title: Lysine acetylation induced by chronic ethanol consumption impairs dynamin-mediated clathrin-coated vesicle release.
abstract: The liver is the major site of ethanol metabolism and thus sustains the most injury from chronic alcohol consumption. Ethanol metabolism by the hepatocyte leads to the generation of reactive metabolites and oxygen radicals that can readily adduct DNA, lipids, and proteins. More recently, it has become apparent that ethanol consumption also leads to increased post-translational modifications of the natural repertoire, including lysine hyperacetylation. Previously, we determined that alcohol consumption selectively impairs clathrin-mediated internalization in polarized hepatocytes. However, neither the step at which the block occurs nor the mechanism responsible for the defect have been identified. To identify the specific step at which clathrin-mediated internalization is impaired, we examined the distributions, levels, and assembly of selected components of the clathrin machinery in control and ethanol-treated cells. To determine whether the impairment is caused by ethanol-induced lysine acetylation, we also examined the same coat components in cells treated with trichostatin A (TSA), a deacetylase inhibitor that leads to protein hyperacetylation in the absence of ethanol.
  
  
  CONCLUSIONS
  We determined that both ethanol and TSA impair internalization at a late stage before vesicle fission. We further determined that this defect is likely the result of decreased dynamin recruitment to the necks of clathrin-coated invaginations resulting in impaired vesicle budding. These results also raise the exciting possibility that agents that promote lysine deacetylation may be effective therapeutics for the treatment of alcoholic liver disease.
authors.affiliation: Department of Biology, The Catholic University of America, Washington, DC 20064, USA.
authors.forename: Blythe D
authors.initials: BD
authors.lastname: Shepard
authors.forename: Dean J
authors.initials: DJ
authors.lastname: Tuma
authors.forename: Pamela L
authors.initials: PL
authors.lastname: Tuma
mesh_terms: D000107:Acetylation / Q000187:drug effects; D000428:Alcohol Drinking / Q000378:metabolism*; D000818:Animals; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D022163:Clathrin-Coated Vesicles / Q000187:drug effects / Q000378:metabolism*; D034281:Dynamins / Q000187:drug effects / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology*; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006877:Hydroxamic Acids / Q000494:pharmacology; D008239:Lysine / Q000378:metabolism*; D023421:Models, Animal; D051381:Rats
chemical_list: D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D000431:Ethanol; C012589:trichostatin A; D034281:Dynamins; D008239:Lysine
references.citation: Alcohol Clin Exp Res. 1997 Sep;21(6):1073-82
references.pmid: 9309320
references.citation: Dev Cell. 2006 Jun;10(6):839-50
references.pmid: 16740485
references.citation: J Cell Biol. 1994 Nov;127(4):915-34
references.pmid: 7962076
references.citation: Alcohol Res Health. 2003;27(4):285-90
references.pmid: 15540799
references.citation: Am J Physiol. 1998 Apr;274(4):G757-66
references.pmid: 9575859
references.citation: Biochem Pharmacol. 2004 Jun 1;67(11):2167-74
references.pmid: 15135311
references.citation: J Cell Biol. 1983 Aug;97(2):499-507
references.pmid: 6411734
references.citation: J Cell Biol. 1993 Dec;123(6 Pt 2):1761-75
references.pmid: 7506266
references.citation: J Cell Biol. 1987 Sep;105(3):1241-51
references.pmid: 3654750
references.citation: Mol Cell. 2007 Jul 20;27(2):197-213
references.pmid: 17643370
references.citation: J Hepatol. 2006 May;44(5):963-70
references.pmid: 16169115
references.citation: Biochem Pharmacol. 2009 Sep 15;78(6):648-55
references.pmid: 19463792
references.citation: Proc Natl Acad Sci U S A. 1989 Jul;86(13):4968-72
references.pmid: 2472637
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Hepatology. 2002 Dec;36(6):1478-87
references.pmid: 12447874
references.citation: Alcohol Clin Exp Res. 1987 Dec;11(6):579-84
references.pmid: 3124658
references.citation: Alcohol. 1999 Nov;19(3):255-60
references.pmid: 10580516
references.citation: Life Sci. 1989;45(16):1461-6
references.pmid: 2509834
references.citation: Alcohol Clin Exp Res. 2010 Feb;34(2):280-91
references.pmid: 19951295
references.citation: Hepatology. 2008 Nov;48(5):1671-9
references.pmid: 18697214
references.citation: World J Gastroenterol. 2009 Mar 14;15(10):1219-30
references.pmid: 19291822
references.citation: J Biol Chem. 1994 Dec 9;269(49):30842-7
references.pmid: 7983015
references.citation: Mol Biol Cell. 2002 Jan;13(1):225-37
references.pmid: 11809835
references.citation: Alcohol Clin Exp Res. 2006 Nov;30(11):1906-15
references.pmid: 17067356
references.citation: Science. 2006 Sep 15;313(5793):1591-4
references.pmid: 16973870
references.citation: Biochem Pharmacol. 1996 Nov 22;52(10):1499-505
references.pmid: 8937463
references.citation: J Cell Sci. 1994 Apr;107 ( Pt 4):813-25
references.pmid: 8056838
references.citation: Hepatology. 2008 May;47(5):1745-53
references.pmid: 18161881
references.citation: Semin Liver Dis. 1988 Feb;8(1):69-80
references.pmid: 3283943
references.citation: Alcohol Alcohol Suppl. 1991;1:297-303
references.pmid: 1669008
references.citation: Cell. 1983 May;33(1):273-85
references.pmid: 6147196
references.citation: Nature. 2003 Mar 6;422(6927):37-44
references.pmid: 12621426
references.citation: Am J Physiol Gastrointest Liver Physiol. 2008 Oct;295(4):G833-42
references.pmid: 18755807
references.citation: Curr Opin Investig Drugs. 2008 Apr;9(4):371-8
references.pmid: 18393104