32263, a novel human biotin enzyme and uses thereof

Abstract

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

1 . An isolated nucleic acid molecule selected from the group consisting of: 
 a) a nucleic acid molecule comprising a nucleotide sequence which is at least 80% identical to the nucleotide sequence of SEQ ID NO:1 or 3;    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;    c) a nucleic acid molecule which which hybridizes to a nucleic acid molecule consisting of SEQ ID NO:1 or 3 under stringent conditions;    d) a nucleic acid molecule which encodes a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, wherein the fragment comprises at least 15 contiguous amino acid residues of the amino acid sequence of SEQ ID NO: 2; and    e) the complement of the nucleic acid molecule of any of a), b), c) or d).    
     
     
         2 . An isolated nucleic acid molecule comprising the nucleic acid molecule of  claim 1  and a nucleotide sequence encoding a heterologous polypeptide.  
     
     
         3 . A vector comprising the nucleic acid molecule of  claim 1 .  
     
     
         4 . A host cell which expresses the nucleic acid molecule of  claim 1 .  
     
     
         5 . A method of producing a polypeptide comprising culturing the host cell of  claim 4  in an appropriate culture medium to, thereby, produce the polypeptide.  
     
     
         6 . An isolated polypeptide selected from the group consisting of: 
 a) a polypeptide comprising a fragment of the amino acid sequence of SEQ ID NO: 2, wherein the fragment comprises at least 15 contiguous amino acids of SEQ ID NO: 2;    b) a polypeptide comprising a variant of the amino acid sequence of SEQ ID NO:2, wherein the variant is encoded by a nucleic acid molecule which hybridizes to a nucleic acid molecule consisting of SEQ ID NO:1 or 3 under stringent conditions;    c) a polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 80% identical to the nucleotide sequence of SEQ ID NO:1 or 3;    d) a polypeptide comprising an amino acid sequence which is at least 80% identical to the amino acid sequence of SEQ ID NO: 2.    
     
     
         7 . The polypeptide of  claim 6 , further comprising heterologous amino acid sequences.  
     
     
         8 . An antibody which selectively binds to the polypeptide of  claim 6 .  
     
     
         9 . A method for detecting the presence of the polypeptide of  claim 6  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 the polypeptide of  claim 6  in the sample.    
     
     
         10 . The method of  claim 9 , wherein the compound which binds to the polypeptide is an antibody.  
     
     
         11 . A kit comprising a compound which selectively binds to the polypeptide of  claim 6  and instructions for use.  
     
     
         12 . A method for detecting the presence of the nucleic acid molecule of  claim 1  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 the nucleic acid molecule in the sample to thereby detect the presence of a nucleic acid molecule of  claim 1  in the sample.    
     
     
         13 . The method of  claim 12 , wherein the sample comprises mRNA molecules and is contacted with a nucleic acid probe.  
     
     
         14 . A kit comprising a compound which selectively hybridizes to the nucleic acid molecule of  claim 1  and instructions for use.  
     
     
         15 . A method for identifying a compound which binds to the polypeptide of  claim 6  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.    
     
     
         16 . The method of  claim 15 , 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 BRE activity.    
     
     
         17 . A method for modulating the activity of the polypeptide of  claim 6  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.  
     
     
         18 . A method for identifying a compound which modulates the activity of the polypeptide of  claim 6  comprising: 
 a) contacting the polypeptide of  claim 6  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.