Ethanol Alters the Subcellular Localization of δ- and ε Protein Kinase C in NG108–15 Cells
- 1Ernest Gallo Clinic and Research Center and Department of Neurology (A.S.G., L.Y., Z.-L.W., I.R.C., I.D.), 2Department of Cellular and Molecular Pharmacology and Neuroscience Program (A.S.G., I.D.), University of California, San Francisco, California 94110
Abstract
Protein kinase C (PKC) has been shown to regulate the ethanol sensitivity of membrane-bound receptors and transporters, but little is known about the molecular mechanisms underlying this regulation. PKC is a family of isozymes that translocate to new intracellular sites on activation. Here we present immunochemical data showing that ethanol causes translocation of δ- and ε-PKC to new intracellular sites. Ethanol causes translocation of δ-PKC from the Golgi to the perinucleus; this translocation is similar to that induced by activation of PKC with phorbol esters. In contrast, ε-PKC translocation caused by ethanol is different from that induced by phorbol esters; ethanol causes translocation of ε-PKC from the perinucleus to the cytoplasm, whereas phorbol ester activation causes translocation of ε-PKC to the nucleus. Because the substrate specificity of these kinases is determined by their site of localization, ethanol-induced translocation of δ- and ε-PKC to new intracellular sites may explain some of the pleiotropic effects of ethanol on cellular functions.
Footnotes
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Send reprint requests to: Adrienne S. Gordon, Ph.D., Ernest Gallo Clinic and Research Center, San Francisco General Hospital, 1001 Potrero Avenue, Building 1, #101, San Francisco, CA 94110-3518. E-mail:adrienn{at}itsa.ucsf.edu
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↵1 Current affiliation: Exelixis Pharmaceuticals, Oakland, CA 94606.
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↵2 Current affiliation: Department of Biology, York University, North York, Ontario M3J 1P3, Canada.
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↵3 Wu, Z.-L, unpublished observations.
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This work was supported in part by National Institutes of Health Grant AA10039.
- Abbreviations:
- PKC
- protein kinase C
- HEPES
- 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- PBS
- phosphate-buffered saline
- EGTA
- ethylene glycol bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid
- TBS
- Tris-buffered saline
- PMA
- phorbol 12-myristate 13-acetate
- DAG
- diacylglycerol
- RACK
- receptor for activated C kinase
- PKA
- cAMP-dependent protein kinase
- ANOVA
- analysis of variance
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- Received June 10, 1997.
- Accepted July 10, 1997.
- The American Society for Pharmacology and Experimental Therapeutics
