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Originally published In Press as doi:10.1074/jbc.M204535200 on September 3, 2002

J. Biol. Chem., Vol. 277, Issue 46, 43572-43577, November 15, 2002
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Ethanol Induction of Class I Alcohol Dehydrogenase Expression in the Rat Occurs through Alterations in CCAAT/Enhancer Binding Proteins beta  and gamma *

Ling He, Martin J. J. Ronis, and Thomas M. BadgerDagger

From the Arkansas Children's Nutrition Center, Little Rock, Arkansas 72202

Alcohol dehydrogenase (ADH) is the principal ethanol-metabolizing enzyme. Ethanol induces rat Class I ADH mRNA and activity by an as yet unknown mechanism. In the current study, adult male rats were fed an ethanol-containing diet by continuous intragastric infusion for 42 days. Hepatic Class I ADH mRNA, protein, and activity levels in the ethanol-infused rats increased 3.9-, 3.3-, and 1.7-fold, respectively (p <0.05). Cis-acting elements within the proximal promoter region of the ADH gene were studied by electrophoretic mobility shift assay (EMSA). Hepatic nuclear extract (HNE) binding to either the consensus or ADH-specific CCAAT/enhancer binding protein (C/EBP) sites was >2.4-fold greater in ethanol-fed rats (p <0.05) than controls. Antibody-specific EMSA assays demonstrated binding of the transcription factor C/EBPbeta to the C/EBP site. Western blot immunoblot analysis of HNEs demonstrated 3.5- and 2.3-fold increases in C/EBPbeta (LAP) and C/EBPdelta (p <0.05), respectively, in ethanol-fed rats compared with controls, whereas levels of the truncated C/EBPbeta (LIP) and C/EBPgamma were lower in ethanol-fed rats (p <0.05). HNE from ethanol-fed rats increased (3-fold) the in vitro transcription of rat Class I ADH (p <0.05), and mutation of the C/EBP element in the proximal promoter region blocked this effect. Antisera against LIP or C/EBPgamma enhanced transcription efficiency (p <0.05). These data provide the first evidence for the mechanism by which ethanol regulates rat hepatic Class I ADH gene expression in vivo. This mechanism involves the C/EBP site and the enhancer binding proteins beta  and gamma .

* This work was supported in part by National Institutes of Health Grant AA0845.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Arkansas Children's Nutrition Center, 1120 Marshall St., Little Rock, AR 72202. Tel.: 501-364-2785; Fax: 501-364-2818; E-mail: badgerthomasm@uams.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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