I'I'C ATT 1653
Ser Val Asp Ser Ile Thr Ser Gln Glu Ser Lys Glu Pro Val Phe Ile
500 505 510
GCA GCA GOG GAC ATC CGC CGG CJGC CTT TL7G GAA CAG CIG GCT CAT ACC 1701
Ala Ala Giy Asp Ile Arg Arg Arg Leu Ser Glu Gln Leu Ala His Thr
515 520 525
OOG ACA GUC TTC AAA CGA GAC C7CA GAA GAT C7CT TC,'T GCA GTT GCT CIC 1749
Pro Thr Ala Phe Lys Arg Asp Pro Glu Asp Pro Ser Ala Val Ala Leu
CA 02458872 2004-03-23
-162-
530 535 540 545
AAA GAG QOC TGG CAG GAG AAA GTA CGG aGG ATC AGA GAG GGC TOC CCC 1797
Lys Glu Pro Trp Gln Glu Lys Val Arg Arg Ile Arg Glu Gly Ser Pro
550 555 560
TAC GGC CAT C'PC OOC AAT TGG aGG CTC C'1'G ZCA GTC ATT GTC AAG TGT 1845
Tyr Gly His Leu Pro Asn Trp Arg Leu Leu Ser Val Ile Val Lys Cys
565 570 575
GGG GAT GAC CZT COG CAA GAG C.'TT GTG Gt7C TTT CAG GTG TTG AAG CAA 1893
Giy Asp Asp Leu Arg Gln Glu Lsu Leu Ala Phe Gln Val Leu Lys Gln
580 585 590
Ct'G CAG TOC ATT TOG GAA CAG GAG CGA GTG DOC CIT TGG ATC AAG OCA 1941
LBu Gln Ser Ile Trp Glu Gln Glu Arg Val Pro Leu Trp Ile Lys Pro
595 600 605
ATA CAA GAT TCT TGT GAA ATT AOG ACT GAT AGT GGC ATG ATT GAA OCA 1989
Ile Gln Asp Ser Cys Glu Ile Thr Thr Asp Ser Gly Met Ile Glu Pro
610 615 620 625
G=PG GTC AAT GCT GTG TCC ATC CAT CAG GTG AAG AAA CAG ZCA CAG C'I'C 2037
Val Val Asn Ala Val Ser Ile His Gln Val Lys Lys Gln Ser Gln Lau
630 635 640
TOC TTG CTC GAT TAC TTC CTA CAG GAG CAC GGC AGT TAC ACC ACT GAG 2085
Ser Leu Leu Asp Tyr Phe Leu Gln Glu His Gly Ser Tyr Thr Thr Glu
645 650 655
GCA TTC C'IPC AGT GCA CAG COC AAT TZT GTG CAA AGT ZGT GCT GGG TAC 2133
Ala Phe Leu Ser Ala Gln Arg Asn Phe Val Gln Ser Cys Ala Gly Tyr
660 665 670
TGC TTG GTC TGC TP,C CTG CIG CAA GTC AAG GAC AGA CAC AAT GGG AAT 2181
Cys Leu Val Cys Tyr Leu Leu Gln Val Lys Asp Arg His Asn Gly Asn
675 680 685
ATC CIT T'I'G GAC GC',A GAA GGC CAC ATC ATC CAC ATC GAC TTT GGC TTC 2229 '
Ile Leu Leu Asp Ala Glu Gly His Ile Ile His Ile Asp Phe Gly Phe
690 695 700 705
ATC CTC TOC AGC TCA COC CGA AAT CTG GflC TTT GAG ACG TCA GOC TTT 2277
Ile Leu Ser Ser Ser Pro Arg Asn Leu Gly Phe Glu Thr Ser Ala Phe
710 715 720
AAG GTG ACC ACA GAG TTT GTG GAT GTG ATG GGC GGC CIG GAT GOC GAC 2325
Lys Leu Thr n-ir Glu Phe Val Asp Val Met Gly Gly Leu Asp Gly Asp
725 730 735
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ATG T'i'C AAC TAC TAT AAG ATG CTG ATG CTG CAA GGG CTG ATT GCC GCT 2373
Met Phe Asn Tyr Tyr Lys Met LSu Met Leu Gln G1y.Leu Ile Ala Ala
740 745 750
OGG AAA CAC ATG GAC AAG GTG GTG CAG ATC GTG GAG ATC ATG CAG CAA 2421
Arg Lys His Met Asp Lys Val Val Gin Ile Val Glu Ile Met Gln Gln
755 760 765
GGT TCT CAG CIT CCT TGC TTC CAT GGC TCC AGC A(:C ATT CGA AAC GTC 2469
Gly Ser Gln Leu Pro Cys Phe His Gly Ser Ser Thr Ile Arg Asn Leu
770 775 780 785
AAA GAG AGG TIC CAC ATG AGC ATG ACT GAG GAG CAG CI'G CAG CTG CPG 2517
Lys Glu Arg Phe His Met Ser Met Thr Glu Glu Gln Leu Gln Leu Leu
790 795 800
GTG GAG CAG ATG CTIG GAT GGC AGT ATG COG TG`T ATC ACC ACC AAA CTC 2565
Val Glu Gln Met Val Asp Gly Ser Met Arg Ser Ile Ztr Thr Lys L8u
805 810 815
TAT GAC GGC TTC CAG TAC CTC ACC AA+C GGC ATC ATG TGA CACGC'lYX.'TC 2614
Tyr Asp Gly Phe GlnTyr Leu Thr Asn Gly Ile Met *
820 825 830
AGCCCAGGAG TGGTG~GOOOG TCCAGOOC'AC CX.*tC70GTAGA GGGOOC1'TGT CZGAGAAA[JC 2674
CCAAAQCAGG AAA+COOCA0C TACCC.APuCCA TOCAOCCAAG GGAAATOGfi-A GGCAA['aAAAIC 2734
ACGAAGGATC AZGTGGTAAC TGOGAG,AGCT TOCIC'AOGf G TGGGAiGAGCC AGCTQI'OOGG 2794
TCCAGACT'1'G TTGGGGCITC CCTGCXC(,'TC CIGGPCTG'I'G TCAGTATTAC CACCAGACTG 2854
ACZCCAGGAiC TCAf.'TGCfx'F CCAGAAAACA GAIOGTGACAA ATGTGAOOC',~A CACTOOGGCC 2914
TTPCTTCTOC TTGTAGQOf't' CTC:'1'CAGAIOG TTCTTPOCAC ApGOC'ATaCT CTTATPCCGT 2974
ZC'IC~OC~f,iCOC AGGAAG!'OGG GAA('+,dGPAGG TPC'!'COG'PAC TTAGGACTTG ATOCZG'IWT
3034
TGCCAC=IGGC CAZGC'IGCPG COCAGC'II'CTA COCCTCOCAG GGACCTAOOC CTCIXCAGf'GA 3094
CCGACOCC'I'G GCCC'.AAGC'I'C COCTTC'CI'GG CGGGCOC,'Z'GC G'7.'GOGCCCi'G
CACTTGCTC'xA 3154
GGT'POCOC'AT CA'IC'OGCAAG GCAAGGGAAT TCCCAC,AGOC CTC7CAG'I"GPA CRGAi00GTAC
3214
TGOOCTAGOC ATGTGGAATT CCCPACCC'1G AC'POCITCCC CAAACCC'=AGC' GAAA~'aAGGT 3274
C'I''.AATPZTP TATZTITAAT TZTTGI'TPGA, AATAAAGIC7C TTAGITAGCC 3324
(2) INFORMATION FOR SEQ ID N0:34:
CA 02458872 2004-03-23
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(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 810 amino acids
(B) TYPE: amino acid
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:
Met Pro Met Asp Leu Ile Leu Val Val Trp Phe Cys Val Cys Thr Ala
1 5 10 15
Arg Thr Val Val Gly Phe Gly Met Asp Pro Asp Leu Gln Met Asp Ile
20 25 30
Val Thr Glu Leu Asp leu Val Asn Thr Thr leu Gly Val Ala Gln Val
35 40 45
Ser Gly Met His Asn Ala Ser Lys Ala Phe leu Phe Gln Asp I].e Glu
50 55 60
Arg Glu Ile His Ala Ala Pro His Val Ser Glu Lys Leu Ile Gln leu
65 70 75 80
Phe Gin Asn Lys Ser Glu Phe Thr Ile Leu Ala Thr Val Gln Gln Lys
85 90 95
Pro Ser Thr Ser Gly Val Ile Leu Ser Ile Arg Glu Leu Glu His Ser
100 105 110
Tyr Phe Glu Leu Glu Ser Ser Gly Leu Arg Asp Glu Ile Arg Tyr His
115 120 125
Tyr Ile His Asn Gly Lys Pro Arg Thr Glu Ala Leu Pro Tyr Arg Met
130 135 140
Ala Asp Gly Gln Trp His Lys Val Ala Leu Ser Val Ser Ala Ser His
145 150 155 160
Leu Leu Leu His Val Asp Cys Asn Arg Ile Tyr Glu Arg Val Ile Asp
165 170 175
Pro Pro Asp Thr Asn Leu Pro Pro Gly Ile Asn Leu Trp Leu Gly Gln
180 185 190
Arg Asn Gln Lys His Gly Leu Phe Lys Gly Ile Ile Gln Asp Gly Lys
195 200 205
Ile Ile Phe Met Pro Asn Gly Tyr Ile Thr Gln Cys Pro Asn Lgu Asn
CA 02458872 2004-03-23
-165-
210 215 220
His Thr Cys Pro Thr Cys Ser Asp Phe Leu Ser Leu Val Gln Gly Ile
225 230 235 240
Met Asp Leu Gin Glu Leu Leu Ala Lys Met Thr Ala Lys Leu Asn Tyr
245 250 255
Ala Glu Thr Arg Leu Ser Gin Leu Glu Asn Cys His Cys Glu Lys Thr
260 265 270
Cys Gln Val Ser Gly Leu Leu Tyr Arg Asp Gln Asp Ser Trp Val Asp
275 280 285
Gly Asp His Cys Arg Asn Cys Thr Cys Lys Ser Gly Ala Val Glu Cys
290 295 300
Arg Arg Met Ser Cys Pro Pro Leu Asn Cys Ser Pro Asp Ser Leu Pro
305 310 315 320
Val His Ile Ala Gly Gin Cys Cys Lys Val Cys Arg Pro Lys Cys Ile
325 330 335
Tyr Gly Gly Lys Val Leu Ala Glu Gly Gln Arg Ile Leu Thr Lys Ser
