Ethanol-induced translocation of cAMP-dependent protein kinase to the nucleus. Mechanism and functional consequences.
Ethanol induces translocation of the catalytic subunit (Calpha) of cAMP-dependent protein kinase (PKA) from the Golgi area to the nucleus in NG108-15 cells. Ethanol also induces translocation of the RIIbeta regulatory subunit of PKA to the nucleus; RI and Cbeta are not translocated. Nuclear PKA activity in ethanol-treated cells is no longer regulated by cAMP. Gel filtration and immunoprecipitation analysis confirm that ethanol blocks the reassociation of Calpha with RII but does not induce dissociation of these subunits. Ethanol also reduces inhibition of Calpha by the PKA inhibitor PKI. Pre-incubation of Calpha with ethanol decreases phosphorylation of Leu-Arg-Arg-Ala-Ser-Leu-Gly (Kemptide) and casein but has no effect on the phosphorylation of highly charged molecules such as histone H1 or protamine. cAMP-response element-binding protein (CREB) phosphorylation by Calpha is also increased in ethanol-treated cells. This increase in CREB phosphorylation is inhibited by the PKA antagonist (R(p))-cAMPS and by an adenosine receptor antagonist. These results suggest that ethanol affects a cascade of events allowing for sustained nuclear localization of Calpha and prolonged CREB phosphorylation. These events may account for ethanol-induced changes in cAMP-dependent gene expression.
Constantinescu
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Department of Neurology, Ernest Gallo Clinic and Research Center, University of California, San Francisco, California 94110-3518, USA.
Diamond
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Gordon
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AS
D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004789:Enzyme Activation; D000431:Ethanol / Q000494:pharmacology*; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D018047:Receptors, Purinergic P1 / Q000378:metabolism; D014407:Tumor Cells, Cultured
D017362:Cyclic AMP Response Element-Binding Protein; D010455:Peptides; D018047:Receptors, Purinergic P1; C061479:protein kinase inhibitor peptide; D000431:Ethanol; D017868:Cyclic AMP-Dependent Protein Kinases