U.S. patent application number 13/565425 was filed with the patent office on 2013-07-11 for identification of modulators of autophagy.
This patent application is currently assigned to RUTGERS, THE STATE UNIVERSITY OF NEW JERSEY. The applicant listed for this patent is Richard Possemato, David M. Sabatini, Anne M. Strohecker, Eileen White. Invention is credited to Richard Possemato, David M. Sabatini, Anne M. Strohecker, Eileen White.
Application Number | 20130178382 13/565425 |
Document ID | / |
Family ID | 42731031 |
Filed Date | 2013-07-11 |
United States Patent
Application |
20130178382 |
Kind Code |
A1 |
White; Eileen ; et
al. |
July 11, 2013 |
Identification of Modulators of Autophagy
Abstract
Methods for identifying compounds that inhibit or stimulate the
autophagy pathway are described. Devices for detecting the
expression of autophagy-related genes and kits for assaying the
expression of autophagy-related genes are also described. Also
described are methods for identifying individuals susceptible to or
afflicted with a disease state associated with an autophagy pathway
defect.
Inventors: |
White; Eileen; (Princeton,
NJ) ; Strohecker; Anne M.; (North Plainfield, NJ)
; Possemato; Richard; (Brighton, MA) ; Sabatini;
David M.; (Cambridge, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
White; Eileen
Strohecker; Anne M.
Possemato; Richard
Sabatini; David M. |
Princeton
North Plainfield
Brighton
Cambridge |
NJ
NJ
MA
MA |
US
US
US
US |
|
|
Assignee: |
RUTGERS, THE STATE UNIVERSITY OF
NEW JERSEY
New Brunswick
NJ
|
Family ID: |
42731031 |
Appl. No.: |
13/565425 |
Filed: |
August 2, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13422033 |
Mar 16, 2012 |
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13565425 |
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13284923 |
Oct 30, 2011 |
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13422033 |
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13046033 |
Mar 11, 2011 |
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13284923 |
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12622410 |
Nov 19, 2009 |
8187802 |
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13046033 |
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61313097 |
Mar 11, 2010 |
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61116085 |
Nov 19, 2008 |
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Current U.S.
Class: |
506/9 ; 435/6.13;
506/10; 506/16; 506/18 |
Current CPC
Class: |
G01N 33/5023 20130101;
C12Q 1/025 20130101; C12Q 1/6809 20130101; G01N 33/53 20130101;
G01N 33/5008 20130101 |
Class at
Publication: |
506/9 ; 435/6.13;
506/16; 506/18; 506/10 |
International
Class: |
C12Q 1/02 20060101
C12Q001/02; G01N 33/53 20060101 G01N033/53; C12Q 1/68 20060101
C12Q001/68 |
Goverment Interests
GOVERNMENT SUPPORT
[0002] The present application was supported in part by the
National Institutes of Health under Grant Nos. R37 CA53370 and RO1
CA130893 and the Department of Defense under DOD W81XWH06-1-0514
and DOD W81XWH05. The U.S. government has certain rights in the
invention.
Claims
1. A method for identifying compounds that inhibit or stimulate the
autophagy pathway for treatment of a disease state associated with
an autophagy pathway defect, comprising measuring the effect of one
or more test compounds on the inhibition or stimulation of a
product of one or more of the genes or gene fragments identified in
Table 3 or Table 4.
2. A method for identifying individuals susceptible to or afflicted
with a disease state associated with an autophagy pathway defect,
comprising testing a biological sample from an individual for a
characteristic of one or more polypeptides produced by expression
of one or more of the genes or gene fragments identified in Table 3
or Table 4 that is indicative of said disease state, wherein said
characteristic is selected from the presence of at least one of
said polypeptides, the absence of at least one of said
polypeptides, an elevated level of at least one of said
polypeptides, a reduced level of at least one of said polypeptides
and, for two or more of said polypeptides, combinations
thereof.
3. A device for detecting the expression of a plurality of
autophagy-related genes associated with an autophagy pathway
defect, said device comprising a substrate to which is affixed at
known locations a plurality of probes, wherein the probes comprise:
a) a plurality of oligonucleotides or polynucleotides, each of
which specifically binds to a different sequence selected from any
of the sequences identified in Table 3 or Table 4 or fragments
thereof; or b) a plurality of polypeptide binding agents, each of
which specifically binds to a different polypeptide or fragment
thereof produced by expression of a gene or gene fragment
comprising any of the sequences identified in Table 3 or Table 4 or
fragments thereof.
4. A kit for assaying the expression of autophagy-related genes
associated with an autophagy pathway defect, comprising at least
one container and a collection of two or more probes, wherein the
probes comprise: a) oligonucleotides or polynucleotides that
specifically bind to two or more genes or gene fragments comprising
any of the sequences identified in Table 3 or Table 4, or fragments
thereof; or b) polypeptide binding agents that specifically bind to
polypeptides produced by expression of two or more genes or gene
fragments comprising any of the sequences identified in Table 3 or
Table 4, or fragments thereof.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a Continuation of U.S. patent
application Ser. No. 13/422,033, filed Mar. 16, 2012, which is a
Continuation of U.S. patent application Ser. No. 13/284,923, filed
Oct. 30, 2011, which is a Continuation of U.S. patent application
Ser. No. 13/046,033, filed Mar. 11, 2011, which claims benefit
under 35 U.S.C. .sctn.119(e) of U.S. Provisional Patent Application
Ser. No. 61/313,097, filed Mar. 11, 2010. U.S. patent application
Ser. No. 13/046,033 is also a Continuation-in-Part of U.S. patent
application Ser. No. 12/622,410, filed Nov. 19, 2009, which claims
benefit under 35 U.S.C. .sctn.119(e) of U.S. Provisional Patent
Application Ser. No. 61/116,085, filed Nov. 19, 2008. The
disclosures of each of the foregoing applications are hereby
incorporated herein by reference in their entireties.
BACKGROUND OF THE INVENTION
[0003] Autophagy is a catabolic, cellular self-digestion process
that is activated by starvation and stress whereby double membrane
vesicles called autophagosomes form that engulf proteins and
organelles. Autophagosomes then fuse with lysosomes where their
cargo is degraded. The function of autophagy is to recycle
intracellular nutrients to sustain metabolism during nutrient and
growth factor deprivation, and to clear damaged proteins and
organelles that accumulate during stress. Although elimination of
individual proteins occurs by the ubiquitin-mediated proteasome
degradation pathway, only the autophagy pathway can eliminate
protein aggregates and organelles. Thus, autophagy complements and
overlaps with proteasome function to prevent the accumulation of
damaged cellular components during starvation and stress. Through
these functions, autophagy is an important cellular stress response
that functions to maintain protein and organelle quality control,
protect the genome from damage, and sustain cell and mammalian
viability.
[0004] Autophagy is controlled by ATG proteins that were initially
identified in yeast for which there are mammalian homologues. ATG
proteins are comprised of kinases, proteases, and two
ubiquitin-like conjugation systems that likely function in concert
with a host of unknown cellular proteins to control autophagosome
formation, cargo recognition, engulfment, and trafficking to
lysosomes. The ATG6/Beclin1-VPS34-ATG8/LC3 complex regulates
autophagosome formation and LC3 cleavage, lipidation, and membrane
translocation are frequently utilized to monitor autophagy
induction and inhibition of flux through the autophagy pathway.
[0005] Targeting of cargo, including proteins and organelles, to
autophagosomes for degradation is accomplished by tagging proteins
with polyubiquitin. The ubiquitin-binding domain (UBA) on the
adaptor protein p62 recognizes and binds these polyubiquitinated
proteins. p62 oligomerizes by self-association of its PB1 domain
and binds ATG8/LC3 on autophagosome membranes. p62 thereby
identifies, collects and delivers cargo to autophagosomes for
degradation. p62 itself is an autophagy substrate and is degraded
by autophagy along with the cargo. As such, p62 accumulation in
aggregates is indicative of autophagy inhibition and clearance of
p62 following stress is indicative of functional autophagy. These
properties of p62 have been demonstrated in vivo in
autophagy-defective mutant mice and are mimicked by expression of
EGFP-p62 in cell lines in vitro and in vivo (Mathew, R et al.,
(2009) Cell 137, 1062-1075).
[0006] The activation of autophagy by starvation and stress is
controlled in part through the PI-3 kinase pathway via the protein
kinase mTOR. Growth factor and nutrient availability promote mTOR
activation that suppresses autophagy, whereas starvation and mTOR
inactivation stimulate autophagy. While there are other mechanisms
to regulate autophagy, and those that activate autophagy in
response to stress are particularly poorly understood, mTOR
provides a link between nutrient and growth factor availability,
growth control, autophagy, and metabolism.
[0007] Autophagy dysfunction is believed to be a major contributor
to human diseases including neurodegeneration, liver disease, and
cancer. Many human neurodegenerative diseases are associated with
aberrant protein accumulation and excessive neuronal cell death,
and neurons of mice with targeted autophagy defects accumulate
polyubiquitinated- and p62-containing protein aggregates that
result in neurodegeneration. The human liver disease
steatohepatitis and a major subset of hepatocellular carcinomas
(HCCs) are associated with the formation of p62-containing protein
aggregates (Mallory bodies), and livers of mice with autophagy
defects have p62-containing protein aggregates, excessive cell
death, and HCC.
[0008] Evidence from model organism disease models indicates that
promoting autophagy with mTOR inhibitors such as rapamycin or
CCI-779, and enhancing the clearance of misfolded, damaged or
mutated proteins and protein aggregates prevents neurodegeneration,
but that there also are mTOR-independent means to increase
autophagy. Similarly, genetically eliminating the expression of p62
in hepatocytes and preventing p62 accumulation in
autophagy-defective atg7.sup.-/- hepatocytes dramatically
suppresses the phenotype of steatohepatitis. In contrast,
neurodegeneration due to expression and accumulation of
polyglutamate expansion mutant proteins is greatly exacerbated by
allelic loss of beclin1 and defective autophagy. Thus, while not
intending to be bound by any theory of operation, autophagy is
believed to be involved in limiting the buildup of misfolded,
mutated proteins in p62-containing protein aggregates, which leads
to cellular deterioration and disease.
[0009] Analogous to a wound-healing response, chronic tumor cell
death in response to stress and induction of inflammation and
cytokine production may provide a non-cell-autonomous mechanism by
which tumorigenesis is promoted in autophagy-defective cells.
Autophagy-defective tumor cells also display an elevated DNA damage
response, gene amplification and chromosome instability in response
to stress, suggesting that autophagy limits genome damage as a
cell-autonomous mechanism of tumor suppression.
[0010] Therefore, while not intending to be bound by any theory of
operation, stimulating autophagy may be involved in limiting
disease progression, particularly neurodegeneration, liver disease,
and also cancer, by facilitating the elimination of protein
aggregates, damaged organelles, and the toxic consequences of their
accumulation.
[0011] Autophagy has been identified also as a survival pathway in
epithelial tumor cells that enables long-term survival to metabolic
stress. Tumor cells with defined defects in autophagy accumulate
p62-containing protein aggregates, DNA damage and die in response
to stress, whereas those with intact autophagy can survive for
weeks utilizing the autophagy survival pathway. Thus, autophagy
appears to be required to prevent tumor cell damage and to maintain
metabolism. Tumor cells can exploit this survival function to
remain dormant only to reemerge under more favorable conditions.
Interestingly, roughly half of human cancers may have impaired
autophagy, either due to constitutive activation of the PI-3 kinase
pathway or allelic loss of the essential autophagy gene beclin1,
rendering them particularly susceptible to metabolic stress and
autophagy inhibition.
[0012] Therefore, identification of the therapeutic means to
inhibit the autophagy survival pathway in tumor cells would be
advantageous. While not intending to be bound by any theory of
operation, this may be of value as many therapeutics currently in
use, such as kinase and angiogenesis inhibitors, inflict metabolic
stress, which increases the dependency on autophagy for survival.
Furthermore, tumor cells with impaired autophagy are particularly
vulnerable to metabolic stress and further therapeutic suppression
of autophagy may be able to exploit this vulnerability by promoting
cell death by metabolic catastrophe or the failure to mitigate cell
damage accumulation. Preclinical studies have been conducted using
hydroxychloroquine to inhibit lysosome acidification and thereby
autophagy in combination therapy. Specific inhibitors of the
autophagy survival pathway in tumor cells are may be of great value
in combination with agents such as angiogenesis and kinase
inhibitors that promote metabolic stress.
[0013] Thus, the autophagy pathway represents fertile ground for
novel therapeutic target identification for drug discovery for many
diseases for both acute treatment and also disease prevention.
[0014] Accordingly, a need exists to identify nucleic acid
sequences and their encoded proteins which are involved in
modulation of the autophagy pathway.
BRIEF SUMMARY OF THE INVENTION
[0015] In certain aspects, the present invention relates to methods
for identifying compounds that inhibit or stimulate the autophagy
pathway.
[0016] Further aspects relate to methods for identifying
individuals susceptible to or afflicted with a disease state
associate with an autophagy pathway defect.
[0017] Additional aspects relate to devices for detecting the
expression of autophagy-related genes.
[0018] Further aspects relate to kits for assaying expression of
autophagy-related genes.
[0019] Other aspects are readily apparent from the following
description.
BRIEF DESCRIPTION OF THE DRAWINGS
[0020] FIG. 1 illustrates a cell-based shRNA screen for rescue of
autophagy deficiency and p62 protein aggregate accumulation in
metabolic stress: Screen for autophagy stimulators.
[0021] FIG. 2 illustrates representative images of shRNAs that
modulate p62 aggregate elimination.
DETAILED DESCRIPTION OF THE INVENTION
[0022] The following definitions are provided to facilitate an
understanding of the present invention:
[0023] A "polynucleotide," "polynucleotide molecule" or
"polynucleotide sequence" refers to a chain of nucleotides. It may
refer to a DNA or RNA molecule, either single or double stranded
and, if single stranded, the molecule of its complementary sequence
in either linear or circular form. In some instances, the sequences
will be fully complementary (no mismatches) when aligned. In other
instances, there may be up to about a 30% mismatch in the
sequences.
[0024] The term "oligonucleotide," as used herein refers to
sequences, primers and probes, and is defined as a nucleic acid
molecule comprised of two or more ribo- or deoxyribonucleotides,
preferably more than three. The exact size of the oligonucleotide
will depend on various factors and on the particular application
and use of the oligonucleotide.
