Premium
An inverted cAMP response element mediates the cAMP induction of the ovine β1‐adrenergic receptor gene
Author(s) -
Tseng Yi T.,
Stabila Joan,
McGonnigal Beth,
Nguyen Tien T.,
Padbury James F.
Publication year - 1998
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549800204682
Subject(s) - creb , microbiology and biotechnology , cyclic amp response element binding protein , protein kinase a , transfection , gene , response element , biology , protein subunit , luciferase , creb1 , binding site , gene expression , chemistry , promoter , transcription factor , kinase , biochemistry
We identified an inverted, functional cAMP response element (CRE) located at ‐1599 bp relative to the translation start site within the ovine β1‐adrenergic receptor (β1AR) gene promoter. In transfection studies with SK‐N‐MC cells, a 40‐bp oligonucleotide containing the potential CRE, β1AR‐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 α‐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 β1AR‐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 β1AR gene expression by interacting with an inverted CRE within the promoter region. This is the first reported functional CRE among all β1AR genes.