Growth arrest homebox gene

Abstract

A novel growth arrest homeobox gene has been discovered and the nucleotide sequences have been determined in both the rat and the human. The expression of the novel homeobox gene inhibits vascular smooth muscle cell growth. The growth arrest homeobox gene hereinafter referred to as the “Gax gene” and its corresponding proteins are useful in the study of vascular smooth muscle cell proliferation and in the treatment of blood vessel diseases that result from excessive smooth muscle cell proliferation, particularly after balloon angioplasty.

1. An isolated DNA molecule comprising a nucleotide sequence that encodes a mammalian Gax protein, said nucleotide sequence selected from the group consisting of a nucleotide sequence encoding a protein having the amino acid sequence of SEQ ID NO:2 and a nucleotide sequence encoding a protein having the amino acid sequence of SEQ ID NO:4. 
     
     
       2. The DNA molecule of  claim 1 , comprising the nucleotide sequence as shown in ( FIG. 1 ) Sequence Identifier No.  SEQ ID NO: 1. 
     
     
       3. The DNA molecule of  claim 1 , comprising the nucleotide sequence as shown in ( FIG. 3 ) Sequence Identifier No.  SEQ ID NO: 3. 
     
     
       4. The DNA molecule of  claim 1 , comprising the nucleotide sequence encoding a protein having an amino acid sequence as shown in Sequence Identifier No.  SEQ ID NO: 2. 
     
     
       5. The DNA molecule of  claim 1 , comprising a nucleotide sequence having a region which consists of the nucleotide bases from about 749 to about 919 as shown in Sequence Identifier No.  SEQ ID NO: 1. 
     
     
       6. The  An isolated messenger RNA transcript of the DNA of  claim 1 . 
     
     
       7. A vector containing the DNA mol  molecule of  claim 1 . 
     
     
       8. A vector containing the DNA molecule of  claim 2 . 
     
     
       9. A vector containing the DNA molecule of  claim 3 . 
     
     
       10. A vector containing the DNA molecule of  claim 4 . 
     
     
       11. A vector containing the DNA molecule of  claim 5 . 
     
     
       12. A host cell transformed by the vector of  claim 7  containing a DNA molecule having the nucleotide sequence coding for Gax protein. 
     
     
       13. A host cell transformed by the vector of  claim 8  containing a DNA molecule having the nucleotide sequence coding for Gax protein. 
     
     
       14. A host cell transformed by the vector of  claim 9  containing a DNA molecule having the nucleotide sequence coding for Gax protein. 
     
     
       15. A host cell transformed by the vector of  claim 10  containing a DNA molecule having the nucleotide sequence coding for Gax protein. 
     
     
       16. A host cell transformed by the vector of  claim 11  containing a DNA molecule having the nucleotide sequence coding for Gax protein. 
     
     
       17. A process for the preparation of Gax protein comprising culturing the transformed host of  claim 12  under conditions suitable for the expression of Gax protein and recovering the Gax protein. 
     
     
       18. A process for the preparation of a protein which inhibits the proliferation of vascular smooth muscle cells comprising culturing the transformed host of  claim 13  under conditions suitable for the expression of the protein and recovering the protein. 
     
     
       19. A process for the preparation of a protein which inhibits the proliferation of vascular smooth muscle cells comprising culturing the transformed host of  claim 14  under conditions suitable for the expression of the protein and recovering the protein. 
     
     
       20. A process for the preparation of a protein which inhibits the proliferation of vascular smooth muscle cells comprising culturing the transformed host of  claim 15  under conditions suitable for the expression of the protein and recovering the protein. 
     
     
       21. The DNA molecule of  claim 1 , comprising the  a nucleotide sequence encoding a protein having an amino acid sequence as shown in Sequence Identifier No.  SEQ ID NO: 4. 
     
     
       22. A vector containing the DNA molecule of  claim 21 . 
     
     
       23. A host cell transformed by the vector of  claim 22  containing a DNA molecule having the nucleotide sequence encoding the Gax protein. 
     
     
       24. A process for the preparation of a protein which inhibits the proliferation of vascular smooth muscle cell comprising culturing the transformed host of  claim 23  under conditions suitable for the expression of the protein and recovering the protein. 
     
     
       25. The isolated DNA molecule of  claim 1  wherein the protein is a human Gax protein. 
     
     
       26. The isolated DNA molecule of  claim 1  wherein the protein is a rat Gax protein. 
     
     
       27. An isolated DNA molecule comprising a nucleotide sequence encoding a human Gax protein that inhibits proliferation of vascular smooth muscle cells, said nucleotide sequence being 941 nucleotides in length, wherein said nucleotide sequence starting at the 3′ end comprises the following fragments:
 (a) a fragment of clone 6 comprising nucleotides 699 to 941 of said nucleotide sequence;  
 (b) a fragment of clone 23 comprising nucleotides 231 to 698 of said nucleotide sequence;  
 (c) a fragment of clone 117 comprising nucleotides 119 to 230 of said nucleotide sequence;  
 (d) a fragment of clone 131 comprising nucleotides 1 to 118 of said nucleotide sequence.  
 
     
     
       28. The isolated DNA molecule of  claim 1 , comprising a nucleotide sequence having a region which consists of nucleotide  749  to nucleotide  931  of SEQ ID NO: 1 . 
     
     
       29. A process for preparing a host cell for producing Gax protein comprising
 ( a )  introducing the vector according to    claim 7    into a host cell; and      ( b )  culturing the host cell of step  ( a )  under conditions suitable to achieve expression of the DNA molecule contained in said vector.     
     
     
       30. A process for preparing a host cell for producing Gax protein comprising
 ( a )  introducing the vector according to    claim 8    into a host cell; and      ( b )  culturing the host cell of step  ( a )  under conditions suitable to achieve expression of the DNA molecule contained in said vector.     
     
     
       31. A process for preparing a host cell for producing Gax protein comprising
 ( a )  introducing the vector according to    claim 9    into a host cell; and      ( b )  culturing the host cell of step  ( a )  under conditions suitable to achieve expression of the DNA molecule contained in said vector.