[0025] The term "probe" as used herein refers to either a probe for
a nucleic acid or a probe for a protein. When used in connection
with nucleic acids, a "probe" refers to an oligonucleotide,
polynucleotide or nucleic acid, either RNA or DNA, whether
occurring naturally as in a purified restriction enzyme digest or
produced synthetically, which is capable of annealing with or
specifically hybridizing to a nucleic acid with sequences
complementary to the probe. A probe may be either single stranded
or double stranded. The exact length of the probe will depend upon
many factors, including temperature, source of probe and method of
use. The probes herein are selected to be "substantially"
complementary to different strands of a particular target nucleic
acid sequence. This means that the probes must be sufficiently
complementary so as to be able to "specifically hybridize" or
anneal with their respective target strands under a set of
pre-determined conditions. Therefore, the probe sequence need not
reflect the exact complementary sequence of the target. For
example, a non complementary nucleotide fragment may be attached to
the 5' or 3' end of the probe, with the remainder of the probe
sequence being complementary to the target strand. Alternatively,
non complementary bases or longer sequences can be interspersed
into the probe, provided that the probe sequence has sufficient
complementarity with the sequence of the target nucleic acid to
anneal therewith specifically. When used in connection with a
polypeptide, a "probe" is a protein- or polypeptide-binding
substance or agent, capable of specifically binding a particular
protein or protein fragment to the substantial exclusion of other
proteins or protein fragments. Such binding agents may be any
molecule to which the protein or peptide specifically binds,
including DNA (for DNA binding proteins), antibodies (as described
in greater detail herein), cell membrane receptors, peptides,
cofactors, lectins, sugars, polysaccharides, cells, cell membranes,
organelles and organellar membranes.
[0026] "Array" refers to an ordered arrangement of at least two
probes on a substrate. At least one of the probes represents a
control or standard, and the other, a probe of diagnostic or
screening interest.
[0027] "Specific binding" refers to a special and precise
interaction between two molecules which is dependent upon their
structure, particularly their molecular side groups; for example,
the intercalation of a regulatory protein into the major groove of
a DNA molecule, the hydrogen bonding along the backbone between two
single stranded nucleic acids, or the binding between an epitope of
a protein and an agonist, antagonist, or antibody.
[0028] The term "specifically hybridize" refers to the association
between two single stranded nucleic acid molecules of sufficiently
complementary sequence to permit such hybridization under
predetermined conditions generally used in the art (sometimes
termed "substantially complementary"). For example, the term may
refer to hybridization of a nucleic acid probe with a substantially
complementary sequence contained within a single stranded DNA or
RNA molecule according to an aspect of the invention, to the
substantial exclusion of hybridization of the nucleic acid probe
with single stranded nucleic acids of non-complementary sequence.
When used in connection with the association between single
stranded nucleic acid molecules, the term "specifically bind" may
be used to indicate that the molecules "specifically hybridize" as
described herein.
[0029] An "antibody" or "antibody molecule" is any immunoglobulin,
including antibodies and fragments thereof, that binds to a
specific antigen. The term includes polyclonal, monoclonal,
chimeric, and bispecific antibodies. As used herein, antibody or
antibody molecule contemplates both an intact immunoglobulin
molecule and an immunologically active portion of an immunoglobulin
molecule such as those portions known in the art as Fab, Fab',
F(ab')2 and F(v).
[0030] "Sample" is used in its broadest sense as containing nucleic
acids, proteins, antibodies, and the like. A sample may comprise,
for example, a bodily fluid; the soluble fraction of a cell
preparation, or an aliquot of media in which cells were grown; a
chromosome, an organelle, or membrane isolated or extracted from a
cell; genomic DNA, RNA, or cDNA in solution or bound to a
substrate; a cell; a tissue or a tissue biopsy; a tissue print; a
fingerprint, buccal cells, skin, or hair; and the like. Bodily
fluids include, without limitation, whole blood, blood plasma,
blood serum, sputum, urine, sweat, and lymph.
[0031] As used herein, the term "subject" or "patient" refers to
both humans and animals, unless specified that the "subject" or
"patient" is an animal or a human. An "individual" also refers to
both humans and animals, unless specified that the "individual" is
an animal or a human. Animal subjects are preferably vertebrates,
and more preferably, mammals.
[0032] "Autophagy-associated" or "autophagy-related" as used herein
with respect to a disease, condition or disorder refers to that
which results from an increase or decrease in normal autophagy
function and/or that which may be treated and/or prevented by
modulation of the autophagy pathway. As used herein with respect to
a biological molecule, such as, for example, a polynucleotide or
polypeptide, "autophagy-associated" or "autophagy-related" refers
to a molecule for which alteration of the expression, abundance
and/or activity thereof leads to modulation of the autophagy
pathway.
[0033] In certain embodiments, the present invention relates to the
identification of genes whose expression modulates autophagy. These
genes and their gene products may represent targets for therapeutic
intervention in the autophagy pathway.
[0034] In accordance with aspects of the present invention, a
number of polynucleotides comprising at least a fragment of a gene
have been identified as representing molecules whose knockdown of
expression modulates the function of the autophagy pathway. In
certain aspects, knockdown of gene expression stimulates autophagy.
In other aspects, knockdown of gene expression inhibits
autophagy.
[0035] Embodiments of the invention include validation of the
candidate genes and gene fragments described herein using known
techniques for in vitro and in vivo analysis.
[0036] In accordance with various aspects of the present invention,
combinations, compositions, devices and kits are provided that may
be used in the practice of methods provided according to certain
embodiments of the invention.
[0037] Certain embodiments relate to methods of detection of
alterations in the autophagy pathway. Certain of these methods may
be used to detect conditions in which autophagy is reduced. Certain
of these methods may be used to detect conditions in which
autophagy is increased.
[0038] In accordance with certain aspects of the invention, a
combination is provided comprising a plurality of polynucleotide
molecules wherein the polynucleotide molecules encode gene products
associated with modulation of the autophagy pathway. In certain
embodiments, the combination comprises a plurality of
polynucleotides whose knockdown stimulates autophagy. In certain
embodiments, the plurality of polynucleotide molecules comprise two
or more molecules identified in Table 3 or fragments thereof.
[0039] In certain embodiments, the combination comprises a
plurality of polynucleotides whose knockdown inhibits autophagy. In
certain embodiments, the plurality of polynucleotide molecules
comprise two or more molecules identified in Table 4 or fragments
thereof.
[0040] An embodiment of the invention provides a method for
identifying compounds that modulate autophagy-associated gene
expression comprising: a) measuring standard expression by
measuring transcription or translation products of one or more of
the genes or gene fragments identified in Table 3 and/or 4, or
fragments thereof, in a standard sample in the absence of a test
compound; b) measuring test expression by measuring the
transcription or translation products of one or more of the genes
or gene fragments identified in Table 3 and/or 4, or fragments
thereof, in a test sample in the presence of the test compound; and
c) comparing the standard expression to the test expression,
wherein a change in the test expression compared to the standard
expression is indicative of an effect of the test compound on the
expression of genes whose expression modulates the autophagy
pathway. In certain embodiments, a plurality of two or more of the
genes or gene fragments identified in Table 3 and/or 4, or
fragments thereof are used.
[0041] One embodiment of the invention provides a method for
identifying compounds that inhibit or stimulate the autophagy
pathway for treatment of a disease state associated with an
autophagy pathway defect, comprising measuring the effect of one or
more test compounds on the inhibition or stimulation of a product
of one or more of the genes or gene fragments identified in Table 3
or Table 4.
[0042] An embodiment provides a method for the detection of
differential expression of autophagy-associated polypeptides in a
sample, comprising the steps of: a) reacting protein binding
molecules with polypeptides of the sample, thereby allowing
specific binding to occur, wherein the polypeptides bound by the
protein-binding molecules comprise one or more polypeptides encoded
by the genes or gene fragments identified in Table 3 and/or Table 4
or fragments thereof; b) detecting specific binding; and c)
comparing the specific binding in the sample with that of a
standard, wherein differences between the standard and sample
specific binding indicate differential expression of polypeptides
in the sample. In certain embodiments, the protein-binding
molecules are directed to polypeptides comprising a plurality of
two or more polypeptides encoded by the genes or gene fragments
identified in Table 3 and/or Table 4 or fragments thereof.
[0043] Another embodiment provides a method for the detection of
differential expression of autophagy-associated nucleic acids in a
sample, comprising the steps of: a) hybridizing polynucleotides
comprising one or more molecules identified in Table 3 and/or Table
4 or fragments thereof with nucleic acids of the sample, thereby
forming one or more hybridization complexes; b) detecting the
hybridization complexes; and c) comparing the hybridization
complexes with those of a standard, wherein differences between the
standard and sample hybridization complexes indicate differential
expression of nucleic acids in the sample. In certain embodiments,
the polynucleotides comprise a plurality of two or more molecules
identified in Table 3 and/or Table 4 or fragments thereof.
[0044] Another embodiment comprises a composition of matter
comprising one or more probes for detecting expression of
autophagy-associated genes, wherein the probes comprise one or more
of: a) nucleic acid molecules that specifically hybridize to one or
more of the genes or gene fragments identified in Table 3 and/or
Table 4, or fragments thereof; or b) polypeptide binding agents
that specifically bind to polypeptides produced by expression of
one or more nucleic acid molecules comprising sequences selected
from one or more of genes or gene fragments identified in Table 3
and/or Table 4, or fragments thereof. In certain embodiments, the
composition of matter comprises a collection of two or more
probes.
[0045] Another embodiment provides a device for detecting
expression of a plurality of autophagy-related genes, comprising a
substrate to which is affixed, at known locations, a plurality of
probes, wherein the probes comprise: a) a plurality of
oligonucleotides or polynucleotides, each of which specifically
hybridizes to a different sequence selected from any of the
sequences identified in Table 3 and/or Table 4 or fragments
thereof; or b) a plurality of polypeptide binding agents, each of
which specifically binds to a different polypeptide or fragment
thereof produced by expression of a nucleic acid molecule
comprising a sequence selected from the genes or gene fragments
comprising any of the sequences identified in Table 3 and/or Table
4 or fragments thereof.
[0046] In certain embodiments, a device is provided for detecting
the expression of a plurality of autophagy-related genes associated
with an autophagy pathway defect, said device comprising a
substrate to which is affixed at known locations a plurality of
probes, wherein the probes comprise: [0047] a) a plurality of
oligonucleotides or polynucleotides, each of which specifically
binds to a different sequence selected from any of the sequences
identified in Table 3 or Table 4 or fragments thereof; or [0048] b)
a plurality of polypeptide binding agents, each of which
specifically binds to a different polypeptide or fragment thereof
produced by expression of a gene or gene fragment comprising any of
the sequences identified in Table 3 or Table 4 or fragments
thereof.
[0049] Another embodiment provides a method for measuring the
effect of a test compound on expression of an autophagy-associated
gene, wherein the gene is selected from the group consisting of the
genes or gene fragments identified in Table 3 and/or 4, the method
comprising measuring production of transcription or translation
products produced by expression of the gene or gene fragment in the
presence or absence of the test compound, wherein a change in the
production of transcription or translation products in the presence
of the test compound is indicative of an effect of the test
compound on expression of the gene or gene fragment.
[0050] In an embodiment, the gene expression is measured by
providing a DNA construct comprising a reporter gene coding
sequence operably linked to transcription regulatory sequences of
the autophagy-associated gene, and measuring formation of a
reporter gene product in the presence or absence of the test
compound.
[0051] Another embodiment provides a kit for assaying the
expression of autophagy-related genes, comprising at least one
container comprising a collection of two or more probes, wherein
the probes comprise: a) oligonucleotides or polynucleotides that
specifically hybridize to two or more genes or gene fragments
comprising any of the sequences identified in Table 3 and/or Table
4, or fragments thereof; or b) polypeptide binding agents that
specifically bind to polypeptides produced by expression of two or
more genes or gene fragments comprising any of the sequences
identified in Table 3 and/or Table 4, or fragments thereof. The kit
preferably comprises instructions for performing an assay of gene
expression.
[0052] In certain embodiments, the invention provides a kit for
assaying the expression of autophagy-related genes associated with
an autophagy pathway defect, comprising at least one container and
a collection of two or more probes, wherein the probes comprise:
[0053] a) oligonucleotides or polynucleotides that specifically
bind to two or more genes or gene fragments comprising any of the
sequences identified in Table 3 or Table 4, or fragments thereof;
or [0054] b) polypeptide binding agents that specifically bind to
polypeptides produced by expression of two or more genes or gene
fragments comprising any of the sequences identified in Table 3 or
Table 4, or fragments thereof. The kit preferably comprises
instructions for performing an assay of gene expression.
[0055] The invention provides, in certain embodiments, methods for
identifying compounds that are useful in modulating the autophagy
pathway. Preferably, the methods include contacting at least one
polypeptide encoded by the genes and/or gene fragments identified
in Table 3 and/or Table 4 with a test substance and determining
whether the test substance binds to the polypeptide. Further, in
certain embodiments of the invention, a test substance may be
determined to stimulate or inhibit the biological activity of the
relevant gene product comprising at least one polypeptide encoded
by the genes and/or gene fragments identified in Table 3 and/or
Table 4 and thereby be identified as a compound useful for the
modulation of the autophagy pathway. Such assays may, in certain
embodiments, be performed in vitro and may, in certain embodiments,
be performed in a cell-based assay. In some embodiments, substances
identified as modulating expression or biological activity in vitro
may be further tested in vivo to confirm relevant and effective
activity.
[0056] Test substances or compounds contemplated by aspects of the
invention include compounds from chemical libraries, including
natural products and/or synthetic products from combinatorial
chemical synthesis. Such substances may include, without
limitation, polypeptides, oligonucleotides, polynucleotides, or
organic molecules.
[0057] In a further embodiment is provided a method of modulating
autophagy-associated gene expression in a cell by administering an
effective amount of a composition under appropriate conditions to
affect the expression of at least one gene associated with
autophagy having a sequence selected from the sequences identified
in Table 3 and/or Table 4, or fragments thereof.
[0058] In preferred embodiments, the composition comprises an
inhibitor of gene expression. The inhibitor of gene expression may
be selected from molecules including, but not limited to, an
antisense RNA, a morpholino polynucleotide, and an interfering RNA
(RNAi).
[0059] According to a still further aspect of the invention, there
is provided a genetically-modified non-human animal that has been
transformed to express higher, lower or absent levels of a protein
according to any one of the aspects of the invention described
herein. Preferably, said genetically-modified animal is a
transgenic or knockout animal. Preferably, the genetically-modified
animal is a rodent, most preferably a mouse.