340 345 350
Cys Arg Glu Cys Arg Gly Gly Val Leu Val Lys Ile Thr Glu Met Cys
355 360 365
Pro Pro Leu Asn Cys Ser Glu Lys Asp His Ile Leu Pro Glu Asn Gln
370 375 380
Cys Cys Arg Val Cys Arg Gly His Asn Phe Cys Ala Glu Gly Pro Lys
385 390 395 400
Cys Gly Glu Asn Ser Glu Cys Lys Asn Trp Asn Thr Lys Ala Thr Cys
405 410 415
Glu Cys Lys Ser Gly Tyr Ile Ser Val Gln Gly Asp Ser Ala Tyr Cys
420 425 430
Glu Asp Ile Asp Glu Cys Ala Ala Lys Met His Tyr Cys His Ala Asn
435 440 445
Ttir Val Cys Val. Asn Leu Pro Gly LSu Tyr Arg Cys Asp Cys Val Pro
450 455 460
Gly Tyr Ile Arg Val Asp Asp Phe Ser Cys Thr Glu His Asp Glu Cys
465 470 475 480
CA 02458872 2004-03-23
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Gly Ser Gly Gln His Asn Cys Asp Glu Asn Ala Ile Cys Thr Asn Thr
485 490 495
Val Gln Gly His Ser Cys Thr Cys Lys Pro Gly Tyr Val Gly Asn Gly
500 505 510
Thr Ile Cys Arg Ala Phe Cys Glu Glu Gly Cys Arg Tyr Gly Gly Thr
515 520 525
Cys Val Ala Pro Asn Lys Cys Val Cys Pro Ser Gly Phe Thr Gly Ser
530 535 540
His Cys Glu Lys Asp Ile Asp Glu Cys Ser Glu Gly Ile Ile Glu GYs
545 550 555 560
His Asn H;L.s Ser Arg Cys Val Asn Leu Pro Gly Trp Tyr His Cys Glu
565 570 575
Cys Arg Ser Gly Phe His Asp Asp Gly Thr Tyr Ser Leu Ser Gly Glu
580 585 590
Seac Cys Ile Asp Ile Asp Glu Cys Ala Leu Arg Thr His Thr Cys Trp
595 600 605
Asn Asp Ser Ala Cys Ile Asn Leu Ala Gly Gly Phe Asp Cys Leu Cys
610 615 620
Pro Ser Gly Pro Ser Cys Ser Gly Asp Cys Pro His Glu Gly Gly Leu
625 630 635 640
Lys His Asn Gly Gin Val Trp Thr Lau Lys Giu Asp Arg Cys Ser Val
645 650 655
Cys Ser Cys Lys Asp Gly Lys Ile Phe Cys Arg Arg Thr Ala Cys Asp
660 665 670
Cys Gin Asn Pro Ser Ala Asp Leu Phe Cys Cys Pro Glu Cys Asp Thr
675 680 685
Arg Val Thr Ser Gln Cys Leu Asp Gln Asn Gly His Lys Leu Tyr Arg
690 695 700
Ser Gly Asp Asn Trp Thr His Ser Cys Gin Gin Cys Arg Cys Leu Glu
705 710 715 720
Gly Glu Val Asp Cys Trp Pro Leu Thr Cys Pro Asn Leu Ser Cys Glu
725 730 735
Tyr Thr Ala Ile Leu Glu Gly Glu Cys Cys Pro Arg Cys Val Ser Asp
740 745 750
CA 02458872 2004-03-23
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Pro Cys Leu Ala Asp Asn Ile Thr Tyr Asp Ile Arg Lys Thr Cys Leu
755 760 765
Asp Ser Tyr Gly Val Ser Arg Leu Ser Gly Ser Val Trp Thr Met Ala
770 775 780
Gly Ser Pro Cys Thr Thr Cys Lys Cys Lys Asn Gly Arg Val Cys Cys
785 790 795 800
Ser Val Asp Phe Glu Cys Leu Gln Asn Asn
805 810
(2) INFORMATION FOR SEQ ID N0:35:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2430 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:
ATGCCXGATGG ATTTGATTZT AGI'I'Gi'Gi'GG GCAC'i'GCCAC GACAGiGGi'G 60
GGL`ITTGGGA TGGACCCT('aA CCTTC'AGATG GATATCGI'CA CCGAGC,'TTGA CCTTGTGAAC 120
ACCACCCT'IG GAIG'I'I'GC'i'CA GGTGIC;TGGA AZGCACAATG CCAGCAAAGC ATTZTPATTT 180
CAAGACATAG AAAGAGAGAT CCATGCAGCT CCI"CATGZGA GTGAGAAATT AATTC',AGC'IG 240
T'I'OCAGAACA AGAGI'GAATT CACCATPI'ZG GCCAC'i'GTAC AGC.AGAAOOC ATCCAC'ITCA 300
GGAG'Tf'~ATAC TGI'OCATTCG AGAACTOGAG CACAGCTATT TI'C'aAAC."iC'GGGA GAGCAGTGGC
360
CTGAGGGATG AGAT'POGGTA TCACTACATA CACAATGGC'~,A AGCCAAGGAC AGAGGCAC'IT 420
CCZTACCOCA TGGCAGATGG ACAAZGGCAC AAGGrPGCAC TGZCAGI'TAG CGCC'IY,TCAT 480
CTCCI'GC'I'CC ATGPCGACTG TAACAGGATT TATGAGO('TG TGATAGACCC TCCAGATACC 540
AAlOC.`TTCOOC CAGGAATCAA TPPATG+GC,'TT GGOCAGOGCA ACGAAAAGCA TGG(,'TTATTC 600
AAAGGGATCA TCCAAGATGG GAAGAZCATC TTTAZGOOC'~A ATGGATATAT AACACAGTGT 660
CC',AAATC'I'AA ATCACACTZG CCCAACC.'TGC AG=PC'~A'ITI'CP TAAGCC'POGr
GCAAGC'aAATA 720
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ATG{"~ATTTAC AAGAGCPPTT GGCCAAGATG ACPGCAAAAC TAAATTAZGC AGAGACAAGA 780
CTTAGPCAAT TGGAAAACTG TCATTGIPC',AG AAGACrl= AAGTGAGTGG ACTGCPCTAT 840
CGAGATCAAG ACPCITGGGT AGATGGIC3AC CATTOCAiOGA AC'I'OCACTTG CAP-AAGTOGT 900
GCW'TGGAAT GCOGAAGGAT GTOCJ[GZC7CC CCTC'iCAATT GCTCCCCAGA CI'CCCTCO('.A 960
GTACACATTG C'!.'GGOCAGIG CTGTAAOGI'C TGOCC'~~OCAA AATGTATCTA TGGAGGAAAA 1020
GTT'CP'IGC.AG AAQGOC'.AGCG GATITTAACC AAGAGC'I= GGC'aAAZGCCG AGGTGGACTT 1080
TTA+GTAAAAA TTACAC'aAAAT GZ~iC7CTCCP ZTC`aAACTG(,'T CAGAAAAG(''P- TCAICATTCTT
1140
CCTGAGAATC AGI'GC.''1'GCOG TQiCTG'PAGA GQI'CATAAL'T ZTTGTOCA~'*A AGGACCPAAA
1200
TGTGGI''aAAA ACTCAGAGTG CAAAAACTGG AATACAAAAG CTACTIC7I'GA GTGCAAGAGr 1260
GG'i'TACATCr CPGIC7CAGGG P,GACi'CPGCC TACrGT('~AiAG ATATTGATGA G"I'G'1GCAGCT
1320
AAGAZGCATT AGTCa'tC'ATGC CAATACIGIG TG7G'1CAAOC TTOCPGOGTT ATATCGCi'GP 1380
GACTGTG7.CC CAOf'~ATACAT TCGIrGPGGAT GACt"I'GTCTP GTACAC'~AACA OGAT('aAATGI'
1440
G f',AGCGCC AGCACAAC1G TGATGAGAAT GtJCATCIGC'A CCAALA= CC.AGCGACAC 1500
AGC,"I'GC'AIOCT GCAAAOpGGG CTACG-rGGGG AACGGGAOCA TCiGCAGAGC TTTCTGTGAA 1560
GA GGC7,,GCA GATACGf"!'GG AACGZCTGIG GCZC70CAACA AATGT(~'ZK,'1'G TOC'.ATCTOGA
1620
TTCA+CAGC'AA GCf'.ACI'GOC''~A GAAAIGA.TATT GATGAATG'PP CAGAGCGAAT CATTGAG=
1680
C'ACAACC'ATT CCCGCPGO(~`T TAACX.TGCCA GGGTGQPAOC ACTGrGAGrG CA~',AAGOGCT 1740
T'POC`,ATGAIC7G ATGGGALCPA TZCAcR.'G']CC GGGC'AG'rOCT GTATTGAcAT TGATGAATGT
1800
GOCT`PAAGAA CI't`.AiCACCIG TTGC'aAAOGP,T TCTGCCTG{'A TC AA<7CrGGC AGGGGGTPZT
1860
, GOCCCP(,`TGG GCO(.,~I'OCPGC TCIGC;PGACT GPCCTCATGA AGGGGGGCI'G 1920
AAGiCACAATG CCC'AGGIGPG GACCTI'GAAA GAAGACAGGT GTTCTGTCTG CTOCTOCAAG 1980
GATGOC.AAGA TATi'CZGOCG ACGGACAiQCT TGTGATTGOC AGAATCC.AAG TGCTGACC'PA 2040
TTC~~C CAGAAZG'IC'~A CAOCAGAC"IC ACAAGP('.AAT GT1TAGoACCA AAATGGPCAC 2100
AADCTC'!'ATC GAAGi'GGAGA CAATTGGACC CATAGCIGI'C AIGCACTGI'CG GPGTCPGGAA 2160
GGAGAGGI'AG AT'IGCTGGCC ACTCAGTI'GC OCAACI''C'~A GC'1'GTGAGPA TACAGCTATC 2220
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TTAGAAGGGG AATGTTGI'UC CCGCI'GTGTC AG'I`GACCCCT GCCTAGCI'C,A TAACA'ICACC 2280
TATGACATCA GAAAAAC'I'I'G CCTGGACAGC TATGGTGITT C'ACGGL'PPAG TGGC4CAG'iG 2340
TGGACGATGG CTGCATC'IC7C CTGCAC'.,AACC TG'TAAATGCA AGAATGGGAAG AGICZ'GTTGT 2400
TCTGTGGATT TTt'aAG']['GTC'r TC'AAAATAAT 2430
(2) INFORMATION FOR SEQ ID N0:36:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2977 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-073E07
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 103..2532
