Isoproterenol response following transfection of the mouse beta 2-adrenergic receptor gene into Y1 cells.
The beta 2-adrenergic receptor (beta 2AR) gene was isolated from a mouse genomic library, sequenced and shown to share 93% identity with the hamster beta 2AR cDNA at the amino acid level. This mouse beta 2AR genomic clone was transfected into the Y1 mouse adrenal cortex tumor cell line. Northern blot and S1 nuclease analysis showed that the beta 2AR-transfected cells expressed an mRNA of the appropriate size to encode the receptor. Membrane receptor number and affinities for various beta-adrenergic agonists demonstrated that the transfected clone encoded a beta 2AR protein product. Incubation of the transfected Y1 cells, which do not normally possess beta 2AR, with the beta 2AR agonist, isoproterenol, resulted in an increase in the rate of steroid secretion by these cells as well as a rapid change in cell morphology. This response was fully blocked by the beta 2AR antagonist, propranolol. Prolonged incubation of the cells with isoproterenol resulted in agonist insensitivity and an 80% reduction in membrane receptor number.
Allen
J M
JM
Howard Hughes Medical Institute, Department of Biochemistry, University of Washington, Seattle 98195.
Baetge
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EE
Abrass
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IB
Palmiter
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D005796:Genes; D007545:Isoproterenol / Q000494:pharmacology*; D007700:Kinetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011433:Propranolol / Q000494:pharmacology; D011943:Receptors, Adrenergic, beta / Q000187:drug effects / Q000235:genetics*; D013154:Spleen / Q000378:metabolism; D013256:Steroids / Q000096:biosynthesis / Q000378:metabolism; D014162:Transfection
D011943:Receptors, Adrenergic, beta; D013256:Steroids; D011433:Propranolol; D007545:Isoproterenol