[0060] An embodiment of the invention also provides a method for
screening for a substance effective to treat an
autophagy-associated disease condition, by contacting a non-human
genetically-modified animal as described above with a candidate
substance and determining the effect of the substance on the
physiological state of the animal.
[0061] Certain embodiments of the invention provide methods and
kits for diagnosis of, determining susceptibility to and/or
developing a prognosis for an autophagy-associated disease state in
a subject. In certain aspects, these may involve tests on subject
samples. In certain embodiments, these may be nucleic acid based
tests or polypeptide-based tests. In some embodiments, the method
or kit may include probes that bind to at least one polynucleotide
encoding an autophagy-associated polypeptode. In some embodiments,
the a plurality of two or more probes may be used. In some
embodiments, the method or kit may include polypeptide binding
agents that bind to at least one autophagy-associated polypeptide.
In some embodiments, a plurality of two or more polypeptide binding
agents may be used. In certain embodiments, the polypeptide-binding
agent comprises antibodies and/or antigen-binding portions of an
antibody that specifically binds to one or more
autophagy-associated polypeptides. Preferably, the
autophagy-associated polypeptides are encoded by the gene or gene
fragments identified in Table 3 and/or Table 4.
[0062] One embodiment provides a method for identifying individuals
susceptible to or afflicted with a disease state associated with an
autophagy pathway defect, comprising testing a biological sample
from an individual for a characteristic of one or more polypeptides
produced by expression of one or more of the genes or gene
fragments identified in Table 3 or Table 4 that is indicative of
said disease state, wherein said characteristic is selected from
the presence of at least one of said polypeptides, the absence of
at least one of said polypeptides, an elevated level of at least
one of said polypeptides, a reduced level of at least one of said
polypeptides and, for two or more of said polypeptides,
combinations thereof.
[0063] An embodiment provides a method to diagnose or develop a
prognosis for an autophagy-related disease in a subject, the method
comprising: a) obtaining a sample from the subject; b) measuring in
the sample the production of transcription or translation products
produced by the expression of one or more autophagy-associated
genes or gene fragments comprising any of the sequences identified
in Table 3 and/or Table 4, or fragments thereof; c) comparing the
transcription or translation products of the sample with that of a
standard, wherein a difference in the expression of any of the
autophagy-associated genes or gene fragments is indicative of
autophagy-related disease.
[0064] In one embodiment is provided a kit for the diagnosis of an
autophagy-associated disease in a subject comprising polynucleotide
probes that specifically bind to one or more autophagy-associated
polynucleotides or a fragment thereof. Preferably the
autophagy-associated polynucleotides or fragments thereof are
selected from the polynucleotide sequences identified in Table 3
and/or Table 4 or fragments thereof. In certain embodiments, the
kit comprises a plurality of two or more polynucleotide probes that
specifically bind to polynucleotide sequences identified in Table 3
and/or Table 4 or fragments thereof. Preferably, the kit comprises
also instructions for use.
[0065] The invention also provides kits for diagnosis of
autophagy-associated conditions from patient samples that may be
nucleic acid based tests or polypeptide-based tests. In some
embodiments, the kit contains at least one polynucleotide that
binds to a polynucleotide encoding an autophagy-related gene
product. In some embodiments, the kit contains, preferably in
separate containers, a plurality of probes to detect two or more
polynucleotides encoding one or more autophagy-associated gene
products. In preferred embodiments, the gene products are encoded
by one or more of the genes or gene fragments identified in Table 3
and/or Table 4. In other embodiments, the kit contains at least one
polypeptide binding agent that specifically binds to at least one
autophagy-associated polypeptide. In some embodiments, the kit
contains, preferably in separate containers, a plurality of
polypeptide binding agents (or mixtures thereof) to detect one or
more autophagy-associated polypeptide. In certain embodiments, the
polypeptide binding agent may be an antibody or antigen-binding
portion of an antibody. In certain embodiments, the
autophagy-associated polypeptides include at least one polypeptide
encoded by the genes or gene fragments identified in Table 3 and/or
Table 4. In certain embodiments, the autophagy-associated
polypeptides identified by the kit include a plurality of two or
more polypeptides encoded by the genes or gene fragments identified
in Table 3 and/or Table 4. In certain embodiments, the kits may
also include instructions for use.
[0066] In certain embodiments, methods according to the invention
may be used for high-throughput screening assays.
[0067] In certain embodiments, methods and kits useful in the
methods of the invention may utilize nucleic acid, antibody and/or
polypeptide arrays.
[0068] Using a cell-based loss-of function screen, the present
inventors have identified candidate genes whose expression is
involved in the autophagy pathway. In particular, the screen has
been used to identify genes whose knockdown stimulates autophagy.
Results from this screen are shown in Table 1. The screen has also
been used to identify genes whose knockdown inhibits autophagy.
Results from this screen are shown in Table 2.
[0069] A high-efficiency delivery method that enables stable
long-term gene suppression in a broad range of cell types is
virus-mediated integration of an RNAi expression cassette. After
integration, the cassette produces a short dsRNA molecule, usually
in the form of a hairpin structure, a short or small hairpin RNA
(shRNA), which is processed into active small interfering RNA
(siRNA). Although many types of viruses are suitable for this
purpose, lentiviral vectors generate viruses of both high titer and
broad tropism, permitting the infection of both dividing and
nondividing cells. Lentiviral shRNA libraries for mouse gene clones
were utilized that allow gene silencing in most dividing and
nondividing cell types.
[0070] An image based, arrayed shRNA screen was employed.
Lentiviral shRNA libraries developed by the RNA Consortium (TRC) at
the Broad Institute were used in a cell-based screen. The screens
utilized the publicly available kinase and vesicle trafficking
lentiviral library subsets at the Broad Institute, as well as a
custom library containing shRNAs targeting mouse GTPases.
Lentiviruses are high-titer, individual clones with representation
of at least five independent hairpins for each target gene supplied
in a high-throughput format (Root, D. E., et al. (2006), Nature
Methods 3, 715-719.) Fluorescence image analysis was used to
capture the data. Gene were identified that were shown to promote
or suppress autophagy (bimodal analysis).
[0071] The high content arrayed shRNA screen used to identify
autophagy modulators utilized autophagy defective beclin1.sup.+/-
iBMK cells stably expressing the autophagy substrate EGFP-p62
(Mathew, R., Karantza-Wadsworth, V., and White, E. (2009) Methods
Enzymol 453, 53-81; Mathew, R., et al. (2009) Cell 137, 1062-1075;
and Mathew, R., et al. (2007) Genes Dev 21, 1367-1381). p62
accumulates and aggregates in response to metabolic stress and
requires autophagy for degradation. p62 also accumulates in
degenerative neuronal and liver diseases and in autophagy-defective
mouse tissues, beclin1.sup.+/- and atg5.sup.-/- iBMK cells, and
tumors. Genes were identified whose inactivation compensates for
defective autophagy and restores p62 protein turnover. Since the
image analysis captured every hairpin's p62 aggregation score
(EGFP-p62 intensity divided by the nuclei in the field) it was also
possible to identify genes whose inactivation lead to further
accumulation of p62 aggregates, predicted to be autophagy
inhibitors (FIG. 2). This was possible because the cell line
employed in this screen is autophagy impaired rather than fully
autophagy defective. Therefore, the disposition of p62 in the test
cells serves as readout for both autophagy promotion (p62
degradation, low p62) and inhibition (autophagy inhibition, high
p62) and is the basis for the identification of autophagy
modulators in cell-based screens.
[0072] The shRNA libraries were screened using autophagy-impaired
test cells expressing a marker of protein aggregation, subjecting
the test cell to metabolic stress, and performing analysis on the
test cell to determine the level of the marker. The marker of
protein aggregation is a p62 protein linked to enhanced green
fluorescent protein (EGFP) label. Image analysis is performed to
determine the level of p62 aggregates in cells. The level of the
marker found in p62 aggregates in the test cell is compared with
that of a control cell. A lower level of p62 aggregates comprising
the marker in the test cell compared to that demonstrated by the
control cell demonstrates the rescue of the impairment in p62
clearance, indicating the lowered expression of a gene whose
knockdown stimulates autophagy. A greater level of p62 aggregates
in a test cell compared to that of a control cell demonstrates
suppression of p62 clearance, indicating the knockdown of a gene
whose lowered level of expression leads to inhibition of
autophagy.
[0073] The cell-based screen utilized autophagy-deficient
beclin1.sup.+/- immortalized baby mouse kidney (iBMK) cells stably
expressing EGFP-p62. p62 accumulates and aggregates in response to
metabolic stress and requires autophagy for degradation. FIG. 1
illustrates a cell-based shRNA screen for rescue of autophagy
deficiency and p62 protein aggregate accumulation in metabolic
stress and therefore represents a screen for autophagy stimulators.
The autophagy-deficient beclin1.sup.+/- iBMK cell line stably
expressing EGFP-p62 accumulates p62-containing protein aggregates
under stress, which fail to be cleared following recovery. Those
shRNAs that facilitate p62 aggregate clearance, compensating for
defective autophagy, are identified. The autophagy wild type
beclin1.sup.+/+ iBMK cell line stably expressing EGFP-p62 that
effectively clears p62 aggregates following stress is used as a
positive control.
[0074] FIG. 2 illustrates representative images of cells contacted
with shRNAs that modulate p62 aggregate elimination. The screen is
designed to identify genes whose loss results in restoration of
autophagy (autophagy stimulators), manifested by successful
clearance of p62 aggregates following a time course of stress and
recovery. mTOR, a master negative regulator of autophagy, is shown
here as an example of a gene whose loss restores autophagy and
clearance of p62. Alternatively, loss of some genes is predicted to
further inhibit autophagy (autophagy inhibitors). Loss of Ikbkb, a
known autophagy promoter (Criollo, A, et al. EMBO J 29, 619-631),
results in marked accumulation of p62. This accumulation is greater
than observed in cells infected with an shRNA targeting
luciferase.
[0075] The shRNAs shown to promote p62 elimination (autophagy
stimulators) identify potential targets for drug discovery efforts
for development of modulators of autophagy, including autophagy
inhibitors. While not intending to be bound by any theory of
operation, autophagy inhibitors are potentially useful as
anti-cancer therapeutics by promoting cancer cell death.
[0076] The shRNAs shown to enhance p62 accumulation (autophagy
inhibitors) identify potential targets for drug discovery efforts
for development of modulators of autophagy, including autophagy
stimulators. While not intending to be bound by any theory of
operation, autophagy stimulators are potentially useful in
preventing or delaying disease manifestation in the setting of
cancer, neurodegenerative conditions, Crohn's disease, liver
disease, aging and inflammatory diseases and in combating
infections.
EXAMPLES
[0077] The following examples serve to more fully describe the
manner of using the above-described invention. It is understood
that these examples in no way serve to limit the true scope of this
invention, but rather are presented for illustrative purposes.
Materials and Methods
Cell Line
[0078] Murine kidney epithelial cells were isolated from
beclin1.sup.+/- mice and immortalized with dominant negative p53
and EIA as described previously (Degenhardt, K., and White, E.
(2006). Clin Cancer Res 12, 5298-5304; Mathew, R., Degenhardt, K.,
Haramaty, L., Karp, C. M., and White, E. (2008) Methods Enzymol
446, 77-106.). The cells were subsequently engineered to
overexpress Bcl-2 and eGFP-p62. Thus, these cells, known as 3BC2
EGFP-P62, contain an autophagy defect, are apoptotically impaired,
and stably express EGFP-P62. (Mathew, R., et al. (2009). Cell 137,
1062-1075.).
Screening Protocol for Beclin.sup.+/- eGFP-P62
[0079] Cells were plated into black barcoded 384 well plates
(Corning 8793BC) at a density of 700 cells/well by the Biotek
microfill and allowed to attach overnight. Infection and media
changes for plates were achieved by use of two robotic liquid
handlers at the Broad Institute, the Perkin Elmer Janus and EP3.
Each viral plate was used to infect four target plates. Each virus
plate contained 20 control hairpins (targeting either RFP,
luciferase, or EGFP) in addition to wells containing no virus. Two
hairpins targeting p62 were spiked into each plate at the time of
infection to ensure that positive and negative controls were
present on all plates. Immediately prior to infection, media was
changed with the Janus robot (Perkin Elmer) to DMEM containing 8
ug/mL polybrene. The Perkin Elmer EP3 robot was used to add 6 ul of
virus to each well. Cells were spin infected (2250 rpm 30 mins,
30.degree. C.) in the presence of 6 ul of virus and Bug/ml of
polybrene before returning to the 37 C incubator. Virus and
polybrene containing media was removed 4 hours post infection and
cells were incubated in normal growth media overnight (DMEM high
glucose, 10% FBS, 1% penicillin streptomycin (PS)). Twenty four
hours post infection, media was changed with the Janus to DMEM
containing 3 ug/mL puromycin (for the three puro plus plates) or
DMEM alone (for the puro minus replicate). Selection was allowed to
continue for 72 hrs.
[0080] The assay employed in this screen is predicated on the
ability of autophagy competent cells to successfully eliminate p62
aggregates that accumulate during metabolic stress during a
recovery phase during which time oxygen and glucose are restored.
Optimization experiments were conducted comparing the ability of
autophagy competent beclin1.sup.+/+-EGFP-p62 cells (WB3-EGFP-p62)
and autophagy deficient beclin1.sup.+/--EGFP-p62 cells
(3Bc2-EGFP-p62) to eliminate p62 aggregates during various time
courses of metabolic stress (1% oxygen, glucose deprivation) and
recovery within the setting of 384 well plates post infection and
selection with puromycin. 7.5 hours of metabolic stress followed by
18 hours of recovery in high glucose DMEM 10% FBS was optimal, and
these conditions were chosen for the large-scale screen.
[0081] Following puromycin selection, media containing DMEM high
glucose was removed, and cells were washed twice in ischemia media
(DMEM containing no glucose, 10% FBS, 1% PS) to remove residual
glucose in wells prior to transfer into a hypoxia incubator set to
1% oxygen for 7.5 hours. They were then transferred to an incubator
which could lower ambient oxygen levels to 1% by virtue of its
attachment to a nitrogen tank. Cells stayed in this 1% oxygen, no
glucose conditions, referred to as metabolic stress, for 7.5 h.
[0082] At the conclusion of the metabolic stress, normal growth
media (DMEM high glucose 10% FBS, 1% PS) was added to the plates
and the cells were allowed to recover overnight at 37.degree. C. 18
hours post recovery, media was removed from plates, and cells were
fixed by addition of 4% paraformaldehyde/PBS for 10 mins at RT.
