Human preprotachykinin gene promoter

Abstract

Compositions and methods are provided for identifying novel therapeutic agents for the treatment of breast cancer, bone marrow metastasis, pain, arthritis, aggressive behavior, depression, and certain hematopoietic disorders. Disclosed are promoters and 3′ regulatory regions of genes whose expression differs in malignant cells as compared with non-malignant cells. These include PPT-I, NK-2 and SP-R.

We claim:  
     
         1 . An isolated nucleic acid molecule comprising a PPT-I gene promoter region, corresponding to a segment of a human PPT-I gene commencing at about 722 nucleotides upstream of a transcription start site of the PPT-I gene, and encompassing the transcription start site of the gene.  
     
     
         2 . The nucleic acid molecule of  claim 1 , wherein said nucleic acid molecule comprises a one or more cAMP response elements.  
     
     
         3 . The nucleic acid molecule of  claim 1 , isolated from a PPT-I gene that hybridizes under moderate conditions to a coding sequence comprising SEQ ID NO:15.  
     
     
         4 . The nucleic acid molecule of  claim 1 , comprising a sequence contained in SEQ ID NO:1 or SEQ ID NO:14.  
     
     
         5 . An expression vector comprising the nucleic acid molecule of  claim 1 , operably linked to a heterologous gene that encodes a gene product.  
     
     
         6 . The vector of  claim 5 , wherein the heterologous gene is a reporter gene.  
     
     
         7 . The vector of  claim 6 , wherein the gene is selected from the group consisting of luciferase, secreted alkaline phosphatase, GFP and chloramphenicol acetyltransferase.  
     
     
         8 . An isolated host cell transformed with the vector of  claim 5 .  
     
     
         9 . The host cell of  claim 8  which is a mammalian cell.  
     
     
         10 . The host cell of  claim 9  which is a human cell.  
     
     
         11 . A nucleic acid molecule comprising the sequence shown in SEQ ID NO:1, or a fragment of said sequence shown in SEQ ID NO:1, said fragment comprising at least about 500 nucleotides, said nucleic acid sequence exhibiting promoter activity.  
     
     
         12 . An expression vector comprising the nucleic acid molecule of  claim 11 .  
     
     
         13 . A method for determining the presence of an alteration in a promoter region of a PPT-I gene, the alteration being associated with a pathological condition, the method comprising: 
 a) providing a nucleic acid molecule of SEQ ID NO:1;    b) isolating the corresponding sequence from a test subject suspected of having an alteration in the PPT-I promoter;    c) forming a heteroduplex between the sequence of step and step b); and    d) assessing the heteroduplex for the presence of an alteration selected from the group consisting of a mismatch, an insertion and a deletion.    
     
     
         14 . An isolated nucleic acid molecule comprising a PPT-I gene 3′ region, isolated from a PPT-I gene that hybridizes under moderate conditions to a coding sequence comprising SEQ ID NO:15.  
     
     
         15 . The nucleic acid molecule of  claim 14 , comprising a sequence contained in SEQ ID NO:4.  
     
     
         16 . An expression vector comprising the nucleic acid molecule of  claim 14 , operably linked to a heterologous gene that encodes a gene product.  
     
     
         17 . The vector of  claim 16 , wherein the heterologous gene is a reporter gene.  
     
     
         18 . The vector of  claim 16 , wherein the gene is selected from the group consisting of luciferase, secreted alkaline phosphatase, GFP and chloramphenicol acetyltransferase.  
     
     
         19 . An isolated host cell transformed with the vector of  claim 16 .  
     
     
         20 . The host cell of  claim 19  which is a mammalian cell.  
     
     
         21 . The host cell of  claim 19  which is a human cell.  
     
     
         22 . A nucleic acid molecule comprising the sequence shown in SEQ ID NO:4, or a fragment of said sequence shown in SEQ ID NO:4, said fragment comprising at least about 200 nucleotides, said nucleic acid sequence exhibiting expression regulatory activity.  
     
     
         23 . An expression vector comprising the nucleic acid molecule of  claim 22 .  
     
     
         24 . A method for determining the presence of an alteration in the 3′ region of a PPT-I gene, said alteration being associated with a pathological condition, said method comprising: 
 a) providing a nucleic acid molecule of SEQ ID NO:4;  
 b) isolating the corresponding sequence from a test subject suspected of having an alteration in the 3′ region of PPT-I;  
 c) forming a heteroduplex between the sequence of step and step b); and  
 d) assessing the heteroduplex for the presence of an alteration selected from the group consisting of a mismatch, an insertion and a deletion.