title: Identification of cyclic AMP response element in the human renin gene.
abstract: In order to identify the mechanism by which cyclic AMP stimulates expression of the human renin gene (REN), the effect of forskolin was tested in transient expression analyses of REN 5'-flanking DNA-chloramphenicol acetyltransferase (CAT) reporter gene constructs in secondary cultures of human chorio-decidual cells, a major site of renin synthesis. Forskolin induced a mean 5-fold stimulation which was localized to DNA in the region -249 to -162 with respect to the transcription start site (+1). Such DNA also mediated a response to forskolin in heterologous (HSV thymidine kinase) promoter constructs. Strong cAMP-response element (CRE) homology at -222 to -218 resembled the target for members of the CRE binding protein (CREB) family. Gel shift assays demonstrated similarly migrating nucleoprotein complexes for oligonucleotides containing the putative REN CRE as for a canonical CRE, in chorio-decidual, JEG-3 and HeLa nuclear extracts. Mutation of residues critical for CREB attachment reduced binding. In conclusion, a CRE was identified at -222 to -218 that appears critical for cAMP-induced human renin gene transcription.
authors.affiliation: Department of Medicine, University of Southern California Medical Center, Los Angeles 90033.
authors.forename: D L
authors.initials: DL
authors.lastname: Smith
authors.forename: B J
authors.initials: BJ
authors.lastname: Morris
authors.forename: Y S
authors.initials: YS
authors.lastname: Do
authors.forename: R E
authors.initials: RE
authors.lastname: Law
authors.forename: K J
authors.initials: KJ
authors.lastname: Shaw
authors.forename: W A
authors.initials: WA
authors.lastname: Hseuh
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D015186:Hominidae / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D010920:Placenta / Q000201:enzymology; D011247:Pregnancy; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000502:physiology*; D012083:Renin / Q000096:biosynthesis / Q000235:genetics*; D015183:Restriction Mapping; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000096:biosynthesis; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D009838:Oligodeoxyribonucleotides; D005576:Colforsin; D004247:DNA; D000242:Cyclic AMP; D013937:Thymidine Kinase; D012083:Renin