[0083] Nuclei were visualized by inclusion of Hoechst 33342 at a
dilution of 1:10,000 in the fixation solution. Plates were washed
3.times. with the ELx405 automated plate washer (Biotek). 80 ul of
filtered PBS was left in each well at the end of washing to allow
for evaporation during imaging.
[0084] Plates were imaged on the Arrayscan VTI (Thermo Scientific)
housed within the Genome technology Core of the Whitehead Institute
using a modified version of the Cellomics compartmental analysis
bioapplication. Nuclei were visualized in channel 1. EGFP-p62
aggregates were visualized in channel 2. Nine images per channel
were captured for each field, with an autofocus field interval of
3. MEAN_valid object count channel 1 represents the mean nuclear
count within the field. MEAN_ring spot average integrated intensity
channel 2 represents the mean intensity of the p62 aggregates in
the field. To properly identify the p62 aggregates the following
settings were employed: Spot kernel radius: 10, ring distance from
nucleus: 0, ring width: 10 pixels. Data was exported to Excel for
further analysis. Data quality (batch-to-batch variation,
similarity of replicates) was examined with Spotfire decision
software and RNAeyes, in house software developed by The RNAi
Consortium (TRC) of the Broad Institute.
[0085] A p62 aggregate score equal to Mean Ring Spot total
intensity/Mean nuclei was calculated for each well. Viral
infections were done in quadruplicate, with three plates receiving
puromycin, one not. A comparison of the nuclei counts from the
puro+/puro- plates allowed calculation of the infection efficiency
of each hairpin. Hairpins with less than 1500 nuclei per well or
those that had an infection efficiency less than 25% were omitted
from subsequent analysis. A robust Z-score, a standard metric for
high throughput assays (Birmingham A., et al. (2009) Nat Methods 6,
569-575), was calculated for each well. The three puromycin
selected replicates were averaged, and this value was used for
further analysis.
[0086] The in-house Gene-E software ranked genes at both a hairpin
and a gene level. Attached to this application are candidate
results (`hits`) from either end of our analysis: those that
resulted in profound elimination of p62, predicted to be autophagy
inducers (Table 1), and those that resulted in profound
accumulation of p62, predicted to be autophagy inhibitors (Table
2). These tables represent the weighted sum ranking of the data. In
this metric, 75% of the score is based on the robust z-score of the
second best hairpin for a given gene, while the other 25% of the
score is based on the rank of the robust z-score of the best
hairpin. Similar data was obtained when three other analysis
measures were employed: cut-off based on a given standard deviation
from controls, second best ranking, or RNA Interference Gene
Enrichment Rank (RIGER) analysis based on the KS statistic as
described previously (Luo B., et al. (2008) Proc Natl Acad Sci USA
105, 20380-20385).
[0087] A subset of viral plates were re-screened to ensure
reproducibility of the assay and analyses.
Example 1
[0088] Table 1 shows results of a screen that led to elimination of
p62.
TABLE-US-00001 TABLE 1 Symbol GeneID Gene Rank Mtor 56717 1 Tssk3
58864 2 Pik3c3 225326 3 Cask 12361 4 Lrguk 74354 5 GeneID: 218456
218456 6 Rab9 56382 7 GeneID: 381390 381390 8 Gm4922 237300 9 Cdk8
264064 10 Ephb1 270190 11 Prkaca 18747 12 Kpna2 16647 13 Pldn 18457
14 Scfd1 76983 15 Ripk3 56532 16 Trib3 228775 17 Vapa 30960 18
Trrap 100683 19 Mpp3 13384 20 GeneID: 381082 381082 21 Mapk14 26416
22 Adk 11534 23 Ern1 78943 24 Hip1r 29816 25 Nek5 330721 26 Alpk3
116904 27 4932415M13Rik 211496 28 Vps33b 233405 29 1810024B03Rik
329509 30 Chmp1a 234852 31 Atp6v0a1 11975 32 Ddr2 18214 33 Cdk6
12571 34 Stxbp3a 20912 35 Map4k3 225028 36 Egfr 13649 37 Tpr 108989
38 Tlk2 24086 39 Rhoh 74734 40 Sar1a 20224 41 Vta1 66201 42 Rab34
19376 43 Brdt 114642 44 GFP -10 45 GeneID: 384481 384481 46 Dgka
13139 47 Rabl2a 68708 48 Snx10 71982 49 Rhoa 11848 50 Map3k11 26403
51 Gm5374 385049 52 D1g4 13385 53 Rab7l1 226422 54 Vamp8 22320 55
N4bp2 333789 56 Arf3 11842 57 GeneID: 381309 381309 58 Plk2 20620
59 Cpne3 70568 60 Hip1 215114 61 Musk 18198 62 Rab39b 67790 63 Akt1
11651 64 Arhgap24 231532 65 Eif2ak1 15467 66 Pfkfb4 270198 67
Txndc3 73412 68 Pim1 18712 69 5730410E15Rik 319613 70 1190002A17Rik
68870 71 Gvin1 74558 72 Pank4 269614 73 Bmpr1a 12166 74 Grk4 14772
75 Pip5k1a 18720 76 Prkcz 18762 77 Nek3 23954 78 Pgk1 18655 79 Kras
16653 80 Tssk6 83984 81 Rab2a 59021 82 Mertk 17289 83 Ap1m2 11768
84 Snx2 67804 85 Ilk 16202 86 Dgkk 331374 87 Csnk1d 104318 88
Rps6kb2 58988 89 Map3k4 26407 90 Ca1m1 12313 91 Trim24 21848 92 Fyn
14360 93 Sh3bp5 24056 94 Fn3k 63828 95 Ippk 75678 96 Tspan1 66805
97 Cd81 12520 98 Met 17295 99 Pdgfra 18595 100 Vrk2 69922 101 Gem
14579 102 Camkk1 55984 103 Pfkl 18641 104 Stx8 55943 105 Tspan9
109246 106 Stx17 67727 107 Tm4sf1 17112 108 Chmp4c 66371 109 Tbk1
56480 110 Dgkh 380921 111 Ephb4 13846 112 Rhof 23912 113 Pik3cd
18707 114 Mark1 226778 115 Tspan2 70747 116 Dync1li1 235661 117
Trim27 19720 118 Aurkc 20871 119 Itpkb 320404 120 Cav1 12389 121
Bub1 12235 122 Rap1b 215449 123 Mapk10 26414 124 Mapk8 26419 125
Rab24 19336 126 Kdr 16542 127 Rab2b 76338 128 Irak3 73914 129
Map3k8 26410 130 Csnk1a1 93687 131 Rhobtb1 69288 132 Mast2 17776
133 Raf1 110157 134 Arl11 219144 135 Dgkq 110524 136 Arfrp1 76688
137 Mknk2 17347 138 Erbb3 13867 139 Rheb 19744 140 Clk4 12750 141
Map2k1 26395 142 Sbk1 104175 143 Clk3 102414 144 Irak1 16179 145
Pik3cb 74769 146 Map3k12 26404 147 Tk1 21877 148 Aatk 11302 149
Fastkd2 75619 150
TABLE-US-00002 TABLE 3 GeneID RefSeq Species Description 270198
NM_001039217 MUS 6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE- MUSCULUS
2,6-BIPHOSPHATASE 4 270198 NM_001039215 MUS
6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE- MUSCULUS 2,6-BIPHOSPHATASE 4
270198 NM_001039216 MUS 6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE- MUSCULUS
2,6-BIPHOSPHATASE 4 270198 NM_173019 MUS
6-PHOSPHOFRUCTO-2-KINASE/FRUCTOSE- MUSCULUS 2,6-BIPHOSPHATASE 4
11768 NM_009678 MUS ADAPTOR PROTEIN COMPLEX AP-1, MU 2 MUSCULUS
SUBUNIT 11534 NM_134079 MUS ADENOSINE KINASE MUSCULUS 11842
NM_007478 MUS ADP-RIBOSYLATION FACTOR 3 MUSCULUS 76688 NM_029702
MUS ADP-RIBOSYLATION FACTOR RELATED MUSCULUS PROTEIN 1 219144
NM_177337 MUS ADP-RIBOSYLATION FACTOR-LIKE 11 MUSCULUS 116904
NM_054085 MUS ALPHA-KINASE 3 MUSCULUS 11302 NM_007377 MUS
APOPTOSIS-ASSOCIATED TYROSINE MUSCULUS KINASE 11975 NM_016920 MUS
ATPASE, H+ TRANSPORTING, LYSOSOMAL MUSCULUS V0 SUBUNIT A1 20871
NM_020572 MUS AURORA KINASE C MUSCULUS 333789 NM_001024917 MUS BCL3
BINDING PROTEIN MUSCULUS 12166 NM_009758 MUS BONE MORPHOGENETIC
PROTEIN MUSCULUS RECEPTOR, TYPE 1A 114642 NM_054054 MUS
BROMODOMAIN, TESTIS-SPECIFIC MUSCULUS 12235 NM_009772 MUS BUDDING
UNINHIBITED BY MUSCULUS BENZIMIDAZOLES 1 HOMOLOG (S. CEREVISIAE)
17289 NM_008587 MUS C-MER PROTO-ONCOGENE TYROSINE MUSCULUS KINASE
55984 NM_018883 MUS CALCIUM/CALMODULIN-DEPENDENT MUSCULUS PROTEIN
KINASE KINASE 1, ALPHA 12361 NM_009806 MUS
CALCIUM/CALMODULIN-DEPENDENT MUSCULUS SERINE PROTEIN KINASE (MAGUK
FAMILY) 12313 NM_007589 MUS CALMODULIN 1 MUSCULUS 12313 NM_007590
MUS CALMODULIN 1 MUSCULUS 12313 NM_009790 MUS CALMODULIN 1 MUSCULUS
104318 NM_027874 MUS CASEIN KINASE 1, DELTA MUSCULUS 104318
NM_139059 MUS CASEIN KINASE 1, DELTA MUSCULUS 93687 NM_146087 MUS
CASEIN KINASE I-ALPHA MUSCULUS 12389 NM_007616 MUS CAVEOLIN,
CAVEOLAE PROTEIN 1 MUSCULUS 12520 NM_133655 MUS SP. CD 81 ANTIGEN
12520 NM_133655 MUS CD 81 ANTIGEN MUSCULUS 12750 NM_007714 MUS CDC
LIKE KINASE 4 MUSCULUS 102414 NM_007713 MUS CDC-LIKE KINASE 3
MUSCULUS 70568 NM_027769 MUS COPINE III MUSCULUS 12571 NM_009873
MUS CYCLIN-DEPENDENT KINASE 6 MUSCULUS 264064 NM_181570 MUS
CYCLIN-DEPENDENT KINASE 8 MUSCULUS 264064 NM_153599 MUS
CYCLIN-DEPENDENT KINASE 8 MUSCULUS 331374 NM_177914 MUS
DIACYLGLYCEROL KINASE KAPPA MUSCULUS 13139 NM_016811 MUS
DIACYLGLYCEROL KINASE, ALPHA MUSCULUS 380921 NM_001081336 MUS
DIACYLGLYCEROL KINASE, ETA XM_895030 MUSCULUS 380921 XM_902438 MUS
DIACYLGLYCEROL KINASE, ETA MUSCULUS 380921 XM_484397 MUS
DIACYLGLYCEROL KINASE, ETA MUSCULUS 380921 XM_916777 MUS
DIACYLGLYCEROL KINASE, ETA MUSCULUS 380921 XM_924467 MUS
DIACYLGLYCEROL KINASE, ETA MUSCULUS 110524 NM_199011 MUS
DIACYLGLYCEROL KINASE, THETA MUSCULUS 18214 NM_022563 MUS DISCOIDIN
DOMAIN RECEPTOR FAMILY, MUSCULUS MEMBER 2 13385 NM_007864 MUS
DISCS, LARGE HOMOLOG 4 (DROSOPHILA) MUSCULUS 235661 NM_146229 MUS
DYNEIN CYTOPLASMIC 1 LIGHT MUSCULUS INTERMEDIATE CHAIN 1 78943
NM_023913 MUS ENDOPLASMIC RETICULUM (ER) TO MUSCULUS NUCLEUS
SIGNALLING 1 270190 NM_173447 MUS EPH RECEPTOR B1 MUSCULUS 13846
NM_010144 MUS EPH RECEPTOR B4 MUSCULUS 13649 NM_007912 MUS
EPIDERMAL GROWTH FACTOR RECEPTOR MUSCULUS 13649 NM_207655 MUS
EPIDERMAL GROWTH FACTOR RECEPTOR MUSCULUS 15467 NM_013557 MUS
EUKARYOTIC TRANSLATION INITIATION MUSCULUS FACTOR 2 ALPHA KINASE 1
56717 NM_020009 MUS FK506 BINDING PROTEIN 12-RAPAMYCIN MUSCULUS
ASSOCIATED PROTEIN 1 56717 NM_001039554 MUS FK506 BINDING PROTEIN
12-RAPAMYCIN MUSCULUS ASSOCIATED PROTEIN 1 63828 NM_001038699 MUS
FRUCTOSAMINE 3 KINASE MUSCULUS 63828 NM_022014 MUS FRUCTOSAMINE 3
KINASE MUSCULUS 14360 NM_008054 MUS FYN PROTO-ONCOGENE MUSCULUS
14772 NM_019497 MUS G PROTEIN-COUPLED RECEPTOR KINASE MUSCULUS 2,
GROUCHO GENE RELATED (DROSOPHILA) 14579 NM_010276 MUS GTP BINDING
PROTEIN (GENE MUSCULUS OVEREXPRESSED IN SKELETAL MUSCLE) 74558
NM_001039160 MUS GTPASE, VERY LARGE INTERFERON MUSCULUS INDUCIBLE 1
74558 NM_029000 MUS GTPASE, VERY LARGE INTERFERON MUSCULUS
INDUCIBLE 1 29816 NM_145070 MUS HUNTINGTIN INTERACTING PROTEIN 1
MUSCULUS RELATED 237300 NM_177706 MUS HYPOTHETICAL PROTEIN
4933423E17 MUSCULUS 228775 NM_175093 MUS INDUCED IN FATTY LIVER
DYSTROPHY 2 MUSCULUS 228775 NM_144554 MUS INDUCED IN FATTY LIVER
DYSTROPHY 2 MUSCULUS 75678 NM_199056 MUS INOSITOL
1,3,4,5,6-PENTAKISPHOSPHATE MUSCULUS 2-KINASE 320404 NM_001081175
MUS INOSITOL 1,4,5-TRISPHOSPHATE 3-KINASE B XM_205854 MUSCULUS
320404 XM_923874 MUS INOSITOL 1,4,5-TRISPHOSPHATE 3-KINASE B
MUSCULUS 320404 XM_915655 MUS INOSITOL 1,4,5-TRISPHOSPHATE 3-KINASE
B MUSCULUS 320404 XM_900404 MUS INOSITOL 1,4,5-TRISPHOSPHATE
3-KINASE B MUSCULUS 16202 NM_010562 MUS INTEGRIN LINKED KINASE
MUSCULUS 16179 NM_008363 MUS INTERLEUKIN-1 RECEPTOR-ASSOCIATED
MUSCULUS KINASE 1 73914 NM_028679 MUS INTERLEUKIN-1
RECEPTOR-ASSOCIATED MUSCULUS KINASE 3 16647 NM_010655 MUS
KARYOPHERIN (IMPORTIN) ALPHA 2 MUSCULUS 16542 NM_010612 MUS SP.