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:
TAGCAAGTTP GGCGGC'!'OCA AGCCAGGCGC GCC'!'CAGGAT CCAGGCTCAT TTGC'I'TOCAC 60
CTAGCTII'CGG TGCCCCCTGC TAGGCGGGGA CCC.'lC7GAGAG CG ATG CCG ATG GAT 114
Met Pro Met Asp
1
T'IG ATT TTA GTT GTG TGG TTC TGT GTG TGC ACT GOC AGG ACA GTG GTG 162
Leu Ile Leu Val Val Trp Phe Cys Val Cys Thr Ala Arg Thr Val Val
10 15 20
GGC TTT GGG ATG GAC CCT GAC CTT CAG ATG GAT ATC GTC ACC GAG CTT 210
Gly Pte Gly Met Asp Pro Asp Leu Gln Met Asp Ile Val Thr Glu Leu
25 30 35
GAC CTT GTG AAC ACC ACC C'IT GGA GT'T GCT CAG GTG TCT GGA ATG CAC 258
Asp Leu Val Asn Thr Thr Leu Gly Val Ala Gln Val Ser Gly Met His
CA 02458872 2004-03-23
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40 45 50
MT GOC AGC AAA GCA TTT TTA TTT CAA GAC ATA GAA AGA GAG ATC CAT 306
Asn Ala Ser Lys Ala Phe Leu Phe Gln Asp Ile Glu Arg Glu Ile His
55 60 65
GCA GC,'T OCT CAT GTG AGT GAG AAA TTA ATT CAG C'IG TTC CAG AAC AAG 354
Ala Ala Pro His Val Ser Glu Lys Leu Ile Gln Leu Phe Gln Asn Lys
70 75 80
AGT GAA TTC ACC ATT TTG G{jC ACT GTA CAG CAG AAG UCA TOC ACT TCA 402
Ser Glu Phe Thr Ile Leu Ala Thr Val Gin Gin Lys Pro Ser Thr Ser
85 90 95 100
GGA GTG ATA CTG TCC ATT OGA GAA CTG GAG CAC AGC TAT TTT GAA CTG 450
Gly Vai Ile Lau Ser Ile Arg Glu Leu Glu His Ser Tyr Phe Glu Leu
105 110 115
GAG AGC AGT GOC CIG AGG GAT GAG ATT OOG TAT CAC TAC ATA CAC AAT 498
Glu Ser Ser Gly Leu Arg Asp Glu Ile Arg Tyr His Tyr Ile His Asn
120 125 130
GOG AAG OCA AGG ACA GAG GCA CIT (7CT TAC aGC ATG GCA GAT GGA CAA 546
Gly Lys*Pro Arg Thr Glu Ala Leu Pro Tyr Arg Met Ala Asp Gly Gln
135 140 145
TGG CAC AAG GTT GCA C't'G TCA GTT AGC GOC TOT CAT CTC CrG CTC CAT 594
Trp His Lys Val Ala Leu Ser Val Ser Ala Ser His Leu Leu Leu His
150 155 160
GI'C GAC TGT AAC AaG ATP TAT GAG OGT GTG ATA GAC CGT CCA GAT AOC 642
Val Asp Cys Asn Arg Ile Tyr Glu Arg Val Ile Asp Pro Pro Asp Zhr
165 170 175 180
AAC CIT OOC CCA GGA ATC AAT TTA TGG CIT GGC CAG CGC AAC CAA AAG 690
Asn Lsu Pro Pro Gly Ile Asn Leu Trp L8u Gly Gln Arg Asn Gln Lys
185 190 195
CAT GGC TTA TIC AAA GGG ATC ATC CAA GAT GGG AAG ATC ATC TTT ATG 738
His Gly Leu Phe Lys Gly Ile Ile Gln Asp Gly Lys Ile Ile Phe Met
200 205 210
OOG AAT GGA TAT ATA ACA CAG TGT CCA AAT GTA AAT CAC ACT TOC OCA 786
Pro Asn Gly Tyr Ile Thr Gln Gys Pro Asn Leu Asn His Thr Cys Pro
215 220 225
ACC TGO AGT GAT TTC TTA AGC C'I`G GTG CAA GGA ATA ATG GAT TTA CAA 834
Thr Cys Ser Asp Phe Leu Ser Leu Val Gin Gly Ile Met Asp Leu Gin
230 235 240
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GAG CTT TTG GCC AAG ATG ACT GCA AAA CTA MT TAT GCA GAG ACA AGA 882
Glu Leu Leu Ala Lys Met Thr Ala Lys Leu Asn Tyr Ala Glu Thr Arg
245 250 255 260
C,''iT AGP CAA T'iG GAA AAC TGT CAT TGT GAG AAG ACT TGT CAA GTG AGT 930
Leu Ser Gln Lsu Glu Asn Cys His Cys Glu Lys Thr Cys Gln Val Ser
265 270 275
GGA CTG CTC TAT OGA GAT CAA GAC TCT TGG GTA GAT GGT GAIC CAT ZGC 978
Gly Leu LSu Tyr Arg Asp Gln Asp Ser 'i'zp Val Asp Gly Asp His qis
280 285 290
AGG AlAG = P,CT TCaC AAA AGT GGT GOC GTG GAA RGC OGA AGG ATG TCC 1026
Arg Asn Cys Thr Cys Lys Ser Gly Ala Val Glu Cys Arg Arg Met Ser
295 300 305
TGT CCC aCT CTC AAT TOC TOC OCA GAC TOC CTC OCA GTA CAC ATT GCT 1074
Cys Pro Pro Leu Asn Cys Ser Pro Asp Ser L8u Pro Val His Ile Ala
310 315 320
GflC CAG TOC TGT AAG GTC TGC OGA CCA AAA TGT ATC TAT GGA GGA AAA 1122
Gly Gin Cys Cys Lys Val Cys Arg Pro Lys Cys Ile Tyr Gly Gly Lys
325 330 335 340
GTT CTT GCA GAA GOC CAG CGG ATT TTA AQC AAIG AGG TGT CGG GAA TGC 1170
Val Leu Ala Glu Gly Gln Arg Ile Lsu Thr Lys Ser Cys Arg Glu Cys
345 350 355
CXaA GGT GGA GTT TPA GTA AAA ATT ACA GAA ATG TGT aCT aCT TTG AAC 1218
Arg Gly Gly Val Leu Val Lys Ile Thr Glu Met Cys Pro Pro Leu Asn
360 365 370
TGC TCA GAA AAG GAT CAC ATT CTT C7GT GAG AAT CAG TGC TGC OGT GTC 1266
Cys Ser Glu Lys Asp His Ile Leu Pro Glu Asn Gln Cys Cys Arg Val
375 380 385
TG'T AGA GGT CAT AAC TTT TGT GCA GAA GGA aCT AAA TGT GGT GAA AAC 1314
Cys Arg Gly His Asn Phe Cys Ala Glu Gly Pro Lys GYs Gly Glu Asn
390 395 400
TCA GAG TGC AAA AAC T{zG AAT ACA AAA GC.T ACT ZGP GAG TGC AAG AG'T 1362
Ser Glu Cys Lys Asn Trp Asn Thr Lys Ala Thr Cys Glu Cys Lys Ser
405 410 415 420
GGT TAC ATC TCT GTC CAG GGA GAC 'ICT GflC TAC TGT GAA GAT ATT GAT 1410
Gly Tyr Ile Ser Val Gln Gly Asp Ser Ala Tyr Cys Glu Asp Ile Asp
425 430 435
GAG TGT GCA GCT AAG ATG CAT TAC TGT CAT GOC AAT ACT G'TG TGT GTC 1458
Glu Cys Ala Ala Lys Met His Tyr Cys His Ala Asn Thr Val Cys Val
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440 445 450
AAC CTT CCT GGG TTA TAT CGC TGT GAC TGT GTC CCA GGA TAC AZT aG'I' 1506
Asn Leu Pro Gly Leu Tyr Arg Cys Asp Cys Val Pro Gly Tyr Ile Arg
455 460 465
GTG GAT GAC TTC TCT TGT ACA GAA CAC GAT GAA TGT GGC AGC GGC CAG 1554
Val Asp Asp Phe Ser Cys Thr Glu His Asp Glu Cys Gly Ser Gly Gln
470 475 480
CAC AAC TGT GAT GAG AAT GOC ATC TGC ACC AAC lAGT GTC CAG GGA CAC 1602
His Asn Cys Asp Glu Asn Ala Ile Cys Thr Asn Thr Val Gln Gly His
485 490 495 500
AGC TGC ACC ZM AAA a0G GGC TAC GTG GGG AAC GGG ACC ATC TGC AGA 1650
Ser Cys Thr Cys Lys Pro Gly Tyr Val Gly Asn Gly Thr Ile Cys Arg
505 510 515
GCT TZ'C TGT GAA GAG GOC TOC AGA TAC GGT GGA ACG TGT GTG GCT QOC 1698
Ala Phe Cys Glu Glu Gly Cys Arg Tyr Gly Gly Thr Cys Val Ala Pro
520 525 530
AAC AAA TGT GTC TGT CX'A TC,T GGA TTC ACA GGA AGC CAC TGC GAG AAA 1746
Asn Lys Cys Val Cys Pro Ser Gly Phe Thr Gly Ser His Cys Glu Lys
535 540 545
GAT ATT GAT GAA TGT TCA GAG GGA ATC AZT GAG TGC CAC AAC CAT TOC 1794
Asp Ile Asp Glu Cys Ser Glu Gly Ile Ile Glu Cys His Asn His Ser
550 555 560
GGC TGC GTT AAC CIG aCA GGG TGG TAC CAC TGr GAG TGC AGA AGC GGT 1842
Arg Cys Val Asn Leu Pro Gly Trp Tyr His Cys Glu Cys Arg Ser Gly
565 570 575 580
TPC CAT GAC GAT GtflG ACC TAT TCA CTG TOC GGG GAG TOC TGT ATT GAC 1890
Phe His Asp Asp Gly Thi- Tyr Ser Leu Ser Gly Glu Ser Cys Ile Asp
585 590 595
ATT GAT GAA TGT GCC TTA AGA ACT CAC ACC TGT TGG AAC GAT TCT GOC 1938
Ile Asp Glu Cys Ala Leu Arg Thr His Thr G`ys Tip Asn Asp Ser Ala
600 605 610
TGC ATC AAC CTG GOA GGG GGT TTT GAC TGT C'I'C TGC CCC TC,T GGG CCC 1986
