Cloning and characterization of the 5'-flanking region of the human growth hormone secretagogue receptor gene.
Recently, the growth hormone secretagogue receptor (GHS-R) cDNA has been isolated from the pituitary and hypothalamus. To evaluate the regulation of human (h) GHS-R gene expression, we cloned the hGHS-R gene containing the 5'-flanking region of 0.6-2.9 kilobase pairs. Analysis of the hGHS-R transcripts with 5'-rapid amplification of cDNA ends suggested that the putative transcription initiation site was approximately -453 base pairs upstream of the translation initiation site (+1). There is no typical TATA, CAAT, or GC box but an initiator-like sequence and putative binding sites for several transcription factors around the putative transcription start site. The 5'-flanking region inserted into a luciferase reporter vector had promoter activity in GH3 cells but had activity indistinguishable from background in HeLa or EP1 cells. The hGHS-R promoter activity in GH3 cells increased by deletion of nucleotides from -1224 to -734, whereas it was decreased by further deletion from -734 to -608. Knowledge of the promoter region of the hGHS-R gene will facilitate elucidation of its transcriptional control.
Kaji
H
H
Third Division, Department of Medicine, Kobe University School of Medicine, Kobe 650-0017, Japan. hkaji@med.kobe-u.ac.jp
Tai
S
S
Okimura
Y
Y
Iguchi
G
G
Takahashi
Y
Y
Abe
H
H
Chihara
K
K
D001483:Base Sequence; D003001:Cloning, Molecular; D019204:GTP-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D015698:Genomic Library; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010902:Pituitary Gland / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D019105:Random Amplified Polymorphic DNA Technique; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000235:genetics*; D043562:Receptors, G-Protein-Coupled; D054440:Receptors, Ghrelin; D011994:Recombinant Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D017384:Sequence Deletion; D014158:Transcription, Genetic; D014162:Transfection
D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D054440:Receptors, Ghrelin; D011994:Recombinant Proteins; D019204:GTP-Binding Proteins