KINASE INSERT DOMAIN PROTEIN RECEPTOR 16542 NM_010612 MUS KINASE
INSERT DOMAIN PROTEIN MUSCULUS RECEPTOR 17347 NM_021462 MUS MAP
KINASE-INTERACTING MUSCULUS SERINE/THREONINE KINASE 2 226778
NM_145515 MUS MAP/MICROTUBULE AFFINITY- MUSCULUS REGULATING KINASE
1 13384 NM_007863 MUS MEMBRANE PROTEIN, PALMITOYLATED 3 MUSCULUS
(MAGUK P55 SUBFAMILY MEMBER 3) 17295 NM_008591 MUS MET
PROTO-ONCOGENE MUSCULUS 17776 NM_008641 MUS MICROTUBULE ASSOCIATED
MUSCULUS SERINE/THREONINE KINASE 2 26414 NM_009158 MUS MITOGEN
ACTIVATED PROTEIN KINASE MUSCULUS 10 26416 NM_011951 MUS MITOGEN
ACTIVATED PROTEIN KINASE MUSCULUS 14 26419 NM_016700 MUS MITOGEN
ACTIVATED PROTEIN KINASE 8 MUSCULUS 26395 NM_008927 MUS MITOGEN
ACTIVATED PROTEIN KINASE MUSCULUS KINASE 1 26403 NM_022012 MUS
MITOGEN ACTIVATED PROTEIN KINASE MUSCULUS KINASE KINASE 11 26404
NM_009582 MUS MITOGEN ACTIVATED PROTEIN KINASE MUSCULUS KINASE
KINASE 12 26407 NM_011948 MUS MITOGEN ACTIVATED PROTEIN KINASE
MUSCULUS KINASE KINASE 4 26410 NM_007746 MUS MITOGEN ACTIVATED
PROTEIN KINASE MUSCULUS KINASE KINASE 8 18198 NM_001037128 MUS
MUSCLE, SKELETAL, RECEPTOR MUSCULUS TYROSINE KINASE 18198 NM_010944
MUS MUSCLE, SKELETAL, RECEPTOR MUSCULUS TYROSINE KINASE 18198
NM_001037127 MUS MUSCLE, SKELETAL, RECEPTOR MUSCULUS TYROSINE
KINASE 18198 NM_001037129 MUS MUSCLE, SKELETAL, RECEPTOR MUSCULUS
TYROSINE KINASE 18198 NM_001037130 MUS MUSCLE, SKELETAL, RECEPTOR
MUSCULUS TYROSINE KINASE 23954 NM_011848 MUS NIMA (NEVER IN MITOSIS
GENE A)- MUSCULUS RELATED EXPRESSED KINASE 3 330721 NM_177898 MUS
NIMA (NEVER IN MITOSIS GENE A)- MUSCULUS RELATED EXPRESSED KINASE 5
18457 NM_019788 MUS PALLIDIN MUSCULUS 269614 NM_172990 MUS
PANTOTHENATE KINASE 4 MUSCULUS 18707 NM_001029837 MUS
PHOSPHATIDYLINOSITOL 3-KINASE MUSCULUS CATALYTIC DELTA POLYPEPTIDE
18707 NM_008840 MUS PHOSPHATIDYLINOSITOL 3-KINASE MUSCULUS
CATALYTIC DELTA POLYPEPTIDE 74769 NM_029094 MUS
PHOSPHATIDYLINOSITOL 3-KINASE, MUSCULUS CATALYTIC, BETA POLYPEPTIDE
18720 NM_008847 MUS PHOSPHATIDYLINOSITOL-4-PHOSPHATE 5- MUSCULUS
KINASE, TYPE 1 BETA 18641 NM_008826 MUS PHOSPHOFRUCTOKINASE, LIVER,
B-TYPE MUSCULUS 18655 NM_008828 MUS PHOSPHOGLYCERATE KINASE 1
MUSCULUS 18655 XM_484116 MUS PHOSPHOGLYCERATE KINASE 1 MUSCULUS
18655 XM_485239 MUS PHOSPHOGLYCERATE KINASE 1 MUSCULUS 225326
NM_181414 MUS PHOSPHOINOSITIDE-3-KINASE, CLASS 3 MUSCULUS 18595
NM_011058 MUS PLATELET DERIVED GROWTH FACTOR MUSCULUS RECEPTOR,
ALPHA POLYPEPTIDE 20620 NM_152804 MUS POLO-LIKE KINASE 2
(DROSOPHILA) MUSCULUS 234852 NM_145606 MUS PROCOLLAGEN (TYPE III)
N- MUSCULUS ENDOPEPTIDASE 18762 NM_001039079 MUS PROTEIN KINASE C,
ZETA MUSCULUS 18762 NM_008860 MUS PROTEIN KINASE C, ZETA MUSCULUS
18747 NM_008854 MUS PROTEIN KINASE, CAMP DEPENDENT, MUSCULUS
CATALYTIC, ALPHA 18712 NM_008842 MUS PROVIRAL INTEGRATION SITE 1
MUSCULUS 68708 NM_026817 MUS RAB, MEMBER OF RAS ONCOGENE MUSCULUS
FAMILY-LIKE 2A 59021 NM_021518 MUS RAB2, MEMBER RAS ONCOGENE FAMILY
MUSCULUS 19336 NM_009000 MUS RAB24, MEMBER RAS ONCOGENE FAMILY
MUSCULUS 76338 NM_172601 MUS RAB2B, MEMBER RAS ONCOGENE FAMILY
MUSCULUS 19376 NM_033475 MUS SP. RAB34, MEMBER OF RAS ONCOGENE
FAMILY 19376 NM_033475 MUS RAB34, MEMBER OF RAS ONCOGENE MUSCULUS
FAMILY 67790 NM_175122 MUS RAB39B, MEMBER RAS ONCOGENE MUSCULUS
FAMILY 226422 NM_144875 MUS RAB7, MEMBER RAS ONCOGENE FAMILY-
MUSCULUS LIKE 1
56382 NM_019773 MUS RAB9, MEMBER RAS ONCOGENE FAMILY MUSCULUS 11848
NM_016802 MUS RAS HOMOLOG GENE FAMILY, MEMBER A MUSCULUS 23912
NM_175092 MUS RAS HOMOLOG GENE FAMILY, MEMBER F MUSCULUS 74734
NM_001081105 MUS RAS HOMOLOG GENE FAMILY, MEMBER H XM_132051
MUSCULUS 74734 XM_903893 MUS RAS HOMOLOG GENE FAMILY, MEMBER H
MUSCULUS 74734 XM_903680 MUS RAS HOMOLOG GENE FAMILY, MEMBER H
MUSCULUS 74734 XM_924029 MUS RAS HOMOLOG GENE FAMILY, MEMBER H
MUSCULUS 74734 XM_622908 MUS RAS HOMOLOG GENE FAMILY, MEMBER H
MUSCULUS 74734 XM_924031 MUS RAS HOMOLOG GENE FAMILY, MEMBER H
MUSCULUS 74734 XM_915950 MUS RAS HOMOLOG GENE FAMILY, MEMBER H
MUSCULUS 74734 XM_900704 MUS RAS HOMOLOG GENE FAMILY, MEMBER H
MUSCULUS 74734 XM_132051 MUS RAS HOMOLOG GENE FAMILY, MEMBER H
MUSCULUS 215449 NM_024457 MUS RAS RELATED PROTEIN 1B MUSCULUS 19744
NM_053075 MUS RAS-HOMOLOG ENRICHED IN BRAIN MUSCULUS 56532
NM_019955 MUS RECEPTOR-INTERACTING SERINE- MUSCULUS THREONINE
KINASE 3 69288 NM_001081347 MUS RHO-RELATED BTB DOMAIN CONTAINING 1
XM_897555 MUSCULUS 69288 XM_897555 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_125637 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_920631 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_920652 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_897548 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_907869 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_897523 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_920622 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_897577 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_920637 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_920646 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_897586 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_920664 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_887557 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 69288 XM_920656 MUS RHO-RELATED BTB DOMAIN
CONTAINING 1 MUSCULUS 58988 NM_021485 MUS RIBOSOMAL PROTEIN S6
KINASE, MUSCULUS POLYPEPTIDE 2 231532 NM_146161 MUS RIKEN CDNA
0610025G21 GENE MUSCULUS 231532 NM_029270 MUS RIKEN CDNA 0610025G21
GENE MUSCULUS 66201 NM_025418 MUS RIKEN CDNA 1110059P08 GENE
MUSCULUS 68870 NM_001033874 MUS RIKEN CDNA 1190002A17 GENE MUSCULUS
329509 NM_198630 MUS RIKEN CDNA 1810024B03 GENE MUSCULUS 66371
NM_025519 MUS RIKEN CDNA 2310010I16 GENE MUSCULUS 75619 NM_172422
MUS RIKEN CDNA 2810421I24 GENE MUSCULUS 225028 NM_001081357 MUS
RIKEN CDNA 4833416M01 GENE XM_898848 MUSCULUS 225028 XM_898825 MUS
RIKEN CDNA 4833416M01 GENE MUSCULUS 225028 XM_898819 MUS RIKEN CDNA
4833416M01 GENE MUSCULUS 225028 XM_898848 MUS RIKEN CDNA 4833416M01
GENE MUSCULUS 225028 XM_898843 MUS RIKEN CDNA 4833416M01 GENE
MUSCULUS 225028 XM_898830 MUS RIKEN CDNA 4833416M01 GENE MUSCULUS
225028 XM_898852 MUS RIKEN CDNA 4833416M01 GENE MUSCULUS 225028
XM_898838 MUS RIKEN CDNA 4833416M01 GENE MUSCULUS 74354 XM_910825
MUS RIKEN CDNA 4921528H16 GENE MUSCULUS 74354 XM_895665 MUS RIKEN
CDNA 4921528H16 GENE MUSCULUS 74354 NM_028886 MUS LEUCINE-RICH
REPEATS AND XM_133060 MUSCULUS GUANYLATE KINASE DOMAIN XM_910825
CONTAINING (LRGUK) 74354 XM_921792 MUS RIKEN CDNA 4921528H16 GENE
MUSCULUS 211496 NM_177599 MUS RIKEN CDNA 4932415M13 GENE MUSCULUS
211496 NM_001037718 MUS RIKEN CDNA 4932415M13 GENE MUSCULUS 319613
NM_176998 MUS RIKEN CDNA 5730410E15 GENE MUSCULUS 319613
NM_001032727 MUS RIKEN CDNA 5730410E15 GENE MUSCULUS 319613
NM_178765 MUS RIKEN CDNA 5730410E15 GENE MUSCULUS 110157 NM_029780
MUS RIKEN CDNA 6430402F14 GENE MUSCULUS 215114 NM_146001 MUS RIKEN
CDNA A930014B11 GENE MUSCULUS 20224 NM_009120 MUS SAR1 GENE HOMOLOG
A (S. CEREVISIAE) MUSCULUS 76983 NM_029825 MUS SEC1 FAMILY DOMAIN
CONTAINING 1 MUSCULUS 104175 NM_145587 MUS SH3-BINDING KINASE 1
MUSCULUS 24056 NM_011894 MUS SH3-DOMAIN BINDING PROTEIN 5 (BTK-
MUSCULUS ASSOCIATED) 385049 XM_001473528 MUS SIMILAR TO
RHO-ASSOCIATED COILED- XM_904204 MUSCULUS COIL FORMING KINASE 1
385049 XM_358017 MUS SIMILAR TO RHO-ASSOCIATED COILED- MUSCULUS
COIL FORMING KINASE 1 381390 XM_485079 MUS SIMILAR TO
SERINE/THREONINE KINASE MUSCULUS 381390 XM_355352 MUS GM14147
PREDICTED GENE 14147 MUSCULUS 71982 NM_028035 MUS SORTING NEXIN 10
MUSCULUS 67804 NM_026386 MUS SORTING NEXIN 2 MUSCULUS 67727
NM_026343 MUS SYNTAXIN 17 MUSCULUS 55943 NM_018768 MUS SYNTAXIN 8
MUSCULUS 20912 NM_011504 MUS SYNTAXIN BINDING PROTEIN 3A MUSCULUS
20912 NM_198326 MUS SYNTAXIN BINDING PROTEIN 3A MUSCULUS 56480
NM_019786 MUS TANK-BINDING KINASE 1 MUSCULUS 58864 NM_080442 MUS
TESTIS-SPECIFIC SERINE KINASE 3 MUSCULUS 83984 NM_032004 MUS
TESTIS-SPECIFIC SERINE KINASE 6 MUSCULUS 66805 NM_133681 MUS
TETRASPANIN 1 MUSCULUS 70747 NM_027533 MUS TETRASPANIN 2 MUSCULUS
109246 NM_175414 MUS TETRASPANIN 9 MUSCULUS 73412 NM_181591 MUS
THIOREDOXIN DOMAIN CONTAINING 3 MUSCULUS (SPERMATOZOA) 21877
NM_009387 MUS SP. THYMIDINE KINASE 1 21877 NM_009387 MUS THYMIDINE
KINASE 1 MUSCULUS 11651 NM_009652 MUS THYMOMA VIRAL PROTO-ONCOGENE
1 MUSCULUS 24086 NM_011903 MUS TOUSLED-LIKE KINASE 2 (ARABIDOPSIS)
MUSCULUS 100683 XM_891798 MUS TRANSFORMATION/TRANSCRIPTION MUSCULUS
DOMAIN-ASSOCIATED PROTEIN 100683 XM_899747 MUS
TRANSFORMATION/TRANSCRIPTION MUSCULUS DOMAIN-ASSOCIATED PROTEIN
100683 XM_918276 MUS TRANSFORMATION/TRANSCRIPTION MUSCULUS
DOMAIN-ASSOCIATED PROTEIN 100683 NM_001081362 MUS
