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Papers In Press, published online ahead of print September 3, 2002
Nutrition Center and Pediatrics, Arkansas Children's Nutrition Center and University of Arkansas for Mediacl Sciences, Little Rock, AR 72202
Corresponding Author: badgerthomasm{at}uams.edu
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/EBP
J. Biol. Chem, 10.1074/jbc.M204535200
Submitted on May 8, 2002
Revised on July 19, 2002
Accepted on September 3, 2002
Ethanol induction of class I alcohol dehydrogenase expression in the rat occurs through induction of CCAAT/Enhancer binding proteins
and 
to the C/EBP site. Western blot immunoblot analysis of HNEs demonstrated 3.5- and 2.3-fold increases in C/EBP (LAP) and C/EBP
(p<0.05), respectively, in ethanol-fed rats compared with controls, while levels of the truncated C/EBP (LIP) and C/EBP
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. Anti-sera against LIP or C/EBP 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 and .
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