The cAMP response element binding protein is expressed and phosphorylated in cardiac myocytes.
Cardiac cells grow in response to a number of stimuli that activate intracellular signaling pathways. The cAMP-signaling pathway mediates the activation of gene transcription in other cell types by the cAMP response element binding protein (CREB-P). Our aim was to explore the physiological role of CREB-P in response to elevated cAMP in cardiac cells by determining if phosphorylation of CREB-P (to phosphoCREB-P) rapidly induces transcription in culture. Primary embryonic chick heart cultures were used in which cAMP was raised by forskolin (5 mumol/L) or isoproterenol (10 mumol/L) treatment. Since both these agents have inotropic effects, tension production was controlled with 2,3-butanedione monoxime (BDM). This allowed us to determine whether the cAMP-signaling pathway or the contractile state was regulating phosphorylation and transcription. The responses for time periods up to 2 hours were assayed with antibodies to detect phosphoCREB-P and by quantitative filter hybridization for creb gene expression. The staining intensity of the phosphoprotein increased in myocyte nuclei after 10 minutes and persisted for 1 hour with either forskolin or isoproterenol treatment. An increase in creb mRNA abundance was also detected, with the maximum level of expression being at 1 hour with forskolin treatment. These changes are independent of the contractile state, because BDM itself caused no change. BDM plus forskolin induced the same pattern of creb expression as observed with forskolin alone. Therefore, we conclude that elevation of cAMP leads to phosphorylation of CREB-P and an increase in creb mRNA abundance.
Goldspink
P H
PH
Department of Physiology and Biophysics, College of Medicine, University of Illinois at Chicago 60612-7342.
Russell
B
B
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000096:biosynthesis; D017362:Cyclic AMP Response Element-Binding Protein / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D003931:Diacetyl / Q000031:analogs & derivatives / Q000494:pharmacology; D015870:Gene Expression; D009206:Myocardium / Q000378:metabolism*; D010766:Phosphorylation
D017362:Cyclic AMP Response Element-Binding Protein; C004717:diacetylmonoxime; D005576:Colforsin; D000242:Cyclic AMP; D003931:Diacetyl