TRANSFORMATION/TRANSCRIPTION XM_899741 MUSCULUS DOMAIN-ASSOCIATED
PROTEIN 100683 XM_899763 MUS TRANSFORMATION/TRANSCRIPTION MUSCULUS
DOMAIN-ASSOCIATED PROTEIN 100683 XM_917315 MUS
TRANSFORMATION/TRANSCRIPTION MUSCULUS DOMAIN-ASSOCIATED PROTEIN
100683 XM_886427 MUS TRANSFORMATION/TRANSCRIPTION MUSCULUS
DOMAIN-ASSOCIATED PROTEIN 100683 XM_899754 MUS
TRANSFORMATION/TRANSCRIPTION MUSCULUS DOMAIN-ASSOCIATED PROTEIN
100683 XM_925613 MUS TRANSFORMATION/TRANSCRIPTION MUSCULUS
DOMAIN-ASSOCIATED PROTEIN 100683 XM_925614 MUS
TRANSFORMATION/TRANSCRIPTION MUSCULUS DOMAIN-ASSOCIATED PROTEIN
100683 XM_899771 MUS TRANSFORMATION/TRANSCRIPTION MUSCULUS
DOMAIN-ASSOCIATED PROTEIN 100683 XM_918275 MUS
TRANSFORMATION/TRANSCRIPTION MUSCULUS DOMAIN-ASSOCIATED PROTEIN
100683 XM_917317 MUS TRANSFORMATION/TRANSCRIPTION MUSCULUS
DOMAIN-ASSOCIATED PROTEIN 100683 XM_899733 MUS
TRANSFORMATION/TRANSCRIPTION MUSCULUS DOMAIN-ASSOCIATED PROTEIN
100683 XM_925612 MUS TRANSFORMATION/TRANSCRIPTION MUSCULUS
DOMAIN-ASSOCIATED PROTEIN 108989 NM_133780 MUS TRANSLOCATED
PROMOTER REGION MUSCULUS 17112 NM_008536 MUS TRANSMEMBRANE 4
SUPERFAMILY MUSCULUS MEMBER 1 21848 NM_145076 MUS SP. TRIPARTITE
MOTIF PROTEIN 24 21848 NM_145076 MUS TRIPARTITE MOTIF PROTEIN 24
MUSCULUS 19720 NM_009054 MUS TRIPARTITE MOTIF PROTEIN 27 MUSCULUS
13867 NM_010153 MUS V-ERB-B2 ERYTHROBLASTIC LEUKEMIA MUSCULUS VIRAL
ONCOGENE HOMOLOG 3 (AVIAN) 16653 NM_010937 MUS V-KI-RAS2 KIRSTEN
RAT SARCOMA VIRAL MUSCULUS ONCOGENE HOMOLOG 16653 NM_021284 MUS
V-KI-RAS2 KIRSTEN RAT SARCOMA VIRAL MUSCULUS ONCOGENE HOMOLOG 69922
NM_027260 MUS VACCINIA RELATED KINASE 2 MUSCULUS 233405 NM_178070
MUS VACUOLAR PROTEIN SORTING 33B MUSCULUS (YEAST) 22320 NM_016794
MUS VESICLE-ASSOCIATED MEMBRANE MUSCULUS PROTEIN 8 30960 NM_013933
MUS VESICLE-ASSOCIATED MEMBRANE MUSCULUS PROTEIN, ASSOCIATED
PROTEIN A 218456 N/A MUS SIMILAR TO NUCLEOSIDE DIPHOSPHATE MUSCULUS
KINASE B (NDK B) (NDP KINASE B) (P18 381082 N/A MUS SIMILAR TO
MITOGEN-ACTIVATED MUSCULUS PROTEIN KINASE 14 ISOFORM 1; CYTOKINE
SUP 381309 N/A MUS SIMILAR TO CDC42-BINDING PROTEIN MUSCULUS KINASE
ALPHA; MYTONIC DYSTROPHY KINAS 384481 N/A MUS SIMILAR TO URIDINE
MONOPHOSPHATE MUSCULUS KINASE
Example 2
[0089] Table 2 shows the results of a screen that led to
accumulation of p62. The results are presented in a positive to
negative ranking according to p62.
TABLE-US-00003 TABLE 2 Symbol Gene ID Gene rank Chmp4b 75608 1
Kpnb1 16211 2 Irak1 16179 3 Rhov 228543 4 Gm5285 383956 5 Eef1a2
13628 6 Ulk1 22241 7 Epha5 13839 8 LUCIFERASE -14 9 Rhot1 59040 10
Stk35 67333 11 Gtpbp4 69237 12 Prkaa2 108079 13 Pfkfb4 270198 14
Map3k14 53859 15 Rhog 56212 16 Mylk4 238564 17 Rhoc 11853 18
Cdc42bpg 240505 19 GeneID: 245619 245619 20 Tssk3 58864 21 Ap1s1
11769 22 Smok2a 27263 23 Agap3 213990 24 Shpk 74637 25 Cdk3 69681
26 N4bp2 333789 27 Ap3m2 64933 28 Rab20 19332 29 Vps4b 20479 30 Hgs
15239 31 Pik3c2a 18704 32 Etnk1 75320 33 Sh3gl2 20404 34 Prkar2b
19088 35 Rac1 19353 36 Gpn1 74254 37 Nkiras2 71966 38 Pik3cg 30955
39 Prkag1 19082 40 Phkg1 18682 41 GFP -10 42 Vps13a 271564 43 Txk
22165 44 Dab2 13132 45 Mast3 234385 46 Pdgfrl 68797 47 Vps36 70160
48 GeneID: 233024 #N/A 49 Prkaa1 105787 50 Pdpk1 18607 51 Doc2b
13447 52 Gm5374 385049 53 Nuak1 77976 54 Acvr2a 11480 55 Snx3 54198
56 Mapk9 26420 57 Camk4 12326 58 Grk6 26385 59 Nme2 18103 60 Pak4
70584 61 Stradb 227154 62 Rab19 19331 63 Pi4ka 224020 64 Camk2g
12325 65 Dlg4 13385 66 Cdadc1 71891 67 Mapkapk3 102626 68 Tjp2
21873 69 Gm318 240091 70 Cdkl5 382253 71 Snx16 74718 72 Cdc2l1
12537 73 Trpm6 225997 74 Ap1g1 11765 75 Rab14 68365 76 RFP -12 77
Pmvk 68603 78 Pank3 211347 79 Pkmyt1 268930 80 Crkl 12929 81
Fastkd5 380601 82 Etnk2 214253 83 Ephb6 13848 84 GeneID: 229309
#N/A 85 Aldh18a1 56454 86 Ak1 11636 87 Eif2ak1 15467 88 Kdr 16542
89 Gsk3a 606496 90 Gck 103988 91 Araf 11836 92 Arf5 11844 93 Dgkb
217480 94 Ltk 17005 95 Rab8a 17274 96 Ryk 20187 97 Gm4862 229879 98
Mknk2 17347 99 GeneID: 243968 #N/A 100 Arpc1a 56443 101
2310079N02Rik 66566 102 GeneID: 381981 #N/A 103 Nme7 171567 104 Ak2
11637 105 Tesk2 230661 106 Sik1 17691 107 Synj2 20975 108 Chek1
12649 109 GeneID: 384257 #N/A 110 Mapk3 26417 111 Itpka 228550 112
Gm1078 381835 113 Ciita 12265 114 GeneID: 381061 #N/A 115 Uck1
22245 116 Rps6ka2 20112 117 Prps111 75456 118 Rap1a 109905 119 Rac3
170758 120 Gm4776 212225 121 Eif2s3y 26908 122 Trpd5213 66745 123
GeneID: 245068 #N/A 124 lacZ -15 125 Sec23a 20334 126 Arfrp1 76688
127 Ras110b 276952 128 Hspb8 80888 129 Rab3d 19340 130 Arl4d 80981
131 GeneID: 384894 #N/A 132 GeneID: 381446 #N/A 133 GeneID: 268321
#N/A 134 Anxa7 11750 135 Hras1 15461 136 Wnk4 69847 137 Melk 17279
138 Frk 14302 139 Mapkapk2 17164 140 Rragc 54170 141 Phka1 18679
142 Becn1 56208 143 Pik3c2g 18705 144 Acvr2b 11481 145 Tie1 21846
146 Camk1g 215303 147 Phkg2 68961 148 Csnk1g1 214897 149 Khk 16548
150 Fgfr4 14186 151 Gucy2e 14919 152 Gm1893 381599 153 Rras2 66922
154 Ikbkb 16150 155 Gsg2 14841 156 Tspan7 21912 157 Keap1 50868 158
Rab6b 270192 159 Ern1 78943 160 Golga5 27277 161 Mlkl 74568 162
Epha6 13840 163 Ephb2 13844 164 Camkk1 55984 165 Psmc1 19179 166
Ankk1 244859 167 B2m 12010 168 Nkiras1 69721 169 Rab15 104886 170
Smg1 233789 171 Ckmt2 76722 172 Gm9824 432447 173 Nek4 23955 174
Trp53rk 76367 175 Mtor 56717 176 Vps39 269338 177 Rps6kl1 238323
178 Drg2 13495 179 Mark4 232944 180 Syt15 319508 181 Rerg 232441
182 Hipk2 15258 183 Cav3 12391 184 Grk5 14773 185 Map2k3 26397 186
Smok4a 272667 187 Rab7 19349 188 2810408M09Rik 381406 189 Chmp5
76959 190 Prkcd 18753 191 Snx2 67804 192 Mst1r 19882 193 Stk19
54402 194 Gk2 14626 195 Acvrl1 11482 196 Syt8 55925 197 Cyth3 19159
198 Tspan6 56496 199 Irak3 73914 200
TABLE-US-00004 TABLE 4 GeneID RefSeq Species Description 18607
NM_011062 MUS 3-PHOSPHOINOSITIDE DEPENDENT MUSCULUS PROTEIN
KINASE-1 270198 NM_173019 MUS 6-PHOSPHOFRUCTO-2- MUSCULUS
KINASE/FRUCTOSE-2,6- BIPHOSPHATASE 4 270198 NM_001039217 MUS
6-PHOSPHOFRUCTO-2- MUSCULUS KINASE/FRUCTOSE-2,6- BIPHOSPHATASE 4
270198 NM_001039216 MUS 6-PHOSPHOFRUCTO-2- MUSCULUS
KINASE/FRUCTOSE-2,6- BIPHOSPHATASE 4 270198 NM_001039215 MUS
6-PHOSPHOFRUCTO-2- MUSCULUS KINASE/FRUCTOSE-2,6- BIPHOSPHATASE 4
56443 NM_019767 MUS ACTIN RELATED PROTEIN 2/3 MUSCULUS COMPLEX,
SUBUNIT 1A 11482 NM_009612 MUS ACTIVIN A RECEPTOR, TYPE II-LIKE 1
MUSCULUS 11480 NM_007396 MUS ACTIVIN RECEPTOR IIA MUSCULUS 11481
NM_007397 MUS ACTIVIN RECEPTOR IIB MUSCULUS 11765 NM_009677 MUS
ADAPTOR PROTEIN COMPLEX AP-1, MUSCULUS GAMMA 1 SUBUNIT 11769
NM_007457 MUS ADAPTOR PROTEIN COMPLEX AP-1, MUSCULUS SIGMA 1 64933
NM_029505 MUS ADAPTOR-RELATED PROTEIN MUSCULUS COMPLEX 3, MU 2
SUBUNIT 11636 NM_021515 MUS ADENYLATE KINASE 1 MUSCULUS 11637
NM_001033966 MUS ADENYLATE KINASE 2 MUSCULUS 11637 NM_016895 MUS
ADENYLATE KINASE 2 MUSCULUS 11844 NM_007480 MUS ADP-RIBOSYLATION
FACTOR 5 MUSCULUS 76688 NM_029702 MUS ADP-RIBOSYLATION FACTOR
MUSCULUS RELATED PROTEIN 1 80981 NM_031160 MUS ADP-RIBOSYLATION
FACTOR-LIKE 4D MUSCULUS 56454 NM_019698 MUS ALDEHYDE DEHYDROGENASE
18 MUSCULUS FAMILY, MEMBER A1 56454 NM_153554 MUS ALDEHYDE
DEHYDROGENASE 18 MUSCULUS FAMILY, MEMBER A1 227154 NM_172656 MUS
AMYOTROPHIC LATERAL SCLEROSIS 2 MUSCULUS (JUVENILE) CHROMOSOME
REGION, CANDIDAT . . . 244859 NM_172922 MUS ANKYRIN REPEAT AND
KINASE MUSCULUS DOMAIN CONTAINING 1 11750 NM_009674 MUS ANNEXIN A7
MUSCULUS 333789 NM_001024917 MUS BCL3 BINDING PROTEIN MUSCULUS
56208 NM_019584 MUS BECLIN 1 (COILED-COIL, MYOSIN-LIKE MUSCULUS
BCL2-INTERACTING PROTEIN) 12010 NM_009735 MUS BETA-2 MICROGLOBULIN
SPRETUS 12010 NM_009735 MUS BETA-2 MICROGLOBULIN MUSCULUS 215303
NM_144817 MUS CALCIUM/CALMODULIN-DEPENDENT MUSCULUS PROTEIN KINASE
I GAMMA 12325 NM_178597 MUS CALCIUM/CALMODULIN-DEPENDENT MUSCULUS
PROTEIN KINASE II GAMMA 12325 NM_001039139 MUS
CALCIUM/CALMODULIN-DEPENDENT MUSCULUS PROTEIN KINASE II GAMMA 12325
NM_001039138 MUS CALCIUM/CALMODULIN-DEPENDENT MUSCULUS PROTEIN
KINASE II GAMMA 12326 NM_009793 MUS CALCIUM/CALMODULIN-DEPENDENT
MUSCULUS PROTEIN KINASE IV 55984 NM_018883 MUS
CALCIUM/CALMODULIN-DEPENDENT MUSCULUS PROTEIN KINASE KINASE 1,
ALPHA 74637 NM_029031 MUS CARBOHYDRATE KINASE-LIKE MUSCULUS 214897
NM_173185 MUS CASEIN KINASE 1, GAMMA 1 MUSCULUS 12391 NM_007617 MUS
CAVEOLIN 3 MUSCULUS 240505 NM_001033342 MUS CDC42 BINDING PROTEIN
KINASE XM_906449 MUSCULUS GAMMA (DMPK-LIKE) 240505 NM_001033342 MUS
CDC42 BINDING PROTEIN KINASE XM_140553 MUSCULUS GAMMA (DMPK-LIKE)
12537 NM_007661 MUS CELL DIVISION CYCLE 2 HOMOLOG (S. POMBE)-
MUSCULUS LIKE 1 213990 NM_139153 MUS CENTAURIN, GAMMA 3 MUSCULUS
12649 NM_007691 MUS CHECKPOINT KINASE 1 HOMOLOG (S. POMBE) MUSCULUS
13848 NM_007680 MUS CHICKEN EPH/ELK RECEPTOR-LIKE MUSCULUS PROTEIN
