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 prolactin (PRL) was found and named PRL-releasing peptide (PrRP). To evaluate the regulation of human (h) PrRP-receptor (PrRP-R) gene expression, we cloned 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 site. Analysis of the hPrRP-R transcripts by means of 5'-rapid amplification of cDNA ends suggested that the hPrRP-R gene had multiple transcription start 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 transcription factors including Pit-1 and pituitary homeobox 1 (Ptx1). However, transient transfection studies using a luciferase reporter gene demonstrated that 5'-flanking region exerts promoter activity in several non-pituitary 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 was obtained in the present study, will facilitate the clarification of its transcriptional regulation.
Kishimoto
M
M
Third Division, Kobe University School of Medicine, Kobe, 650-0017, Japan.
Okimura
Y
Y
Hinuma
S
S
Fukusumi
S
S
Iguchi
G
G
Fumoto
M
M
Iida
K
K
Kaji
H
H
Chihara
K
K
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D005786:Gene Expression Regulation; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051381:Rats; D018013:Receptors, Neuropeptide / Q000235:genetics*
D018013:Receptors, Neuropeptide; C400801:receptor, prolactin-releasing peptide; D004247:DNA