Myosin IXa and cyclic nucleotide gated channel-15 (CNGC-15) polynucleotides, polypeptides, compositions, methods, and uses thereof

Abstract

The present invention discloses the amino acid and nucleic acid sequences of a new CNGC and Myosin that map to the region of the human chromosome associated with Bardet-Biedl Syndrome. Cyclic nucleotide gated channels (CNGCs) comprise a family of multimeric protein ion channels that open in response to the binding of a cyclic nucleotide to an intracellular domain. The two new proteins, CNGC-15 and Myosin IXa, are useful in the study, diagnosis and treatment of Bardet-Biedl Syndrome and Usher Syndrome. Other indications that can be treated by CNGC-15 and/or Myosin IXa polypeptides, or agonists or antagonists include hearing loss, retinis pigmentosa, obesity, hypogonadism, sterility, polydactyly, brachydactyly, syndactyly, mental retardation, renal abnormalities, hypertension, diabetes and cardiovascular abnormalities. Compositions and methods for expressing cyclic nucleotide gated channel-15 (CNGC-15) and Myosin IXa are provided. The compositions comprise CNGC-15 and Myosin IXa polypeptides and derivatives thereof, nucleotide sequences, expression cassettes, transformed cells and antibodies to these polypeptides. Methods for the expression and detection of CNGC-15 and Myosin IXa nucleotides and polypeptides and compositions for the treatment of these conditions are provided.

That which is claimed:  
     
         1 . A native Myosin IXa polypeptide substantially free of other human proteins.  
     
     
         2 . An active fragment or derivative of a Myosin IXa polypeptide.  
     
     
         3 . A Myosin IXa polypeptide comprising an amino acid sequence selected from the group consisting of: 
 a) SEQ IDNO: 1;    b) residues 1-146 of SEQ ID NO: 1;    c) residues 147-719 of SEQ ID NO: 1;    d) residues 239-246 of SEQ ID NO: 1;    e) residues 720-835 of SEQ ID NO: 1;    f) residues 872-1166 of SEQ ID NO: 1;    g) residues 2074-2219 of SEQ ID NO: 1;    h) a polypeptide having at least about 61% homology to SEQ ID NO: 1;    i) a polypeptide at least about 95% homology to an amino acid sequence of b), c), d), e), f) or g); and    j) a polypeptide of at least about 90 contiguous residues of SEQ ID NO: 1.    
     
     
         4 . A fusion protein comprising the Myosin IXa polypeptide of  claim 3 , or fragment thereof, and a polypeptide of interest.  
     
     
         5 . A native CNGC-15 polypeptide substantially free of other human proteins.  
     
     
         6 . An active fragment or derivative of a CNGC-15 polypeptide.  
     
     
         7 . A CNGC-15 polypeptide comprising an amino acid sequence selected from the group consisting of: 
 a) a polypeptide encoded by SEQ ID NO: 3;    b) a polypeptide encoded by nucleotides 369-1307 of SEQ ID NO: 3;    c) a polypeptide having at least about 80% homology to the polypeptide encoded by nucleotides 369-1307 of SEQ ID NO: 3; and    d) a polypeptide of at least about 20 contiguous residues of the polypeptide encoded by nucleotides 369-1307 of SEQ ID NO: 3.    
     
     
         8 . A fusion protein comprising the CNGC-15 polypeptide of  claim 7 , or fragment thereof, and a polypeptide of interest.  
     
     
         9 . A nucleotide sequence encoding the fusion protein of  claim 4 .  
     
     
         10 . A nucleotide sequence encoding the fusion protein of  claim 8 .  
     
     
         11 . An isolated nucleotide sequence selected from the group consisting of: 
 a) a nucleotide sequence encoding a native Myosin IXa polypeptide;    b) a nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 1;    c) the nucleotide sequence of SEQ ID NO: 2;    d) a nucleotide sequence comprising at least about 390 contiguous bases of SEQ ID NO: 2;    e) a nucleotide sequence comprising nucleotides 243-7085 of SEQ ID NO: 2;    f) a nucleotide sequence comprising nucleotides 243-680 of SEQ ID NO: 2;    g) a nucleotide sequence comprising nucleotides 683-2399 of SEQ ID NO: 2;    h) a nucleotide sequence comprising nucleotides 959-977 of SEQ ID NO: 2;    i) a nucleotide sequence comprising nucleotides 2402-2747 of SEQ ID NO: 2;    j) a nucleotide sequence comprising nucleotides 3158-3740 of SEQ ID NO: 2; and    k) a nucleotide sequence comprising nucleotides 6473-6899 of SEQ ID NO: 2;    
     
     
         12 . An isolated nucleotide sequence selected from the group consisting of: 
 a) a nucleotide sequence encoding a native CNGC-15 polypeptide;    b) a nucleotide sequence encoding the polypeptide encoded by nucleotides 369-1307 of SEQ ID NO: 3;    c) the nucleotide sequence of SEQ ID NO: 3;    d) a nucleotide sequence comprising at least 45 contiguous nucleotides of the sequence of SEQ ID NO: 3; and    e) a nucleotide sequence that hybridizes under stringent conditions to a denatured DNA having a nucleotide sequence of a), b), c) or d).    
     
