Cloning and characterization of the 5 '-flanking region of the human prolactin-releasing peptide receptor gene

Recently a novel peptide which specifically stimulates the secretion of pro lactin (PRL) was found and named PRL-releasing peptide (PrRP). To evaluate the regulation of human (h) PrRP-receptor (PrRP-R) gene expression, we clon ed the 5'-flanking region of the hPrRP-R gene and determined the nucleotide sequence of 4.0 kilobase pairs (kb) upstream from the translation start si te. Analysis of the hPrRP-R transcripts by means of 5'-rapid amplification of cDNA ends suggested that the hPrRP-R gene had multiple transcription sta rt sites between -429 and -365 from the translation start site. There is no typical TATA or CAAT but a GC box and putative binding sites for several t ranscription factors including Pit-1 and pituitary homeobox 1 (Ptx1), Howev er, transient transfection studies using a luciferase reporter gene demonst rated that 5'-flanking region exerts promoter activity in several non-pitui tary cell lines as well as in GH(3) cells. The GC box located from -467 to -457 was identified as an important region for the basal expression of the hPrRP-R gene. Knowledge of the promoter region of the hPrRP-R gene, which w as obtained in the present study, will facilitate the clarification of its transcriptional regulation. (C) 2000 Academic Press.