32142, 21481, 25964, 21686, novel human dehydrogenase molecules and uses thereof

Abstract

The invention provides isolated nucleic acids molecules, designated DHDR nucleic acid molecules, which encode novel DHDR-related dehydrogenase molecules. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing DHDR nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a DHDR gene has been introduced or disrupted. The invention still further provides isolated DHDR proteins, fusion proteins, antigenic peptides and anti-DHDR antibodies. Diagnostic methods utilizing compositions of the invention are also provided.

What is claimed:  
     
         1 . An isolated nucleic acid molecule selected from the group consisting of: 
 (a) a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, or SEQ ID NO:10; and    (b) a nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9 or SEQ ID NO:12.    
     
     
         2 . An isolated nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:11.  
     
     
         3 . An isolated nucleic acid molecule comprising the nucleotide sequence contained in the plasmid deposited with ATCC® as Accession Number ______.  
     
     
         4 . An isolated nucleic acid molecule which encodes a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:11.  
     
     
         5 . An isolated nucleic acid molecule selected from the group consisting of: 
 a) a nucleic acid molecule comprising a nucleotide sequence which is at least 60% identical to the nucleotide sequence of SEQ ID NO:1 or 3, SEQ ID NO: 4 or 6, SEQ ID NO:7 or 9, or SEQ ID NO: 10 or 12, or a complement thereof;    b) a nucleic acid molecule comprising a fragment of at least 50 nucleotides of a nucleic acid comprising the nucleotide sequence of SEQ ID NO:1 or 3, SEQ ID NO: 4 or 6, SEQ ID NO:7 or 9, or SEQ ID NO:10 or 12, or a complement thereof;    c) a nucleic acid molecule which encodes a polypeptide comprising an amino acid sequence at least about 60% identical to the amino acid sequence of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:12; and    d) a nucleic acid molecule which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:11, wherein the fragment comprises at least 16 contiguous amino acid residues of the amino acid sequence of SEQ ID NO: 2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:11.    
     
     
         6 . An isolated nucleic acid molecule which hybridizes to the nucleic acid molecule of any one of claims  1 ,  2 ,  3 ,  4 , or  5  under stringent conditions.  
     
     
         7 . An isolated nucleic acid molecule comprising a nucleotide sequence which is complementary to the nucleotide sequence of the nucleic acid molecule of any one of claims  1 ,  2 ,  3 ,  4 , or  5 .  
     
     
         8 . An isolated nucleic acid molecule comprising the nucleic acid molecule of any one of claims  1 ,  2 ,  3 ,  4 , or  5 , and a nucleotide sequence encoding a heterologous polypeptide.  
     
     
         9 . A vector comprising the nucleic acid molecule of any one of claims  1 ,  2 ,  3 ,  4 , or  5 .  
     
     
         10 . The vector of  claim 9 , which is an expression vector.  
     
     
         11 . A host cell transfected with the expression vector of  claim 10 .  
     
     
         12 . A method of producing a polypeptide comprising culturing the host cell of  claim 11  in an appropriate culture medium to, thereby, produce the polypeptide.  
     
     
         13 . An isolated polypeptide selected from the group consisting of: 
 a) a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:11, wherein the fragment comprises at least 16 contiguous amino acids of SEQ ID NO: 2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:11;    b) a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:11, wherein the polypeptide is encoded by a nucleic acid molecule which hybridizes to a nucleic acid molecule consisting of SEQ ID NO:1 or 3, SEQ ID NO: 4 or 6, SEQ ID NO:7 or 9, or SEQ ID NO:10 or 12 under stringent conditions;    c) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 60% identical to a nucleic acid comprising the nucleotide sequence of SEQ ID NO:1, 3, 4, 6, 7, 9, 10, or 12;    d) a polypeptide comprising an amino acid sequence which is at least 60% identical to the amino acid sequence of SEQ ID NO: 2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:11.    
     
     
         14 . The isolated polypeptide of  claim 13  comprising the amino acid sequence of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:11.  
     
     
         15 . The polypeptide of  claim 13 , further comprising heterologous amino acid sequences.  
     
     
         16 . An antibody which selectively binds to a polypeptide of  claim 13 .  
     
     
         17 . A method for detecting the presence of a polypeptide of  claim 13  in a sample comprising: 
 a) contacting the sample with a compound which selectively binds to the polypeptide; and  
 b) determining whether the compound binds to the polypeptide in the sample to thereby detect the presence of a polypeptide of  claim 13  in the sample.  
 
     
     
         18 . The method of  claim 17 , wherein the compound which binds to the polypeptide is an antibody.  
     
     
         19 . A kit comprising a compound which selectively binds to a polypeptide of  claim 13  and instructions for use.  
     
     
         20 . A method for detecting the presence of a nucleic acid molecule of any one of claims  1 ,  2 ,  3 ,  4 , or  5  in a sample comprising: 
 a) contacting the sample with a nucleic acid probe or primer which selectively hybridizes to the nucleic acid molecule; and  
 b) determining whether the nucleic acid probe or primer binds to a nucleic acid molecule in the sample to thereby detect the presence of a nucleic acid molecule of any one of claims  1 ,  2 ,  3 ,  4 , or  5  in the sample.  
 
     
     
         21 . The method of  claim 20 , wherein the sample comprises mRNA molecules and is contacted with a nucleic acid probe.  
     
