An inverted cAMP response element mediates the cAMP induction of the ovine beta 1-adrenergic receptor gene.
We identified an inverted, functional cAMP response element (CRE) located at--1599 bp relative to the translation start site within the ovine beta 1-adrenergic receptor (beta 1 AR) gene promoter. In transfection studies with SK-N-MC cells, a 40-bp oligonucleotide containing the potential CRE, beta 1 AR-CRE, conferred a 3- to 4-fold increase in luciferase activity mediated by cAMP. The induction was mimicked by co-transfecting the cells with a vector overexpressing the alpha-catalytic subunit of the cAMP-dependent protein kinase (PKA) without treatment, and was blocked by overexpressing a PKA inhibitor (PKI). In electrophoretic mobility shift assays, a discrete binding pattern was shown in cell nuclear extract probed with the 40 bp beta 1 AR-CRE. The binding was shown to be specific and supershifted by addition of a CRE binding protein (CREB-1) antibody. These data demonstrate that cAMP mediates the induction of beta 1 AR gene expression by interacting with an inverted CRE within the promoter region. This is the first reported functional CRE among all beta 1 AR genes.
Tseng
Y T
YT
Dept. Pediatrics, Women & Infants' Hospital of Rhode Island, Brown University, Providence, USA. Yi-Tang_Tseng@brown.edu
Stabila
J
J
McGonnigal
B
B
Nguyen
T T
TT
Padbury
J F
JF
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003994:Bucladesine / Q000494:pharmacology; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D016384:Consensus Sequence; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D007545:Isoproterenol / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics; D008253:Macaca mulatta; D051379:Mice; D009838:Oligodeoxyribonucleotides; D014176:Protein Biosynthesis; D051381:Rats; D018340:Receptors, Adrenergic, alpha-1 / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D012756:Sheep; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection
D017362:Cyclic AMP Response Element-Binding Protein; D009838:Oligodeoxyribonucleotides; D018340:Receptors, Adrenergic, alpha-1; D011993:Recombinant Fusion Proteins; D005576:Colforsin; D003994:Bucladesine; D000242:Cyclic AMP; D008156:Luciferases; D017868:Cyclic AMP-Dependent Protein Kinases; D007545:Isoproterenol