     
         13 . An expression vector comprising the nucleotide sequence of  claim 9  wherein said nucleotide sequence is operably linked to a promoter.  
     
     
         14 . An expression vector comprising the nucleotide sequence of  claim 10  wherein said nucleotide sequence is operably linked to a promoter.  
     
     
         15 . An expression vector comprising the nucleotide sequence of  claim 11  wherein said nucleotide sequence is operably linked to a promoter.  
     
     
         16 . An expression vector comprising the nucleotide sequence of  claim 12  wherein said nucleotide sequence is operably linked to a promoter.  
     
     
         17 . A transformed cell having stably incorporated into its genome the expression vector of  claim 13 .  
     
     
         18 . A transformed cell having stably incorporated into its genome the expression vector of  claim 14 .  
     
     
         19 . A transformed cell having stably incorporated into its genome the expression vector of  claim 15 .  
     
     
         20 . A transformed cell having stably incorporated into its genome the expression vector of  claim 16 .  
     
     
         21 . A method for producing a Myosin IXa polypeptide or a derivative thereof, comprising: 
 a) providing a host cell stably transformed with an expression vector comprising a promoter operably linked to a nucleotide sequence encoding said Myosin IXa polypeptide or derivative thereof; and    b) culturing the host cell under conditions which allow expression of said polypeptide.    
     
     
         22 . A method for producing a CNGC-15 polypeptide or a derivative thereof, comprising: 
 a) providing a host cell stably transformed with an expression vector comprising a promoter operably linked to a nucleotide sequence encoding said CNGC-15 polypeptide or derivative thereof; and    b) culturing the host cell under conditions which allow expression of said polypeptide.    
     
     
         23 . A method for detecting an unconventional Myosin polynucleotide comprising: 
 a) providing a nucleic acid probe which hybridizes a nucleotide sequence of claim  11 ;    b) contacting the probe with a sample of polynucleotides under hybridizing conditions to form a duplex; and    c) detecting said duplexes.    
     
     
         24 . A method for detecting a CNGC polynucleotide comprising: 
 a) providing a nucleic acid probe which hybridizes a nucleotide sequence of claim  12 ;    b) contacting the probe with a sample of polynucleotides under hybridizing conditions to form a duplex; and    c) detecting said duplexes.    
     
     
         25 . An antibody specific for a CNGC-15 or Myosin IXa polypeptide.  
     
     
         26 . A method for detecting a CNGC-15 polypeptide comprising: 
 a) providing an antibody that binds to a CNGC-15 polypeptide;    b) contacting the antibody to a sample under binding conditions to form a duplex; and    c) detecting said duplexes.    
     
     
         27 . A method for detecting a Myosin IXa polypeptide comprising: 
 a) providing an antibody that binds to a Myosin IXa polypeptide;    b) contacting the antibody to a sample under binding conditions to form a duplex; and    c) detecting said duplexes.    
     
     
         28 . A method for treating Bardet-Biedl Syndrome, Usher Syndrome, hearing loss, retinis pigmentosa, obesity, hypogonadism, sterility, polydactyly, brachydactyly, syndactyly, mental retardation, renal abnormalities, hypertension, diabetes and cardiovascular abnormalities, comprising administering a therapeutically effective amount of a CNGC-15 polypeptide to a subject in need of such treatment.  
     
     
         29 . A method for treating Bardet-Biedl Syndrome, Usher Syndrome, hearing loss, retinis pigmentosa, obesity, hypogonadism, sterility, polydactyly, brachydactyly, syndactyly, mental retardation, renal abnormalities, hypertension, diabetes and cardiovascular abnormalities, comprising administering a therapeutically effective amount of a Myosin-IXA polypeptide to a subject in need of such treatment.  
     
     
         30 . A composition comprising a CNGC-15 polypeptide and a pharmaceutically acceptable carrier.  
     
     
         31 . A composition comprising a Myosin IXa polypeptide and a pharmaceutically acceptable carrier.