     
         22 . A kit comprising a compound which selectively hybridizes to a nucleic acid molecule of any one of claims  1 ,  2 ,  3 ,  4 , or  5  and instructions for use.  
     
     
         23 . A method for identifying a compound which binds to a polypeptide of  claim 13  comprising: 
 a) contacting the polypeptide, or a cell expressing the polypeptide with a test compound; and  
 b) determining whether the polypeptide binds to the test compound.  
 
     
     
         24 . The method of  claim 23 , wherein the binding of the test compound to the polypeptide is detected by a method selected from the group consisting of: 
 a) detection of binding by direct detection of test compound/polypeptide binding;    b) detection of binding using a competition binding assay; and    c) detection of binding using an assay for DHDR activity.    
     
     
         25 . A method for modulating the activity of a polypeptide of  claim 13  comprising contacting the polypeptide or a cell expressing the polypeptide with a compound which binds to the polypeptide in a sufficient concentration to modulate the activity of the polypeptide.  
     
     
         26 . A method for identifying a compound which modulates the activity of a polypeptide of  claim 13  comprising: 
 a) contacting a polypeptide of  claim 13  with a test compound; and  
 b) determining the effect of the test compound on the activity of the polypeptide to thereby identify a compound which modulates the activity of the polypeptide.  
 
     
     
         27 . A method of identifying a subject having a viral disorder, or at risk for developing a viral disorder comprising: 
 a) contacting a sample obtained from said subject comprising nucleic acid molecules with a hybridization probe comprising at least 25 contiguous nucleotides of SEQ ID NO:10; and    b) detecting the presence of a nucleic acid molecule in said sample that hybridizes to said probe, thereby identifying a subject having a viral disorder, or at risk for developing a viral disorder.    
     
     
         28 . The method of  claim 27 , wherein said hybridization probe is detectably labeled.  
     
     
         29 . The method of  claim 27 , wherein said sample comprising nucleic acid molecules is subjected to agarose gel electrophoresis and southern blotting prior to contacting with said hybridization probe.  
     
     
         30 . The method of  claim 29 , wherein said method is used to detect genomic DNA in said sample.  
     
     
         31 . The method of  claim 27 , wherein said sample comprising nucleic acid molecules is subjected to agarose gel electrophoresis and northern blotting prior to contacting with said hybridization probe.  
     
     
         32 . The method of  claim 31 , wherein said method is used to detect mRNA in the sample.  
     
     
         33 . The method of  claim 27 , wherein said detecting is by in situ hybridization.  
     
     
         34 . A method of identifying a subject having a viral disorder, or at risk for developing a viral disorder comprising: 
 a) contacting a sample obtained from said subject comprising nucleic acid molecules with a first and a second amplification primer, said first primer comprising at least 25 contiguous nucleotides of SEQ ID NO:10 and said second primer comprising at least 25 contiguous nucleotides from the complement of SEQ ID NO:10;    b) incubating said sample under conditions that allow nucleic acid amplification; and    c) detecting the presence of a nucleic acid molecule in said sample that is amplified, thereby identifying a subject having a viral disorder, or at risk for developing a viral disorder.    
     
     
         35 . The method of  claim 34 , wherein said sample comprising nucleic acid molecules is subjected to agarose gel electrophoresis after said incubation step.  
     
     
         36 . The method of any one of claims  34 , wherein said method is used to detect mRNA in said sample.  
     
     
         37 . The method of any one of claims  34 , wherein said method is used to detect genomic DNA in said sample.  
     
     
         38 . A method of identifying a subject having a viral disorder, or at risk for developing a viral disorder comprising: 
 a) contacting a sample obtained from said subject comprising polypeptides with a DHDR binding substance; and    b) detecting the presence of a polypeptide in said sample that binds to said DHDR binding substance, thereby identifying a subject having a viral disorder or at risk for developing a viral disorder.    
     
     
         39 . The method of  claim 38 , wherein said binding substance is an antibody.  
     
     
         40 . The method of  claim 38 , wherein said binding substance is detectably labeled.  
     
     
         41 . A method for identifying a compound capable of treating a viral disorder characterized by aberrant DHDR nucleic acid expression or DHDR polypeptide activity comprising assaying the ability of the compound to modulate DHDR nucleic acid expression or DHDR polypeptide activity, thereby identifying a compound capable of treating a viral disorder characterized by aberrant DHDR nucleic acid expression or DHDR polypeptide activity.  
     
     
         42 . The method of  claim 41 , wherein the disorder is associated with hepatitis B virus infection.  
     
     
         43 . A method for treating a subject having a viral disorder characterized by aberrant DHDR polypeptide activity or aberrant DHDR nucleic acid expression comprising administering to the subject a DHDR modulator, thereby treating said subject having a viral disorder.  
     
     
         44 . The method of  claim 43 , wherein the DHDR modulator is a small molecule.  
     
     
         45 . The method of  claim 43 , wherein the DHDR modulator is an antisense oligonucleotide.  
     
     
         46 . The method of  claim 43 , wherein the DHDR modulator is a ribozyme.  
     
     
         47 . The method of  claim 43 , wherein the DHDR modulator is a polypeptide.  
     
     
         48 . The method of  claim 43 , wherein the DHDR modulator is an antibody.  
     
     
         49 . The method of  claim 43 , wherein the disorder is associated with hepatitis B virus infection.