title: Angiotensinogen gene expression is stimulated by the cAMP-responsive element binding protein in opossum kidney cells.
abstract: It has been reported previously that the addition of isoproterenol or forskolin stimulates the expression of the angiotensinogen (ANG) gene in opossum kidney (OK) 27 cells, an OK cell line with a fusion gene containing the 5'-flanking regulatory sequence of the rat ANG gene fused with a human growth hormone (hGH) gene as a reporter, pOGH (ANG N-1498/+18), permanently integrated into their genomes. To investigate whether the effect of isoproterenol or forskolin on the expression of the ANG gene is mediated via the nuclear 43-kD cAMP-responsive element binding protein (CREB), OK 27 cells were transiently transfected with an expression plasmid containing the cDNA for the 43-kD CREB (pRSV/CREB). The level of expression of the pOGH (ANG N-1498/+18) in OK 27 cells was estimated by the amount of immunoreactive hGH secreted into the culture medium. Transfection of pRSV/CREB alone stimulated the expression of pOGH (ANG N-1498/+18). The addition of isoproterenol or forskolin further enhanced the stimulatory effect of pRSV/ CREB on the expression of pOGH (ANG N-1498/+18). The enhancing effect of isoproterenol was inhibited by the presence of propranolol (an inhibitor of beta-adrenoceptors) and (R)-p-adenosine 3'5'-cyclic monophospho-orthioate (Rp)-cAMP (an inhibitor of cAMP-dependent protein kinase A I and II). Transfection of pRSV/CREB had no effect on the expression of thymidine kinase growth hormone in OK 13 cells, an OK cell line with a fusion gene containing the promoter/enhancer DNA sequence of the viral thymidine-kinase gene fused with an hGH gene as a reporter, thymidine kinase growth hormone, permanently integrated into their genomes. These studies demonstrate that isoproterenol stimulates the expression of ANG gene via the cAMP-dependent protein kinase A and probably via the interaction of the 43-kD CREB with the 5'-flanking region of the ANG gene. Our data indicate that the nuclear 43-kD CREB may have a modulatory role on the expression of the ANG gene in OK cells.
authors.affiliation: University of Montreal, Maisonneuve-Rosemont Hospital, Research Center, Quebec, Canada.
authors.forename: J F
authors.initials: JF
authors.lastname: Qian
authors.forename: T T
authors.initials: TT
authors.lastname: Wang
authors.forename: X H
authors.initials: XH
authors.lastname: Wu
authors.forename: J
authors.initials: J
authors.lastname: Wu
authors.forename: C
authors.initials: C
authors.lastname: Ge
authors.forename: S
authors.initials: S
authors.lastname: Lachance
authors.forename: S
authors.initials: S
authors.lastname: Carriere
authors.forename: J S
authors.initials: JS
authors.lastname: Chan
mesh_terms: D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000319:Adrenergic beta-Antagonists / Q000494:pharmacology; D000808:Angiotensinogen / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D019382:Human Growth Hormone / Q000235:genetics; D006801:Humans; D007545:Isoproterenol / Q000494:pharmacology; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D009893:Opossums; D011433:Propranolol / Q000494:pharmacology; D051381:Rats; D013873:Thionucleotides / Q000494:pharmacology; D014162:Transfection
chemical_list: D000318:Adrenergic beta-Agonists; D000319:Adrenergic beta-Antagonists; D017362:Cyclic AMP Response Element-Binding Protein; D017931:DNA Primers; D018076:DNA, Complementary; D013873:Thionucleotides; D000808:Angiotensinogen; D019382:Human Growth Hormone; D005576:Colforsin; C016957:adenosine-3',5'-cyclic phosphorothioate; D011433:Propranolol; D000242:Cyclic AMP; D007545:Isoproterenol