Cys Ile Asn Leu Ala Gly Gly Phe Asp Cys Lau Cys Pro Ser Gly Pro
615 620 625
TC7C TGC TCT GGT GAC TGT CCI' CAT GAA GOG GOG CI'G AAG CAC AAT GGC 2034
Ser Cys Ser Gly Asp Cys Pro His Glu Gly Gly Leu Lys His Asn Gly
630 635 640
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CAG GTG TGG A.OC TTG AAA GJ4A GAC AGG TGT TCT GTC TGC Tt7C TGC AAG 2082
Gln Val Trp Thr Leu Lys Glu Asp Arg Cys Ser Val Cys Ser Cys Lys
645 650 655 660
GAT GGC AAG ATA TTC TGC OGA taGG ACA GCT TGT GAT TaC CAG AAT aCA 2130
Asp Gly Lys Ile Phe Cys Arg Arg Thr Ala Cys Asp Cys Gln Asn Pro
665 670 675
AGT CaCT GAC CTA TTC TGT TGC CCA GAA TGT GAC ACC AGA GTC ACA AQr 2178
Ser Ala Asp Leu Phe Cys Cys Pro Glu Cys Asp Thr Arg Val Thr Ser
680 685 690
CAA TGT TTA GAC CAA AAT GGT CAC AAG CiG TAT t7GA AGT GGA GAC AAT 2226
Gin Cys Leu Asp Gln Asn Gly His Lys Leu Tyr Arg Ser Gly Asp Asn
695 700 705
TGC ACC CAT AGC TGT CAG CAG TGT CGG TGP CI'G GAA GGA GAG GTA GAT 2274
Tip Thr His Ser Cys Gin Gin Cys Arg Cys Leu Glu Gly Glu Val Asp
710 715 720
TOC TGG CCA CTC ACT TGC aCC AAC TTG AOC TGT GAG TAT ACA GGT ATG 2322
Cys Trp Pro Ieu Thr Cys Pro Asn. Leu Ser Cys Glu Tyr Thr Ala Ile
725 730 735 740
TTA GAA GGG GAA TGT ZGT CCC CGC TGT GTC AGT GAC CCC TaC CTA GCT 2370
Leu Glu Gly Giu Cys Cys Pro Arg Cys Val Ser Asp Pro Cys Leu Ala
745 750 755
GAT AAC ATG ACC TAT GAC ATG AGA AAA AGT TGC CTG GAC AGC TAT GGT 2418
Asp Asn Ile Thr Tyr Asp Ile Arg Lys Thr Cys Leu Asp Ser Tyr Gly
760 765 770
GTT TCA OGG CTT AGT GGC TCA GTG TGG AaG ATG GGT GGA TGT CCC TGC 2466
Val Ser Arg Leu Ser Gly Ser Val Trp Thr Met Ala Gly Ser Pro Cys
775 780 785
ACA ACc TG'P AAA TGC AAG AAT GGA AGA GTC TGT TGT TC'I' GTG GAT TTT 2514
Thr Thr Cys Lys Cys Lys Asn Gly Arg Val Cys Cys Ser Val Asp Phe
790 795 800
GAG TGT CTT CAA AAT AAT ZGAA~'~TAT'PT ACAGTOGAlCT CAACXGCAGAA 2562
Glu Cys Leu Gin Asn Asn
805 810
GAAZGGACC,A AATGACCP-TG CAACGPGATT AAOGATAGGA ATOOGPAGTT ZGGITFITTT 2622
GTTi'GI'T"!.*PG TTTI'I`ITAAC CACAGATAAT TGCCAAAGTT 'ICCACCTGAG GA+OOGTGI'TT
2682
aGGAGG'I'TGC CT'1'TTGC'~A+OC TACCA,CPTTG C,'TC'ATTCP'IG CTAACCTAGr CTAGGPGAiCC
2742
CA 02458872 2004-03-23
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TAC;AGTOCCG TGC'ATTTAAG TCAATOGTI'G TTAAAAGAAG ZTTOOGG'TGP TGTAAATCAT 2802
GTT'r'(ACTTA TC'AIGATCATT TGCAAP-TACA TTTAAATGAT CTCATOGTAA ATOGTI'GATG 2862
TAT'I'Z*I't'TGG GTTTATTTIG TGrAG'TAACC ATAATAGAGA GAGACTCAGC TC{.`1TITATT 2922
TATT'I'TG'TTG ATTTATGGAT CAAATTCTAA AATAAAQPIG CCTGr'rG'IGA CT~PT 2977
(2) INFORMATION FOR SEQ ID NO:37:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 816 amino acids
(B) TYPE: amino acid
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:
Met Glu Ser Arg Val Leu Leu Arg Thr Phe Cys Leu Ile Phe Gly Leu
1 5 10 15
Gly Ala Val Trp Gly Leu Gly Va1 Asp Pro Ser Leu Gin Ile Asp Val
20 25 30
Leu Thr Glu Leu Glu Leu Gly Glu Ser Thr Thr Gly Val Arg Gln Val
35 40 45
Pro Gly Leu His Asn Gly Thr Lys Ala Phe Lgu Phe Gln Asp Thr Pro
50 55 60
Arg Ser ile Lys Ala Ser Thr Ala Thr Ala Glu Gln Phe Phe Gln Lys
65 70 75 80
Leu Arg Asn Lys His Glu Phe Thr Ile Leu Val Thr Lau Lys Gln Thr
85 90 95
His Leu Asn Ser Gly Val Ile Leu Ser Ile His His Leu Asp His Arg
100 105 110
Tyr Leu Glu Leu Glu Ser Ser Gly His Arg Asn Glu Val Arg Leu His
115 120 125
Tyr Arg Ser Gly Ser His Arg Pro His Thr Glu Val Phe Pro Tyr Ile
130 135 140
Lsu Ala Asp Asp Lys Trp His Lys Leu Ser Lsu Ala Ile Ser Ala Ser
CA 02458872 2004-03-23
-175-
145 150 155 160
His Leu Ile Leu His I1e Asp Cys Asn Lys Ile Tyr Glu Arg Val Val
165 170 175
Glu Lys Pro Ser Thr Asp Leu Pro Lau Gly Thr Thr Phe Trp Leu Gly
180 185 190
Gln Arg Asn Asn Ala His Gly Tyr Phe Lys Gly Ile Met Gln Asp Val
195 200 205
Gln Leu Leu Val Met Pro Gln Giy Phe Ile Ala Gln Cys Pro Asp Leu
210 215 220
Asn Arg Z'hr Cys Pro Thr Cys Asn Asp Phe His Gly Leu Val Gln Lys
225 230 235 240
Ile Met Glu Leu Gin Asp Ile Leu Ala Lys Thr Ser Ala Lys Leu Ser
245 250 255
Axg Ala Glu Gln Arg Met Asn Arg Leu Asp Gln Cys Tyr Cys Glu Arg
260 265 270
Thr Cys Thr Met Lys Gly Thr Thr Tyr Arg Glu Phe Glu Ser Trp Ile
275 280 285
Asp Gly Cys Lys Asn Cys Thr Cys L8u Asn Gly Thr Ile Gin Cys Glu
290 295 300
Thr Leu Ile Cys Pro Asn Pro Asp Cys Pro Lsu Lys Ser Ala Leu Ala
305 310 315 320
Tyr Val Asp Gly Lys Cys Cys Lys Glu Cys Lys Ser Ile Cys Gin Phe
325 330 335
Gin Gly Arg Thr Tyr Phe Glu Gly Glu Arg Asn Z7w Va1 Tyr Ser Ser
340 345 350
Ser Gly Val Cys Val Leu Tyr Glu Cys Lys Asp Gin Thr Met Lys Leu
355 360 365
Val Glu Ser Ser Gly Cys Pro Ala Leu Asp Cys Pro Glu Ser His Gln
370 375 380
Ile Thr Leu Ser His Ser Cys Cys Lys Val Cys Lys Gly Tyr Asp Phe
385 390 395 400
Cys Ser Glu Arg His Asn Cys Met Glu Asn Ser Ile Cys Arg Asn Leu
405 410 415
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Asn Asp Arg Ala Val Cys Ser Cys Arg Asp Gly Phe Arg Ala Leu Arg
420 425 430
Glu Asp Asn Ala Tyr Cys Glu Asp Ile Asp Glu Cys Ala Glu Gly Arg
435 440 445
His Tyr Cys Arg Glu Asn Thr Met Cys Val Asn Thr Pro Gly Ser Phe
450 455 460
Met Cys Ile Cys Lys Thr Gly Tyr Ile Arg Ile Asp Asp Tyr Ser Cys
465 470 475 480
Thr Glu His Asp Glu Cys Ile Thr Asn Gln His Asn Cys Asp Glu Asn
485 490 495
Ala Leu Cys Phe Asn Thr Val Gly Gly His Asn Cys Val Cys Lys Pro
500 505 510
Gly Tyr Thr Gly Asn Gly Thr Thr Cys Lys Ala Phe Cys Lys Asp Gly
515 520 525
Cys A.rg Asn Gly Gly Ala Cys Ile Ala Ala Asn Val Cys Ala Cys Pro
530 535 540
Gln Gly Phe Thr Gly Pro Ser Cys Glu Thr Asp Ile Asp Glu Cys Ser
545 550 555 560
Asp Gly Phe Val Gin Cys Asp Ser Arg Ala Asn Cys Ile Asn Leu Pro
565 570 575
Gly Trp Tyr His Cys Glu Cys Arg Asp Gly Tyr His Asp Asn Gly Met
580 585 590
Phe Ser Pro Ser Gly Glu Ser Cys Glu Asp Ile Asp Glu Cys Gly Thr
595 600 605
Gly Arg His Ser Cys Ala Asn Asp Thr Ile Cys Phe Asn Leu Asp Gly
610 615 620
Gly Tyr Asp Cys Arg Cys Pro His Gly Lys Asn Cys Thr Gly Asp Cys
625 630 635 640
Ile His Asp Gly Lys Val Lys His Asn Gly Gln Ile Trp Val Leu Glu
645 650 655
Asn Asp Arg Cys Ser Val. Cys Ser Cys Gin Asn Gly Phe Val Met Cys
660 665 670