75608 NM_029362 MUS CHROMATIN MODIFYING PROTEIN 4B MUSCULUS 76959
NM_029814 MUS CHROMATIN MODIFYING PROTEIN 5 MUSCULUS 12265
NM_007575 MUS CLASS II TRANSACTIVATOR MUSCULUS 76722 NM_198415 MUS
CREATINE KINASE, MITOCHONDRIAL 2 MUSCULUS 69681 NM_027165 MUS
CYCLIN-DEPENDENT KINASE 3 MUSCULUS 382253 XM_912167 MUS
CYCLIN-DEPENDENT KINASE-LIKE 5 NM_001024624 MUSCULUS NM_027986
XM_914101 XM_001000245 XM_001000259 XM_001000276 XM_001000293 71891
XM_001000308 MUS CYTIDINE AND DCMP DEAMINASE XM_001000321 MUSCULUS
DOMAIN CONTAINING 1 XM_001000336 XM_001002774 XM_127813 XM_914101
XM_985192 71891 XM_901860 MUS CYTIDINE AND DCMP DEAMINASE MUSCULUS
DOMAIN CONTAINING 1 71891 XM_923095 MUS CYTIDINE AND DCMP DEAMINASE
MUSCULUS DOMAIN CONTAINING 1 71891 XM_894723 MUS CYTIDINE AND DCMP
DEAMINASE MUSCULUS DOMAIN CONTAINING 1 71891 XM_923089 MUS CYTIDINE
AND DCMP DEAMINASE MUSCULUS DOMAIN CONTAINING 1 71891 XM_901847 MUS
CYTIDINE AND DCMP DEAMINASE MUSCULUS DOMAIN CONTAINING 1 71891
XM_901853 MUS CYTIDINE AND DCMP DEAMINASE MUSCULUS DOMAIN
CONTAINING 1 71891 XM_923082 MUS CYTIDINE AND DCMP DEAMINASE
MUSCULUS DOMAIN CONTAINING 1 71891 XM_923072 MUS CYTIDINE AND DCMP
DEAMINASE MUSCULUS DOMAIN CONTAINING 1 71891 XM_923086 MUS CYTIDINE
AND DCMP DEAMINASE MUSCULUS DOMAIN CONTAINING 1 71891 XM_901857 MUS
CYTIDINE AND DCMP DEAMINASE MUSCULUS DOMAIN CONTAINING 1 71891
XM_127813 MUS CYTIDINE AND DCMP DEAMINASE MUSCULUS DOMAIN
CONTAINING 1 71891 XM_901866 MUS CYTIDINE AND DCMP DEAMINASE
MUSCULUS DOMAIN CONTAINING 1 71891 XM_923093 MUS CYTIDINE AND DCMP
DEAMINASE MUSCULUS DOMAIN CONTAINING 1 13495 NM_021354 MUS
DEVELOPMENTALLY REGULATED GTP MUSCULUS BINDING PROTEIN 2 217480
NM_178681 MUS DIACYLGLYCEROL KINASE, BETA MUSCULUS 13132
NM_001037905 MUS DISABLED HOMOLOG 2 (DROSOPHILA) MUSCULUS 13132
NM_001008702 MUS DISABLED HOMOLOG 2 (DROSOPHILA) MUSCULUS 13132
NM_023118 MUS DISABLED HOMOLOG 2 (DROSOPHILA) MUSCULUS 13385
NM_007864 MUS DISCS, LARGE HOMOLOG 4 MUSCULUS (DROSOPHILA) 13447
NM_007873 MUS DOUBLE C2, BETA MUSCULUS 13839 NM_007937 MUS ECK-LIKE
SEQUENCE 1 MUSCULUS 78943 NM_023913 MUS ENDOPLASMIC RETICULUM (ER)
TO MUSCULUS NUCLEUS SIGNALLING 1 13840 NM_007938 MUS EPH RECEPTOR
A6 MUSCULUS 13844 NM_010142 MUS EPH RECEPTOR B2 MUSCULUS 75320
XM_284250 MUS ETHANOLAMINE KINASE 1 MUSCULUS 75320 NM_029250 MUS
ETHANOLAMINE KINASE 1 XM_908334 MUSCULUS XM_979562 214253 NM_175443
MUS ETHANOLAMINE KINASE 2 MUSCULUS 13628 NM_007906 MUS EUKARYOTIC
TRANSLATION MUSCULUS ELONGATION FACTOR 1 ALPHA 2 15467 NM_013557
MUS EUKARYOTIC TRANSLATION MUSCULUS INITIATION FACTOR 2 ALPHA
KINASE 1 26908 NM_012011 MUS EUKARYOTIC TRANSLATION MUSCULUS
INITIATION FACTOR 2, SUBUNIT 3, STRUCTURAL GENE . . . 18103
NM_008705 MUS EXPRESSED IN NON-METASTATIC MUSCULUS CELLS 2, PROTEIN
380601 NM_198176 MUS EXPRESSED SEQUENCE C78212 MUSCULUS 14186
NM_008011 MUS FIBROBLAST GROWTH FACTOR MUSCULUS RECEPTOR 4 56717
NM_020009 MUS FK506 BINDING PROTEIN 12- MUSCULUS RAPAMYCIN
ASSOCIATED PROTEIN 1 56717 NM_001039554 MUS FK506 BINDING PROTEIN
12- MUSCULUS RAPAMYCIN ASSOCIATED PROTEIN 1 14302 NM_010237 MUS
FYN-RELATED KINASE MUSCULUS 14773 NM_018869 MUS G PROTEIN-COUPLED
RECEPTOR MUSCULUS KINASE 5 26385 NM_011938 MUS G PROTEIN-COUPLED
RECEPTOR MUSCULUS KINASE 6 26385 NM_001038018 MUS G PROTEIN-COUPLED
RECEPTOR MUSCULUS KINASE 6 381835 XM_355840 MUS GENE MODEL 1078,
(NCBI) MUSCULUS 381835 XM_911944 MUS GENE MODEL 1078, (NCBI)
MUSCULUS 381599 XM_355556 MUS GENE MODEL 1893, (NCBI) MUSCULUS
240091 XM_139919 MUS GENE MODEL 318, (NCBI) [replaced XM_619462
MUSCULUS with XM_001481303 545204] XM_622850 14841 NM_010353 MUS
GERM CELL-SPECIFIC GENE 2 MUSCULUS 103988 NM_010292 MUS GLUCOKINASE
MUSCULUS 14626 NM_010294 MUS GLYCEROL KINASE 2 MUSCULUS 606496
NM_001031667 MUS GLYCOGEN SYNTHASE KINASE 3 MUSCULUS ALPHA 27277
NM_013747 MUS GOLGI AUTOANTIGEN, GOLGIN MUSCULUS SUBFAMILY A, 5
69237 NM_027000 MUS GTP BINDING PROTEIN 4 MUSCULUS 14919 NM_008192
MUS GUANYLATE CYCLASE 2E MUSCULUS 15461 NM_008284 MUS SP. HARVEY
RAT SARCOMA VIRUS ONCOGENE 1 15461 NM_008284 MUS HARVEY RAT SARCOMA
VIRUS MUSCULUS ONCOGENE 1 80888 NM_030704 MUS HEAT SHOCK PROTEIN 8
MUSCULUS 15239 NM_008244 MUS HGF-REGULATED TYROSINE KINASE MUSCULUS
SUBSTRATE 15258 NM_010433 MUS HOMEODOMAIN INTERACTING MUSCULUS
PROTEIN KINASE 2 212225 NM_172504 MUS HYPOTHETICAL PROTEIN
4930509O22 MUSCULUS 16150 NM_010546 MUS INHIBITOR OF KAPPAB KINASE
BETA MUSCULUS 228550 NM_146125 MUS INOSITOL 1,4,5-TRISPHOSPHATE 3-
MUSCULUS KINASE A 16179 NM_008363 MUS INTERLEUKIN-1 RECEPTOR-
MUSCULUS ASSOCIATED KINASE 1 73914 NM_028679 MUS INTERLEUKIN-1
RECEPTOR- MUSCULUS ASSOCIATED KINASE 3 16211 NM_008379 MUS
KARYOPHERIN (IMPORTIN) BETA 1 MUSCULUS 50868 NM_016679 MUS
KELCH-LIKE ECH-ASSOCIATED MUSCULUS PROTEIN 1 16548 NM_008439 MUS
KETOHEXOKINASE MUSCULUS 16542 NM_010612 MUS KINASE INSERT DOMAIN
PROTEIN
MUSCULUS RECEPTOR 16542 NM_010612 MUS SP. KINASE INSERT DOMAIN
PROTEIN RECEPTOR 17005 NM_008523 MUS LEUKOCYTE TYROSINE KINASE
MUSCULUS 17005 NM_206942 MUS LEUKOCYTE TYROSINE KINASE MUSCULUS
17005 NM_206941 MUS LEUKOCYTE TYROSINE KINASE MUSCULUS 17005
NM_203345 MUS LEUKOCYTE TYROSINE KINASE MUSCULUS 19882 NM_009074
MUS SP. MACROPHAGE STIMULATING 1 RECEPTOR (C-MET-RELATED TYROSINE
KINASE) 19882 NM_009074 MUS MACROPHAGE STIMULATING 1 MUSCULUS
RECEPTOR (C-MET-RELATED TYROSINE KINASE) 17164 NM_008551 MUS MAP
KINASE-ACTIVATED PROTEIN MUSCULUS KINASE 2 17347 NM_021462 MUS MAP
KINASE-INTERACTING MUSCULUS SERINE/THREONINE KINASE 2 232944
NM_172279 MUS MAP/MICROTUBULE AFFINITY- MUSCULUS REGULATING KINASE
4 17279 NM_010790 MUS MATERNAL EMBRYONIC LEUCINE MUSCULUS ZIPPER
KINASE 268930 NM_023058 MUS MEMBRANE-ASSOCIATED TYROSINE- MUSCULUS
AND THREONINE-SPECIFIC CDC2- INHIBITORY KI . . . 234385 NM_199308
MUS MICROTUBULE ASSOCIATED [replaced XM_134245 MUSCULUS
SERINE/THREONINE KINASE 3 with XM_913385 546071] 234385 XM_888290
MUS MICROTUBULE ASSOCIATED MUSCULUS SERINE/THREONINE KINASE 3
234385 XM_922751 MUS MICROTUBULE ASSOCIATED MUSCULUS
SERINE/THREONINE KINASE 3 234385 XM_897983 MUS MICROTUBULE
ASSOCIATED MUSCULUS SERINE/THREONINE KINASE 3 234385 XM_897989 MUS
MICROTUBULE ASSOCIATED MUSCULUS SERINE/THREONINE KINASE 3 234385
XM_620670 MUS MICROTUBULE ASSOCIATED MUSCULUS SERINE/THREONINE
KINASE 3 234385 XM_897962 MUS MICROTUBULE ASSOCIATED MUSCULUS
SERINE/THREONINE KINASE 3 234385 XM_897954 MUS MICROTUBULE
ASSOCIATED MUSCULUS SERINE/THREONINE KINASE 3 26420 NM_016961 MUS
MITOGEN ACTIVATED PROTEIN MUSCULUS KINASE 9 26420 NM_207692 MUS
MITOGEN ACTIVATED PROTEIN MUSCULUS KINASE 9 26397 NM_008928 MUS
MITOGEN ACTIVATED PROTEIN MUSCULUS KINASE KINASE 3 53859 NM_016896
MUS MITOGEN-ACTIVATED PROTEIN MUSCULUS KINASE KINASE KINASE 14
102626 NM_178907 MUS MITOGEN-ACTIVATED PROTEIN MUSCULUS
KINASE-ACTIVATED PROTEIN KINASE 3 74568 NM_029005 MUS MIXED LINEAGE
KINASE DOMAIN- XM_001003995 MUSCULUS LIKE XM_001003998 XM_356104
XM_895027 XM_916960 XM_924589 XM_924589 74568 XM_895027 MUS MIXED
LINEAGE KINASE DOMAIN- MUSCULUS LIKE 74568 XM_902435 MUS MIXED
LINEAGE KINASE DOMAIN- MUSCULUS LIKE 74568 XM_916960 MUS MIXED
LINEAGE KINASE DOMAIN- MUSCULUS LIKE 74568 XM_356104 MUS MIXED
LINEAGE KINASE DOMAIN- MUSCULUS LIKE 74568 XM_924585 MUS MIXED
LINEAGE KINASE DOMAIN- MUSCULUS LIKE 69721 NM_023526 MUS NFKB
INHIBITOR INTERACTING RAS- MUSCULUS LIKE PROTEIN 1 71966 NM_028024
MUS NFKB INHIBITOR INTERACTING RAS- MUSCULUS LIKE PROTEIN 2 23955
NM_011849 MUS NIMA (NEVER IN MITOSIS GENE A)- MUSCULUS RELATED
EXPRESSED KINASE 4 171567 NM_178071 MUS NON-METASTATIC CELLS 7,
PROTEIN MUSCULUS EXPRESSED IN 171567 NM_138314 MUS NON-METASTATIC
CELLS 7, PROTEIN MUSCULUS EXPRESSED IN 70584 NM_027470 MUS P21
(CDKN1A)-ACTIVATED KINASE 4 MUSCULUS 211347 NM_145962 MUS
PANTOTHENATE KINASE 3 MUSCULUS 18704 NM_011083 MUS
PHOSPHATIDYLINOSITOL 3-KINASE, MUSCULUS C2 DOMAIN CONTAINING, ALPHA
POLYPEPTIDE 18705 NM_207683 MUS PHOSPHATIDYLINOSITOL 3-KINASE,
MUSCULUS C2 DOMAIN CONTAINING, GAMMA POLYPEPTIDE 18705 NM_011084
MUS PHOSPHATIDYLINOSITOL 3-KINASE, MUSCULUS C2 DOMAIN CONTAINING,
GAMMA POLYPEPTIDE 224020 NM_001001983 MUS PHOSPHATIDYLINOSITOL
4-KINASE, MUSCULUS CATALYTIC, ALPHA POLYPEPTIDE 30955 NM_020272 MUS
PHOSPHOINOSITIDE-3-KINASE, MUSCULUS CATALYTIC, GAMMA POLYPEPTIDE
68603 NM_026784 MUS PHOSPHOMEVALONATE KINASE MUSCULUS 18679
NM_008832 MUS PHOSPHORYLASE KINASE ALPHA 1 MUSCULUS 18679 NM_173021
MUS PHOSPHORYLASE KINASE ALPHA 1 MUSCULUS 18682 NM_011079 MUS
PHOSPHORYLASE KINASE GAMMA 1 MUSCULUS 68961 NM_026888 MUS
PHOSPHORYLASE KINASE, GAMMA 2 MUSCULUS (TESTIS) 68797 NM_026840 MUS
PLATELET-DERIVED GROWTH FACTOR MUSCULUS RECEPTOR-LIKE 19159
NM_011182 MUS PLECKSTRIN HOMOLOGY, SEC7 AND MUSCULUS COILED-COIL
