Characterization, sequence, and expression of the cloned human neuropeptide Y gene.

The human neuropeptide Y (NPY) gene was isolated from a human genomic DNA library. The transcription unit spans approximately 8 kilobase pairs and is interrupted by three intervening sequences. The first exon contains only nontranslated DNA. The site where transcription initiates was determined by primer extension analysis using a primer derived from a human cDNA, pheochromocytoma RNA and avian myeloblastosis virus reverse transcriptase. A TATA-like sequence and a CAAT-like sequence occur 25 and 70 base pairs 5' to the transcription start site, respectively. The second exon begins with the initiator Met for preproNPY and extends to the Arg (residue 63) which precedes the Tyr-amide of mature NPY. The third exon contains the coding region for 27 amino acids, and the fourth exon codes for the terminal heptapeptide and the 3' nontranslated DNA. Transcriptional control elements were investigated by fusing 581 base pairs of the 5' sequences of the NPY gene to the promoterless structural gene for chloramphenicol acetyltransferase. NPY promoter activity was assayed by transfection of these hybrid constructions into CA-77 and PC12 cells followed by the determination of chloramphenicol acetyltransferase activity in cellular extracts. DNA sequences located within 530 bases of the start of transcription are sufficient for transient expression in the two neuronally derived cell lines examined.