Arg Arg Met Val Cys Asp Cys Glu Asn Pro Thr Val Asp Leu Phe Cys
675 680 685
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Cys Pro Glu Cys Asp Pro Arg Leu Ser Ser Gln Cys Leu His Gin Asn
690 695 700
Gly Glu Thr Leu Tyr Asn Ser Gly Asp Thr Trp Val Gln Asn Cys Gln
705 710 715 720
Gin Cys Arg Cys Leu: Gln Gly Glu Val Asp Cys Trp Pro Leu Pro Cys
725 730 735
Pro Asp Val Glu Cys Glu Phe Ser Ile Lieu Pro Glu Asn Glu Cys Cys
740 745 750
Pro Arg Cys Val Tthr Asp Pro Cys Gln Ala Asp Thr Ile Arg Asn Asp
755 760 765
Ile Thr Lys Thr Cys Leu Asp Glu Met Asn Val Val Arg Phe Thr Gly
770 775 780
Ser Ser Trp Ile Lys His Gly Tthr Giu Cys Thr Lau Cys Gln Cys Lys
785 790 795 800
Asn Gly His Ile Cys Cys Ser Val Asp Pro Gln Cys Lau Gln Glu Leu
805 810 815
(2) INFORMATION FOR SEQ ID N0:38:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2448 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:
ATGGAG'IC;TC GGGTC.'TTACT GAGAACATTC !'TGATCT TCGGPC,'I'CX~G AGCAG'I=IRGG 60
GGGCTTGGTG TGGACOCTTC CCTACAGATT GACGTC'TTAA CAGAG2TAGA ACT'I'GGGGAG 120
TOCAOGACCG GAG`1`GOGICA GG'ITOCX."GOGG Ct'OC'ATAATG Gt'10GAAAGC CtPI'C'iCTT'P
180
C.AAGATAC'I'C CCAGAAGC;AT AAAAGC'AZTOC ACl'OCTACAG CTGAAC',AU'TT TTTTCAGAAG
240
CnGAGAAATA AACATGAATT TACTATT'ITG GTGAQOC'TAA AACAIGAOOCA CTTAAATTCA 300
GGAGITAZTC TCPC'AATTCA CCACTIGC'aAT CACAQGTAOC TGGAAC'iGGA AAGPAG'i'OC',C 360
CA 02458872 2004-03-23
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CATCGGAATG AAGfrAGACP GCATTAOCGC TC,AGGCAGIC ACOCGCCCPCA CACAGAAGIG 420
'ITL'CJCZTACA TT'PPOOCTGA ZGACAAGPGG CACAAGCTCT CClTAGCCAT CAGPOC'I'PCC 480
CATTTGATTT TACACATTGA CTOCAATAAA ATTTAZGAAA GGGPAGTAGA AAAOOCCI'CC 540
ACAGACT'= CTCPAGG(',AC AACATTTPGG CTAGGP-CA~'~A GAAATAATGC GCATGGATAT 600
TTTAAGGGTA TAATOCAAGA TGTIX',AATTA CiTGPCATGC CCCAGGGATT TATTGCPCAG 660
TGOCCAGATC TTAATCOCAC CIG'i'CCAACT TGCAAZGoACP TCCATGGAGT TGl'GCAGAAA 720
ATCATGGAGC TACAGGATAT TTTAOCCAAA AlCATCAGCCA AGCTG't'C'I'CG AGCTGAACAG 780
C7GAATGAATA GATi'GGATCA GZC~CPATT~I' GAAAGGGAC1T GCACCATGAA GGGAACX'.AICC 840
TAC0C'~AGAAT TTG~QI'CCPG GATAGAOGOC TOtAAGAACT GCAC'ATGOCY GAAZGGAACC 900
ATCX;AQI'('I'G 14P-AL`I'CYAAT C'I'GCCGAAAT aCTt'aACIGCC CACZTAAG'i'C
GGCI'CTTGCG 960
TATGPGGATG GCAAATG'CIG TAAG~'~AATGC AAATCGATAT GCG1ATTi'CA AGGACGAACC 1020
TACTZ'PGAAG GAGAAACAAA TACAGTCTAT TCCZCTTL`IG C'~AGTATGI'GP TCTCPATGAG 1080
TGCAAGGACC AGACCATGAA A(,'T1GT'IGAG AQP1"C,AGOC,`T CZ'OC.AGCTIT GGATIGPOCA
1140
GAG'rCTCATC AGATAAOGZT GTCfCACAGC ZGPI'GC'AAAG TI'TGTAAAQG TTATGACTTT 1200
ZYT=).'CPC',AAA GOC'ATAACPG CATGGAGAAT TCCAZCZGCA GAAAZCT'C'~A TGACAGGGCT 1260
GTTPGrAGCT GPOGAGATGG TTTTAGGGGT CITCGAGAGG ATAATGCCPA CTG!'C'aAAGAC 1320
ATZ7GATGAGr GZGC'i'GAAGG GCGCCATTAC TG'ICJGICAAA ATACAATGTG TGPCAACACC 1380
CCGGGi'I'CTT TTATGI'CCAT CI'OC'AAP,ACT GGATACATCA GAATTGATGA TTATTCATGT 1440
ACAGA,ACATG A'1C'~AGZGTAT CAGAAATCAG CACAAG'TGTG ATGAAAATGC TTTATOCPI'C 1500
AAC'AC't"G'!TG GADGACACAA CTGIGTrTTGC AAGCCGGGCT ATACAGGGAA TIXGAACX"aACA 1560
TGCAP-AGC'.AT TTTGCAAP,GA TCGCTGTAGG AATGGAGGAG CCTG'PATTaC CGCTAATG'IG 1620
TG'I'GCC'i'Of'.C CACAAGGCTT CACTGGACCC AOC'I'GPGAAA CGGACATTGA TGAATOGTCT 1680
GATGGP'i"I"PG TTC'AATGt'GA CAGIC7GTGC.'T AATTGCATTA ACCPGGC'iG1G ATGG'TACCAC
1740
TGTGAGTGCA GAGATGGCTA CCATGACAAT GGGATGITIT CACCAAGi'GG AGAATCGI'GT 1800
GAAGATATTG ATGAGTGPGG GACOOGGAGG CACAGCI'GZG CCAATGATAC CATT'IGGTI'C 1860
CA 02458872 2004-03-23
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AATPTGGATG GOOC'aATATGA TTGTCC;ATGT dC:'I'CATGGAA AGAATTGCAIC AGCGGACTGC 1920
ATC7CAZCATG GAAAP,GPTAA CCAC',AATGGP CAGA-TT'1'GGG TGTIGC'~AP,AA TGACAGGTGC
1980
TCTGZGTCI;T CATGTCAGAA TGGAT'I'CGiT ATGPGPOGAC GGATGGTCTG TGAC7IGZC'aAG 2040
AATCCCACAG TTGAZCTTTr TTG~,'i'GCC7(.~' GAAZGTGACC CAAGGCTTAG TAGPCAGTGC 2100
C't'CC'.ATCAAA ATGGGGAAAC TI'TGTATAP-C AGrGG7GACA CCTGGGPtXA GAATTGICAA 2160
CAGTGOCGCP GCPPGCAAGG GGAAGT'iGAT TGTMGCCCC TGOCTIC~C7CC AGATGTGGAG 2220
'PG'IGAATTCA GCATI'CTOOC AGAGAATGAIG TGC.'TGOff)OGC GCTGIC~PC'.AiC
Af'õAiC1CCTIGC 2280
CAGGC.'TGiACA CCA't'CCGCAA ZGACAT(',ACC AAGACI'POOC TGGACGAAAT GAATG'POG'IT
2340
OGC.'TTC',AOOG WPOCrCPIG GATCAAACAT GGCACPGAGr GTACTCI'C`i'G OCAGTOCAAG 2400
AA'I'OGCC'.ACA TCTGITGCTC AGZGC'~ATCCA C~IGTGCCT'I'C AGGAACTG 2448
(2) INFORMATION FOR SEQ ID NO:39:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 3198 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-093E05
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 97..2544
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:
TTOG("~AGCAG CAGPCTCi'CC GCPOGZC.'ZCC CGGAGGTT'I'C ZC('~ATTGi'CT CTGCCTI'TAC
60
AACAGAGGGA GAOGATGGAC TGAGC."I'GATC CGCACC ATG GAG TCT COG GTC TTA 114
Met Glu Ser Arg Val Leu
CA 02458872 2004-03-23
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1 5
CTG AGA ACA TTC ZGT TTG ATC TPC GGT CTC GGA GCA GTT TGG GGG G'TT 162
Leu Arg Thr Phe Cys Leu Ile Phe Gly Leu Gly Ala Val Trp Gly Leu
15 20
GGT GTG GAC OCT TCC CTA CAG ATT GAC GTC TTA ACA GAG TTA GAA CIT 210
Gly Val Asp Pro Ser Leu Gln Ile Asp Val Leu Thr Glu Leu Glu Lau
25 30 35
GGG GAG TCC AAG AOC GGA GTG WT CAG GTC OOG GflG CTG CAT AAT GGG 258
Gly Glu Ser Thr Thr Gly Val Arg Gln Val Pro Gly Leu His Asn Gly
40 45 50
ACG AAA GOC TTT CIC TTT CAA GAT ACT COC AGA AGC ATA AAA GCA TCC 306
Thr Lys Ala Phe Leu Phe Gin Asp Thr Pro Arg Ser Ile Lys Ala Ser
55 60 65 70
ACT GL'T ACA GCT GAA CAG TTT TTT CAG AAIG CTG AGA AAT AAA CAT GAA 354
Thr Ala Thr Ala Glu Gln Phe Phe Gln Lys Leu Arg Asn Lys His Glu
75 80 85
TTT ACT ATT TTG GTG AOC CTA AAA CAG AOC CAC TTA AAT TCA GGA GTT 402
Phe Thr Ile Leu Val Thr LSu Lys Gln Thr His Leu Asn Ser Gly Val
90 95 100
ATT CTC TCA ATT CAC CAC TTG GAT CAC AGG TAC CZG GAA CZ'G GAA AG'T 450
Ile Leu Ser Ile His His Leu Asp His Arg Tyr Leu Glu Leu Glu Ser
105 110 115
AGT GGC CAT aGG AAT GAA GTC AGA C'I'G CAT TAC OGC TGA GGC AGT CAC 498
Ser Gly His Arg Asn Glu Val Arg Leu His Tyr Arg Ser Gly Ser His
120 125 130
CGC OGT CAC ACA GAA GTG TRT CCT TAC ATT Ti'G GCT GAT GAC AAG TGG 546
Arg Pro His Thr Glu Val Phe Pro Tyr Ile Leu Ala Asp Asp Lys Trp