DOMAINS 3 19179 NM_008947 MUS PROTEASE (PROSOME, MACROPAIN)
MUSCULUS 26S SUBUNIT, ATPASE 1 18753 NM_011103 MUS PROTEIN KINASE
C, DELTA MUSCULUS 105787 NM_001013367 MUS PROTEIN KINASE,
AMP-ACTIVATED, MUSCULUS ALPHA 1 CATALYTIC SUBUNIT 108079 NM_178143
MUS PROTEIN KINASE, AMP-ACTIVATED, MUSCULUS ALPHA 2 CATALYTIC
SUBUNIT 19082 NM_016781 MUS PROTEIN KINASE, AMP-ACTIVATED, MUSCULUS
GAMMA 1 NON-CATALYTIC SUBUNIT 19088 NM_011158 MUS PROTEIN KINASE,
CAMP DEPENDENT MUSCULUS REGULATORY, TYPE II BETA 26417 NM_011952
MUS SP. PROTEIN KINASE, MITOGEN ACTIVATED KINASE 3 26417 NM_011952
MUS PROTEIN KINASE, MITOGEN MUSCULUS ACTIVATED KINASE 3 68365
NM_026697 MUS RAB14, MEMBER RAS ONCOGENE MUSCULUS FAMILY 104886
NM_134050 MUS RAB15, MEMBER RAS ONCOGENE MUSCULUS FAMILY 19331
NM_011226 MUS RAB19, MEMBER RAS ONCOGENE MUSCULUS FAMILY 19332
NM_011227 MUS RAB20, MEMBER RAS ONCOGENE MUSCULUS FAMILY 19340
NM_031874 MUS RAB3D, MEMBER RAS ONCOGENE MUSCULUS FAMILY 270192
NM_173781 MUS RAB6B, MEMBER RAS ONCOGENE MUSCULUS FAMILY 19349
NM_009005 MUS RAB7, MEMBER RAS ONCOGENE MUSCULUS FAMILY 17274
NM_023126 MUS SP. RAB8A, MEMBER RAS ONCOGENE FAMILY 17274 NM_023126
MUS RAB8A, MEMBER RAS ONCOGENE MUSCULUS FAMILY 11853 NM_007484 MUS
RAS HOMOLOG GENE FAMILY, MUSCULUS MEMBER C 56212 NM_019566 MUS RAS
HOMOLOG GENE FAMILY, MUSCULUS MEMBER G 59040 NM_021536 MUS RAS
HOMOLOG GENE FAMILY, MUSCULUS MEMBER T1 228543 NM_145530 MUS RAS
HOMOLOG GENE FAMILY, MUSCULUS MEMBER V 232441 NM_181988 MUS
RAS-LIKE, ESTROGEN-REGULATED, MUSCULUS GROWTH-INHIBITOR 276952
NM_001013386 MUS RAS-LIKE, FAMILY 10, MEMBER B MUSCULUS 19353
NM_009007 MUS RAS-RELATED C3 BOTULINUM MUSCULUS SUBSTRATE 1 170758
NM_133223 MUS RAS-RELATED C3 BOTULINUM MUSCULUS SUBSTRATE 3 54170
NM_017475 MUS RAS-RELATED GTP BINDING C MUSCULUS 109905 NM_145541
MUS RAS-RELATED PROTEIN-1A MUSCULUS 20187 NM_013649 MUS SP.
RECEPTOR-LIKE TYROSINE KINASE 20187 NM_013649 MUS RECEPTOR-LIKE
TYROSINE KINASE MUSCULUS 66922 NM_025846 MUS RELATED RAS VIRAL
(R-RAS) MUSCULUS ONCOGENE HOMOLOG 2 20112 NM_011299 MUS RIBOSOMAL
PROTEIN S6 KINASE, MUSCULUS RELATED SEQUENCE 1 238323 NM_146244 MUS
RIBOSOMAL PROTEIN S6 KINASE-LIKE 1 MUSCULUS 75456 NM_029294 MUS
RIKEN CDNA 1700011K15 GENE MUSCULUS 66566 NM_025636 MUS RIKEN CDNA
2310079N02 GENE MUSCULUS 233789 NM_001031814 MUS RIKEN CDNA
2610207I05 GENE MUSCULUS 272667 XM_895217 MUS RIKEN CDNA 4930513D10
GENE MUSCULUS 272667 XM_142762 MUS RIKEN CDNA 4930513D10 GENE
MUSCULUS 272667 XM_912174 MUS RIKEN CDNA 4930513D10 GENE MUSCULUS
271564 NM_173028 MUS RIKEN CDNA 4930516E05 GENE MUSCULUS 20334
NM_009147 MUS SEC23A (S. CEREVISIAE) MUSCULUS 54402 NM_019442 MUS
SERINE/THREONINE KINASE 19 MUSCULUS 67333 NM_001038635 MUS
SERINE/THREONINE KINASE 35 MUSCULUS 67333 NM_183262 MUS
SERINE/THREONINE KINASE 35 MUSCULUS 20404 NM_019535 MUS SH3-DOMAIN
GRB2-LIKE 2 MUSCULUS 383956 XM_916921 MUS SIMILAR TO
MAP/MICROTUBULE MUSCULUS AFFINITY-REGULATING KINASE 3 383956
XM_357348 MUS SIMILAR TO MAP/MICROTUBULE MUSCULUS
AFFINITY-REGULATING KINASE 3 245068 XM_918167 MUS SIMILAR TO
MAP/MICROTUBULE MUSCULUS AFFINITY-REGULATING KINASE 4
(MAP/MICROTUBU . . . 245068 XM_142402 MUS SIMILAR TO
MAP/MICROTUBULE MUSCULUS AFFINITY-REGULATING KINASE 4
(MAP/MICROTUBU . . . 238564 NM_001166030 MUS SIMILAR TO MYOSIN
LIGHT CHAIN XM_910560 MUSCULUS KINASE 238564 XM_111421 MUS SIMILAR
TO MYOSIN LIGHT CHAIN MUSCULUS KINASE 229879 XM_143595 MUS GM4862
PREDICTED GENE 4862 XM_001478899 MUSCULUS SIMILAR TO NON-METASTATIC
CELLS 2, PROTEIN (NM23B) EXPRESSED IN ISOFOR . . . GM5374 PREDICTED
GENE 5374 385049 XM_904204 MUS SIMILAR TO RHO-ASSOCIATED
XM_001473528 MUSCULUS COILED-COIL FORMING KINASE 1 385049 XM_358017
MUS SIMILAR TO RHO-ASSOCIATED MUSCULUS COILED-COIL FORMING KINASE 1
17691 NM_010831 MUS SNF1-LIKE KINASE MUSCULUS 74718 NM_029068 MUS
SORTING NEXIN 16 MUSCULUS 67804 NM_026386 MUS SORTING NEXIN 2
MUSCULUS 54198 NM_017472 MUS SORTING NEXIN 3 MUSCULUS 27263
XM_889037 MUS SPERM MOTILITY KINASE 2 MUSCULUS 27263 XM_620264 MUS
SPERM MOTILITY KINASE 2 MUSCULUS 27263 XM_135656 MUS SPERM MOTILITY
KINASE 2 MUSCULUS 27263 XM_889060 MUS SPERM MOTILITY KINASE 2
MUSCULUS 27263 XM_620265 MUS SPERM MOTILITY KINASE 2 MUSCULUS 27263
XM_907097 MUS SPERM MOTILITY KINASE 2 MUSCULUS 20975 NM_011523 MUS
SYNAPTOJANIN 2 MUSCULUS 55925 NM_018802 MUS SYNAPTOTAGMIN VIII
MUSCULUS 319508 NM_176931 MUS SYNAPTOTAGMIN XV MUSCULUS
319508 NM_181529 MUS SYNAPTOTAGMIN XV MUSCULUS 230661 NM_146151 MUS
TESTIS-SPECIFIC KINASE 2 MUSCULUS 58864 NM_080442 MUS
TESTIS-SPECIFIC SERINE KINASE 3 MUSCULUS 56496 NM_019656 MUS
TETRASPANIN 6 MUSCULUS 21912 NM_019634 MUS TETRASPANIN 7 MUSCULUS
21873 NM_011597 MUS TIGHT JUNCTION PROTEIN 2 MUSCULUS 225997
NM_153417 MUS TRANSIENT RECEPTOR POTENTIAL MUSCULUS CATION CHANNEL,
SUBFAMILY M, MEMBER 6 381406 NM_023815 MUS TRP53 REGULATING KINASE
MUSCULUS 76367 NM_023815 MUS TRP53 REGULATING KINASE MUSCULUS 76367
NM_001007581 MUS TRP53 REGULATING KINASE MUSCULUS 381406
NM_001007581 MUS TRP53 REGULATING KINASE MUSCULUS 66745 NM_025741
MUS TUMOR PROTEIN D52-LIKE 3 MUSCULUS 22165 NM_013698 MUS TXK
TYROSINE KINASE MUSCULUS 21846 NM_011587 MUS TYROSINE KINASE
RECEPTOR 1 MUSCULUS 21846 NM_011587 MUS SP. TYROSINE KINASE
RECEPTOR 1 22241 NM_009469 MUS UNC-51 LIKE KINASE 1 (C. ELEGANS)
MUSCULUS 22245 NM_011675 MUS URIDINE-CYTIDINE KINASE 1 MUSCULUS
12929 NM_007764 MUS V-CRK SARCOMA VIRUS CT10 MUSCULUS ONCOGENE
HOMOLOG (AVIAN)-LIKE 11836 NM_009703 MUS V-RAF MURINE SARCOMA 3611
VIRAL MUSCULUS ONCOGENE HOMOLOG 70160 NM_027338 MUS VACUOLAR
PROTEIN SORTING 36 MUSCULUS (YEAST) 20479 NM_009190 MUS VACUOLAR
PROTEIN SORTING 4B MUSCULUS (YEAST) 269338 NM_178851 MUS VPS39
MUSCULUS 269338 NM_147153 MUS VPS39 MUSCULUS 69847 NM_175638 MUS
WNK LYSINE DEFICIENT PROTEIN MUSCULUS KINASE 4 74254 NM_133756 MUS
XPA BINDING PROTEIN 1 MUSCULUS 77976 NM_001004363 MUS ZNUAK FAMILY,
SNF1-LIKE KINASE, 1 MUSCULUS 229309 N/A MUS SIMILAR TO
PHOSPHOGLYCERATE MUSCULUS KINASE (EC 2.7.2.3) - MOUSE 243968 N/A
MUS SIMILAR TO KIAA1883 PROTEIN MUSCULUS 245619 NM_027067 MUS
GLYCOGEN SYNTHASE KINASE 3 [Replaced MUSCULUS ALPHA PROBABLE
PSEUDOGENE with 69389] 268321 N/A MUS SIMILAR TO NUCLEOSIDE
[Replaced MUSCULUS DIPHOSPHATE KINASE B with 432482] 381061
NM_013741 MUS SPERM MOTILITY KINASE 2A [Replaced MUSCULUS with
27263] 381446 N/A MUS SIMILAR TO PHOSPHOGLYCERATE MUSCULUS KINASE
(EC 2.7.2.3) - MOUSE 381981 N/A MUS SIMILAR TO AARF DOMAIN MUSCULUS
CONTAINING KINASE 4 384257 N/A MUS SIMILAR TO NUCLEOSIDE MUSCULUS
DIPHOSPHATE KINASE B 384894 N/A MUS SIMILAR TO CYTOSOLIC THYMIDINE
MUSCULUS KINASE 432447 N/A MUS PHOSPHATIDYLETHANOLAMINE- MUSCULUS
BINDING PROTEIN PSEUDOGENE
[0090] In Tables 1 and 2, the numbers in the column labeled GeneID
correspond with the accession numbers in the Entrez GeneID database
made available by the National Center for Biotechnology Information
(NCBI). The Entrez GeneID may be used to identify corresponding
sequences such as, for example, genomic DNA, mRNA and protein
sequences.
[0091] In Tables 3 and 4, each GeneID is presented along with its
corresponding accession number(s) from the NCBI Reference Sequences
(RefSeq) database for mRNA transcripts. Through the accession
numbers, the sequences are readily available. Table 3 contains
sequences from Table 1. Table 4 contains sequences from Table 2.
The rank order of the GenelDs in Tables 1 and 2 are not maintained
in Tables 3 and 4.
[0092] The terms and expressions which have been employed are used
as terms of descriptions and not of limitation, and there is no
intention that in the use of such terms and expressions of
excluding any equivalents of the features shown and described or
portions thereof, but it is recognized that various modifications
are possible within the scope of the invention. Thus, it should be
understood that although the present invention has been illustrated
by specific embodiments and optional features, modification and/or
variation of the concepts herein disclosed may be resorted to by
those skilled in the art, and that such modifications and
variations are considered to be within the scope of this
invention.
[0093] In addition, where features or aspects of the invention are
described in terms of Markush group or other grouping of
alternatives, those skilled in the art will recognize that the
invention is also thereby described in terms of any individual
member or subgroup of members of the Markush group or other
group.
[0094] All references, including the disclosures of each patent,
patent application, publication and accession number to database
sequences, cited or described in this document are hereby
incorporated herein by reference, in their entireties.
* * * * *