135 140 145 150
CAC AAG CTC TCC TTA GCC ATC AG'P GCT TCC CAT TTG ATT TTA CAC ATT 594
His Lys Leu Ser Leu Ala Ile Ser Ala Ser His Leu Ile Leu His Ile
155 160 165
GAC TGC AAT AAA ATT TAT GAA AGG GTA GTA GAA AAG OOC TCC ACA GAC 642
Asp Cys Asn Lys Ile Tyr Glu Arg Val Val Glu Lys Pro Ser Thr Asp
170 175 180
TTG (7C'I' CTA GGC ACA ACA T'1'T TaG CTA GGA CAG AGA AAT AAT GGG CAT 690
Leu Pro Leu Gly Thr Thr Phe Trp Leu Gly Gln Arg Asn Asn Ala His
185 190 195
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GGA TAT T'IT AAG GGT ATA AZG CAA GAT GTC CAA TTA CTT GTC ATG MC 738
Gly Tyr Pitie Lys Gly Ile Met Gln Asp Val Gln Leu Leu Val Met Pro
200 205 210
CAG GGA TTT ATT GCT CAG ZGC CCA GAT C.'TI' AAT tJGC ACC TGT CCA AC.T 786
Gin Gly Phe Ile Ala Gln Cys Pro Asp Leu Asn Arg Thr Cys Pro Thr
215 220 225 230
TM AAT GAC TTC CAT GGA CIT GTG CAG AAA ATC ATG GAG CTA CAG GAT 834
Cys Asn Asp Phe His Gly Leu Val Gin Lys Ile Met Glu Leu Gln Asp
235 240 245
ATT TTA QOC AAA ACA TCA GOC AAG CPG TZ.T OGA GCT GAA CAG GGA ATG 882
Ile Leu Ala Lys Thr Ser Ala Lys Leu Ser Arg Ala Glu Gln Arg Met
250 255 260
AAT AGA TTG GAT CAG TGC TAT TGT GAA AGG AGT TGC AOC ATG AAG GGA 930
Asn Arg Leu Asp Gln Cys Tyr Cys Glu Arg Thr Cys Thr Met Lys Gly
265 270 275
AOC AIOC TAC dGA GAA TTT GAG TCC TGG ATA GAC GGC TGT AAG AAC TGC 978
Thr Thr Tyr Arg Glu Rie Glu Ser Trp Ile Asp Gly Cys Lys Asn Cys
280 285 290
ACA TGC CTG AAT GGA ACC ATC CAG TGT GAA ACT CTA ATC TGC CCA AAT 1026
Thr Cys Leu Asn Gly Tthr Ile Gln Cys Glu Thr Leu Ile Cys Pro Asn
295 300 305 310
OL'I' GAC TGC CX'A CTT AAG TCG Gt,"T CIT GCG TAT GTG GAT GGC AAA TGC 1074
Pro Asp Cys Pro Leu Lys Ser Ala ILeu Ala Tyr Val Asp Gly Lys Cys
315 320 325
TGT AAG GAA TGC AAA TM ATA TaC CAA TTT CAA GGA OGA AQC TAC TTT 1122
Cys Lys Glu Cys Lys Ser Ile Cys Gln Phe Gln Gly Arg Thr Tyr Phe
330 335 340
GAA GGA GAA AGA AAT ACA GTC TAT TQC TGT TGY' GGA GTA TGT GTT CrC 1170
Glu Gly Glu Arg Asn Thr Val Tyr Ser Ser Ser Gly Val Cys Val Leu
345 350 355
TAT GAG TGC AAG GAC CAG AOC ATG AAA CIT GTT GAG AGT TGA GGC TGT 1218
Tyr Glu Cys Lys Asp Gln Thr Met Lys Leu Val. Glu Ser Ser Gly Cys
360 365 370
OCA GCT TIG GAT TGT UCA GAG TCT CAT CAG ATA ACC TTG TCT CAC AGC 1266
Pro Ala Leu Asp Cys Pro Glu Ser His Gln Ile Thr Leu Ser His Ser
375 380 385 390
TG'P TGC AAA GTT TGT AAA GGT TAT GAC TTT TGT TCT GAA AGG CAT AAC 1314
Cys Cys Lys Val qis Lys Gly Tyr Asp Phe Cys Ser Glu Arg His Asn
CA 02458872 2004-03-23
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395 400 405
TW ATG GAG AAT TOC ATC TGC AGA AAT C'IG AAT GAC AGG GCT GTP ZGT 1362
Cys Met Glu Asn Ser Ile Cys Arg Asn LSu Asn Asp Arg Ala Val Cys
410 415 420
AGC TGT CGA GAT GGT TTT AGG GGT C'I'I' CGA GAG GAT AAT GOC TAC TGT 1410
Ser Cys Arg Asp Gly Phe Arg Ala Leu Arg Glu Asp Asn Ala Tyr Cys
425 430 435
GAA GAC ATC GAT GAG TGT GCT GAA GOG OOC CAT TAC TGT CGT GAA AAT 1458
Glu Asp Ile Asp Glu Cys Ala Glu Gly Arg His Tyr Cys Arg Glu Asn
440 445 450
ACA ATG TGT GTC AAC AOC OCG GGT TCT TTT ATG TGC ATC TGC AAA ACT 1506
Thr Met Cys Val Asn Thr Pro Gly Ser Ptye Met Gys Ile Cys Lys Thr
455 460 465 470
GGA TAC ATC AGA ATT GAT GAT TAT TCA TGT ACA GAA CAT GAT GAG TGT 1554
Gly Tyr Ile Arg Ile Asp Asp Tyr Ser Cys Thr Glu His Asp Glu Cys
475 480 485
ATC ACA AAT CAG CAC AAC TGT GAT GAA AAT GCT TTA TGC TTC AAC AIGT 1602
Ile Thr Asn Gin His Asn Cys Asp Glu Asn Ala Lau Cys Phe Asn Thr
490 495 500
GTT GGA GGA CAC AAC TGT GTT TGC AAG OCG GGC TAT ACA GGG AAT GGA 1650
Val Gly Gly His Asn Cys Val Cys Lys Pro Gly Tyr Thr Giy Asn Gly
505 510 515
AOG ACA TOC AAA GCA TTT TGC AAA GAT GGC TGT AGG AAT GGA GGA GOC 1698
Thr Thr Cys Lys Ala Phe Cys Lys Asp Gly Cys Arg Asn Gly Gly Ala
520 525 530
TGT ATT GOC GGT AAT GTG TGT GOC TGC CCA CAA GGC TTC AGT GGA OOC 1746
Cys Ile Ala Ala Asn Val Cys Ala Cys Pro Gin Gly Phe Thr Gly Pro
535 540 545 550
AOC TGT GAA AOG GAC ATTT GAT GAA TGC TCP GAT GGT TTT GTT CAA TGT 1794
Ser Cys Glu Thr Asp Ile Asp Glu Cys Ser Asp Gly Phe Val Gln Cys
555 560 565
GAC AGT C:GT GC.T AAT TGC ATT AAC CTG CCT GGA TGG TAC CAC TGT GAG 1842
Asp Ser Arg Ala Asn Cys Ile Asn Leu Pro Gly Trp Tyr His Cys Glu
570 575 580
TOC AGA GAT GGC TAC CAT GAC AAT GGG ATG TTT ZCA CCA AGT GGA GAA 1890
Cys Arg Asp Gly Tyr His Asp Asn Gly Met Phe Ser Pro Ser Gly Glu
585 590 595
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TCG TGT GAA GAT ATT GAT GAG TGT GGG AOC GGG AGG CAC AGC TGT GM 1938
Ser Cys Glu Asp Ile Asp Glu Cys Gly Thr Gly Arg His Ser Cys Ala
600 605 610
AAT GP-T ACC ATT TGC TTC AAT TTG GAT GGO GGA TAT GP,T TGT (7GA TG'I' 1986
Asn Asp Thr Ile Cys Phe Asn Leu Asp Gly Gly Tyr Asp Cys Arg Cys
615 620 625 630
OCT CAT GGA AAG AAT TGC ACA GGG GAC TGC ATC CAT GAT GGA AAA GTT 2034
Pro His Gly Lys Asn Cys Thr Gly Asp Cys Ile His Asp Gly Lys Val
635 640 645
AAG CAC AAT WT CAG ATT TGG GTG T1'G G;AA AAT GAC AGG TGC TG'T GTG 2082
Lys His Asn Gly Gln Ile Trp V&l. ieu Glu Asn Asp Arg Cys Ser Val
650 655 660
TGC TCA TGT CAG AAT GGA TTC GTT ATG TGT OGN OGG ATG GTC TGT GAC 2130
Cys Ser Cys Gln Asn Gly Phe Val Met Cys Arg Arg Met Val Cys Asp
665 670 675
TGT GAG AAT GOC ACA GTT GAT CTT TTT TGC TOC aCT GAA TGT GAC OCA 2178
qs Glu Asn Pro Thr Val. Asp Leu Phe Cys Cys Pro Glu Cys Asp Pro
680 685 690
AGG CTT AGT AGT CAG TGC CTC CAT CAA AAT GGG GAA ACT TTG TAT AAiC 2226
Arg Leu Ser Ser Gln Cys Leu His Gln Asn Gly Glu Thr Leu Tyr Asn
695 700 705 710
AGT GGT GAC ACC TGG GTC CAG AAT TGT CAA CAG TGC C:OC TGC TTG CAA 2274
Ser Gly Asp Thr Trp Val Gln Asn Cys Gln Gln Cys Arg Cys Leu Gln
715 720 725
GGG GAA GTT GAT ZGT TGG COC CIG OGT TGC CCA GAT GTG GAG TGT GAA 2322
Gly Glu Val Asp Cys Trp Pro Leu Pro Cys Pro Asp Val Glu Cys Glu
730 735 740
TTC AOC ATT CTC OCA GAG AAT GAG TGC TGC QOG aGC TGT GTG ACA GAC 2370
Phe Ser Ile Leu Pro Glu Asn Glu Cys Cys Pro Arg Cys Val Thr Asp
745 750 755
OCT TaC CAG GCT GAC ACC ATC aCC AAT GAC ATC ACC AAG ACT TGC CIG 2418
Pro Cys Gln Ala Asp Thr Ile Arg Asn Asp Ile Thr Lys Thr Cys Leu
760 765 770
GAC GAA ATG AAT GZG GTT OOC TTC A(JC GOG TOC TCT TC3G ATC AAA CAT 2466
Asp Glu Met Asn Val Val Arg Phe Thr Gly Ser Ser Trp Ile Lys His
775 780 785 790
GOC ACT GAG TGT ACT CIC TGC CAG TGC AP-G AAT GGC CAC ATC TGP TGC 2514
Gly Thr Glu Cys Thr Leu Cys Gln Cys Lys Asn Gly His Ile Cys Cys
CA 02458872 2004-03-23
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795 800 805
TCA GTG GAT CCA CAG TGC C'IT CAG GAA CIG ZGAAGT'rAAC TG'IC'I'CATGG 2564
Ser Val Asp Pro Gin Cys Leu Gln Glu Leu
810 815
GAGATTZC,'IG ZTAAAAGAAT GTTCTITCAT TAAAAGAOCA AAAAGAAG'TT AAAACTTAAA 2624
TTG(~GI'GATT TGi'GGGCA10C TAAATGCAGC TTI'G`ITAATA GCTGAGIY'yAA CTT`1'C'.AATTA
2684
ZGAAAZ'I'IGr GGAGCTPC',ArC AAAATCA('AA AAQC''AAAATT ACTGGGGCAA AATTACACGT 2744
CAAG'I'C7I'GOC TCTArCZC3TGT CTCACATCAC CATGTAGAAG AATGGGCGTA CAGI'ATATAC 2804
CGTGACATCC TGAAOCL'TOG ATAGAAADCJC TGAOOOCATT GGATCTG'IGA AAGCC,R'CTAG 2864
CT1C'.A~.'iC~T GCAC'aAAAATT TPOC'!'CI'AGA TC'AGAATLZ'T C.AC'aAATCAGT
TA~OGTTOCIC 2924
ACTGC.AAGAA ATAAAATGIC AGGC=AGI'GAA TGAATTATAT TTTCAGAAGr AAAGCAAA('1 2984
AGCTATAACA TGTTATGTAC AIGTACAC'PG'P G1?AAAGAAAT Ci''aAAAC.AAC' TTATTGTAAT 3044
GATAAAAATA ATGCACAGOC ATG(7ITAC'iT AATATTTTCP AACAOGAAAA GTCATCOC'PA 3104
TTI'IOCTI'GZT TTAC'iYGC.ACT TAATATTATT TGGT'I'GAATT TGTTC.AGPAT AAGCTCGTIC
3164
TTC'IG('.AAAA TTAAATAAAT ATT'i~CICTTA CC'iT 3198
(2) INFORMATION FOR SEQ ID N0:40:
(i) SEQUENCE.CHARACTERISTICS:
(A) LENGTH: 499 amino acids
(B) TYPE: amino acid
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:
Met Glu Leu Ser Glu Pro Val Val Glu Asn Gly Glu Val Glu Met Ala
1 5 10 15
Leu Glu Glu Ser Trp Glu His Ser Lys Glu Val Ser Glu Ala Glu Pro
20 25 30
Gly Gly Gly Ser Ser Gly Asp Ser Gly Pro Pro Glu Glu Ser Gly Gln
35 40 45
CA 02458872 2004-03-23
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Glu Met Met Glu Glu Lys Glu Glu Ile Arg Lys Ser Lys Ser Val Ile
50 55 60
Val Pro Ser Gly Ala Pro Lys Lys Glu His Val Asn Val Val Phe I1e
65 70 75 80
Gly His Val Asp Ala Gly Lys Ser Thr Ile Gly Gly Gln Ile Met Phe
85 90 95
Leu Thr Gly Met Ala Asp Lys Arg Thr Lau Glu Lys Tyr Glu Arg Glu
100 105 110
Ala Glu Glu Lys Asn Arg Glu Tthr Trp Tyr Leu Ser Trp Ala Leu Asp
115 120 125
Thr Asn Gln Glu Glu Arg Asp Lys Gly Lys Thr Val Glu Val Gly Arg
130 135 140
Ala Tyr Phe Glu Thr Glu Arg Lys His Phe Thr Ile Leu Asp Ala Pro
145 150 155 160
Gly His Lys Ser Phe Val Pro Asn Met Ile Gly Gly Ala Ser Gln Ala
165 170 175
Asp Leu Ala Val Leu Val Ile Ser Ala Arg Lys Gly Glu Phe Glu Thr
180 185 190
Gly Phe Glu Lys Gly Gly Gln Thr Arg Glu His Ala Met Phe Gly Lys
195 200 205
Thr Ala Gly Val Lys His Leu Ile Val Lau Ile Asn Lys Met Asp Asp
210 215 220
Pro Thr Val Asn Trp Gly Ile Glu Arg Tyr Glu Glu Cys Lys Glu Lys
225 230 235 240
Leu Val Pro Phe Lau Lys Lys Val Gly Phe Ser Pro Lys Lys Asp Ile
245 250 255
His Phe Met Pro Cys Ser Gly Lau Thr Gly Ala Asn Ile Lys Glu Gln
260 265 270.
Ser Asp Phe Cys Pro Trp Tyr Thr Gly Lsu Pro Phe Ile Pro Tyr Lsu
. 275 280 285
Asn Asn Lsu Pro Asn Phe Asn Arg Ser Ile Asp Gly Pro Ile Arg Lau
290 295 300
Pro Ile Val Asp Lys Tyr Lys Asp Met Gly Thr Val Val Leu Gly Lys
305 310 315 320
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Leu Glu Ser Gly Ser Ile Phe Lys Gly Gln Gln Leu Val Met Met Pro
325 330 335
Asn Lys His Asn Val Glu Val Leu Gly Ile Leu Ser Asp Asp Thr Glu
340 345 350
Thr Asp Phe Val Ala Pro Gly Glu Asn Leu Lys Ile Arg Leu Lys Gly
355 360 365
Ile Glu- Glu Glu Glu Ile Leu Pro Glu Phe Ile Leu Cys Asp Pro Ser
370 375 380
Asn Leu Cys His Ser Gly Arg Thr Phe Asp Val Gln Ile Val Ile Ile
385 390 395 400
Glu His Lys Ser Ile Ile Cys Pro Gly Tyr Asn Ala Val Leu His Ile
405 410 415
His Thr Cys Ile Glu Glu Val Glu Ile Thr Ala Leu Ile Ser Leu Val
420 425 430
Asp Lys Lys Ser Gly Glu Lys Ser Lys Thr Arg Pro Arg Phe Val Lys
435 440 445
Gln Asp Gln Val Cys Ile Ala Arg Leu Arg Thr Ala Gly Thr Ile Cys
450 455 460
Leu Glu Thr Phe Lys Asp Phe Pro Gln Met Gly Arg Phe Thr Leu Arg
465 470 475 480
Asp Glu Gly Lys Thr Ile Ala Ile Gly Lys Val Leu Lys LSu Val Pro
485 490 495
Glu Lys Asp
(2) INFORMATION FOR SEQ ID NO:41:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1497 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(xi) SEQUENCE DESCRIPTION: SEQ ID N0:41:
AT'GGAACTTT CAGAP-OCTGT TGTACAAP-AT GGAIGAGGI'GG AAATGOIXX.T AGAAGAATCA 60
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TGGGAGCACA GTAAAGAAG'P AAGT('~AAQOC GAGCCTOC'GG GI'OG'I"IC7C'PC GGGAGATTCA 120
OGGCCCCCAG AAGAAAGZ'OG CCAaGAAATG ATGGAGGAAA AAGAOGAAAT AA~C'aAAAATCC 180
AAATCTGTGA TCGPACCC'I'C AGGTGCACCT AAC''AAAGAAC ACGI'AAATGT AGPATTCATT 240
GGCX'.ATGTAG ACGC,'ZCGCAA GTCAACCATC GGAIGGACAGA TAATGTPTIT GACTGGAATG 300
GCTGACAAAA GAACACTOGA GAAATATC'aAA AGAGAAGCTG AOGAAAAAAA CAG:AC'~AACC 360
ZTATGT 1C1 T AGATAC,AAAT CAGC',oAGGAAC GAGACAAGGG TAAAACAGIC 420
GAAGTGGI'C GTGCC.'TATTT TGAAACAGAA AOGAAAICATT TCACAATTTI' AGATGCOCC,T 480
COGCCACAAGik GTTTT(~i'OCC AAP-TA'I'GATT uG1wI'C~CIT C'iC',AAGCi'GA
TPICaGCi~GIC` 540
CTaGTCATCT CTGCCAIGGAA AGCAGAGTTT GAAAC,'TGGAT TTGAAAAAOG TOGACAGACA 600
AGAC'aAAf'.ATG CGAZGrrTGG CAAAACOGC',A GGAGPAAAAIG ATTTAATAGr GCI'TATTAAT 660
AAGATGGATG ATCCCACAGP AAATTGOGGC ATCXGAGAGAT ATGAAGAATG TAAAOAAAAA 720
C'IC~GI'OOOC.'T TTTTGAAAAA AGTA GCPIT AQi'CCAAAAA AGGACATZCA CTTTA'IG:CC 780
TGCPC'AOGAC 'i'GACCGGAGC AAATATTAAA GAGCAGICAG ATTTCTGOCC TZGCZ'ACACr' 840
GGATTACCAT TTATTCCGrA TTTt3AATAAC TTGOC'AAACT TCAACA[3ATC AATTGATGGA 900
CCAATAAGAC TGCf'.AATTGT GGATAAOTAC AAAiGATATOG OCAGTGTGGT CCTGGGAAPdG 960
CPCGAAT= GGTCX".ATTTT TAAAGGCCAG CAGCTCGTGA TGATOCCAAA CAAGCACAAT 1020
GTAlGAAGITC TTGC'~AATACT TTCTC'rATGAT ACZGAAAL.'TG ATI"I'I'GTAOC CCC'.AIOGrGAA
1080
AAOCTCAAAA TCAGACTGAA G'OGAATIGAA GAAGAAGAC'aF- TTCZ'PCC1AC'aA ATTCATACZT 1140
TGTGATCCrA GTAACCI'CTG CCAT'PC'PGGA COCACGITIG ATGTIC'Af'aAT AGi'GATTATT 1200
GACY'A,C'AAAT CCATCATCTG CCCAGGT'rAT AATGCGGTaC TCGCACATTCA TAGTTGTATT 1260
C;AGGAAGT'iG AGATAACAGC GTTAATCTCC TTOGTAGAf'A AAAAAT('.AOG aGAAAAAAIG'T 1320
AAGACACGAC CCIX'CTPOGT GAAAOAAOAT CAAGrATGCA TT{~CrCG!'t'T AAGGACAGCA 1380
GGAAOCATCT GOCI'CGAGAC GPTC,AAAGAT TTTCCPCAGA TOOGTOGI'IT TACTTTAAGA 1440
GATGAGGGTA AGACCATTGC AATTG('aAAAA G'I'I'CTGAAAT TGGTCCCAGA GAAGGAC 1497
(2) INFORMATION FOR SEQ ID N0:42:
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(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 2057 base pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: DNA(genomic)
(iii) HYPOTHETICAL: NO
(iv) ANTI-SENSE: NO
( vii ) IMMEDIATE SOURCE:
(A) LIBRARY: Human fetal brain cDNA library
(B) CLONE: GEN-077A09
(ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION: 144..1640
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:
Ta00GGCCOG CTCOGGC'.AGC AAL7C`AZGW OCrOCACCGG CGCGOf'aATAIC aCTCAA[OGTA 60
AAAGGATaGG ACGGGGGGCA CC,`I'G7GGAAC CIZC00f'AGA GGAAOOGTTA GiG'POOClTG 120
AAOGPIMAA Ti'C.AGOOGPT ACC ATG GAA CT'T TCA GAA CCT GTT GTA GAA 170
Met Glu Leu Ser Glu Pro Val Val Glu
1 5
AAT GGA GAG GTG GAA ATG GOC CTA GAA GAA TCA TGG GAG CAC AGT AAA 218
Asn Gly Glu Val Glu Met Ala Leu Glu Glu Ser Txp Glu His Ser Lys
15 20 25
GAA GTA AGT GAA GCC GAG CGT GGG GGT GGT TCC TCG GGA GAT TCA GGG 266
Glu Val Ser Glu Ala Glu Pro Gly Gly Gly Ser Ser Gly Asp Ser Gly
30 35 40
CCC CCA GAA GAA AGT GGC CAG GAA ATG ATG GAG GAA AAA GAG GAA ATA 314
Pro Pro Glu Glu Ser Gly Gln Glu Met Met Glu Glu Lys Glu Glu Ile
45 50 55
AGA AAA TCC AAA TCT GTG ATC GTA CCC TCA GGT GCA CCT AAG AAA GAA 362
Arg Lys Ser Lys Ser Val Ile Val Pro Ser Gly Ala Pro Lys Lys Glu
60 65 70
CAC GTA AAT GTA GTA TTC ATT GGC CAT GTA GAC GCT GGC AAG TCA ACC 410
His Val Asn Val Val Phe Ile Gly His Val Asp Ala Gly Lys Ser Thr
75 80 85
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ATC GGA GGA CAG ATA ATG TTT TTG ACT GGA ATG GCT GAC AAA AGA ACA 458
Ile Gly Gly Gln Ile Met Phe L8u Thr Gly Met Ala Asp Lys Arg Thr
90 95 100 105
CTG GAG AAA TAT GAA AGA GAA GCT GAG GAA AAA AAC AGA GAA ACC ZGG 506
Leu Glu Lys Tyr Glu Arg Glu Ala Glu Glu Lys Asn Arg Glu Thr Trp
110 115 120
TAT Tt'G TCC TGG GOC TTA GAT ACA AAT CAG GAG GAA CGA GAC AAG GGT 554
Tyr Leu Ser Trp Ala Leu Asp Thr Asn Gln Glu Glu Arg Asp Lys Gly
125 130 135
AAA ACA GTC GAA GTG GGT CGT GCC TAT TTT GAA ACA GAA AGG AAA CAT 602
Lys Thr Val Glu Val Gly Arg Ala Tyr Phe Glu Thr Glu Arg Lys His
140 145 150
TTC ACA ATT TPA GAT GflC CCT GGC CAC AAG AGT TTT GTC CCA AAT ATG 650
Phe Thr Ile Leu Asp Ala Pro Gly His Lys Ser Phe Val Pro Asn Met
155 160 165
ATT GGr GGT GCT TC'T CAA GCT GAT TTG GLT GTG CIG GIC ATC TCT GOC 698
Ile Gly Gly Ala Ser Gln Ala Asp Leu Ala Val Leu Val Ile Ser Ala
170 175 180 185
AGG AAA GGA GAG TTT GAA ACT GGA TTT GAA AAA GGT GGA CAG ACA AGA 746
Arg Lys Gly Glu Phe Glu Thr Gly Phe Glu Lys Gly Gly Gln Thr Arg
190 195 200
GAA CAT GCG ATG TTT GOC AAA AIOG a'.A GGA G'TA AAA CAT TTA ATA GTG 794
Glu His Ala Met Phe Gly Lys Thr Ala Gly Val Lys His Leu Ile Val
205 210 215
CTT AZT AAT AAG ATG GAT GAT COC ACA GTA AAT T G GOC ATC GAG AGA 842
Leu Ile Asn Lys Met Asp Asp Pro Thr Val Asn Trp Gly Ile Glu Arg
220 225 230
TAT'GAA GAA TGT AAA GAA AAA CIG GIG CCC TTT TTG AAA AAA GTA GGC 890
Tyr Glu Glu Cys Lys Glu Lys Leu Val Pro Phe Leu Lys Lys Val Gly
235 240 245
TTT AGT CCA AAA AAG GAC AZT CAC TTT ATG CCC TGC TCA GGA CTG ACC 938
Phe Ser Pro Lys Lys Asp Ile His Phe Met Pro Cys Ser Gly Leu Thr
250 255 260 265
GGA GCA AAT ATT AAA GAG CAG 'ICA GAT TTC TGC CCT TOG TAC ACT GGA 986
Gly Ala Asn Ile Lys Glu Gln Ser Asp Phe Cys Pro Trp Tyr Thr Gly
270 275 280
TTA CCA TTT AZT CCG TAT TTG AAT AAC TTG CCA AAC TZC AAC AGA '1CA 1034
Leu Pro Phe Ile Pro Tyr Leu Asn Asn Leu Pro Asn Phe Asn Arg Ser
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285 290 295
ATT GAT GGA Cf_'A ATA AGA C'1G CCA ATT GTG GAT AAG TAC AAA GAT ATG 1082
Ile Asp Gly Pro Ile Arg Leu Pro Ile Val Asp Lys Tyr Lys Asp Met
300 305 310
GOC ACT GTG GTC C't'G GGA AAG C'I'G GAA TOC GGG TOC ATT TTT AP-P- GGC 1130
Gly Thr Val Val Leu Gly Lys Leu Glu Ser Gly Ser Ile Phe Lys Gly
315 320 325
CAG CAG CTC GTG ATG ATG OGA AAIC AAL", CAC AAT GTA GAA GTT C'!T GGA 1178
Gln Gl.n. Leu Val Met Met Pro Asn Lys His Asn Val Glu Val Leu Gly
330 335 340 345
ATA CI'T TCT GAT GAT ACT GAA ACT GAT TTT GTA GCC OCA GGT GAA AAC 1226
Ile Leu Ser Asp Asp Thr Glu Thr Asp Phe Val Ala Pro Gly Glu Asn
350 355 360
CTC AAA ATC AGA C'IG AAIG GGA ATT GAA GAA GAA GAG ATT CIT CCA GAA 1274
Leu Lys Ile Arg Leu Lys Gly Ile Glu Glu Glu Glu Ile Leu Pro Glu
365 370 375
TTC ATA CZT ZGT GAT OCT AGT AAC CTC TGC CAT TCT GGA CGC AOG TTT 1322
Phe Ile Leu Cys Asp Pro Ser Asn Leu Cys His Ser Gly Arg Thr Phe
380 385 390
GAT GTT CAG ATA GTG ATT ATT GAG CAC AAA TCC ATC ATC TGC CCA GGT 1370
Asp Val Gln Ile Val Ile Ile Glu His Lys Ser Ile Ile Cys Pro Gly
395 400 405
TAT AAT GOG GTG CZG CAC ATT CAT ACT TGP ATT GAG GAA GTT GAG ATA 1418
Tyr Asn Ala Val Leu His Ile His Thr Cys Ile Glu Glu Val Glu Ile
410 415 420 425
ACA GOG TTA ATC TOC TTG GTA GAC AAA AAA TCA GGG GAA AAA AGT AAG 1466
Thr Ala Lau Ile Ser Leu Val Asp Lys Lys Ser Gly Glu Lys Ser Lys
430 435 440
ACA OGA OCC OOC TTC GTG AAA CAA GAT CAA GTA TGC ATT GCT CGT TTA 1514
Thr Arg Pro Arg Phe Val Lys Gin Asp Gin Val Cys Ile Ala Arg Leu
445 450 455
AGG ACA GCA GGA AOC ATC TGC CPC GAG ACG TTC AAA GAT TTT C)CT CAG 1562
Arg Thr Ala Gly Thr Ile Cys Leu Glu Thr Phe Lys Asp Phe Pro Gln
460 465 470
ATG GGT CG'I' TTT ACT ZTA AGA GAT GAG GGT AA+G AOC ATT GCA ATT GGA 1610
Met Gly Arg Phe Thr Leu Arg Asp Glu Gly Lys Thr Ile Ala Ile Gly
475 480 485
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AAA GTT CTG AAA TTG GTC aCA GAG AAG GAC TAAGCAATTT TGT'IC'~AZGC.C 1660
Lys Val Leu Lys Leu Val Pro Glu Lys Asp
490 495
TCZGCAAGAT ACTGIC'~AlaGA GAATTGACAG CAAAAGTTCA OCAQCTAC'I'C TTATTTACZG 1720
CCOC'.ATTGATT GACT'ITTCTT CATAT'I'Pl'OC AAAGAGAAAT TTCACAGCAA AAATTCAZGT 1780
TTZGTC'AGCT TTG'PC'ATGTT GAGATCTG'TT A'I'GTCIACZGA TGAATTTAOC CI'CAAGP'i'TC
1840
GTTaCTCTGT AOCACTCTGC TTOCrTGGAC AATATCAGTA ATAGCTTTGT AAGI'GATG'1G 1900
GAOGPAATTG CCTACAU`PAA TAAAAAAATA AT6TACPITA ATPPTPCATT TI'CPT'ITAGG 1960
ATATTTAGAIC CAOOCTrGTT aCAOGC'.AAAC CAGAGTGPGT CAGTG'TTTGT GZ'GTGICTTA 2020
AAATGATAAC TAACATGt'GA ATAAAATACT OCATTTG 2057
