Function of cis-acting elements in human alcohol dehydrogenase 4 (ADH4) promoter and role of C/EBP proteins in gene expression.
The ADH4 gene, which encodes human pi-alcohol dehydrogenase, is expressed in a tissue-specific manner, with the highest level in liver and lower levels in the gastrointestinal tract. We examined the location and function of the cis-acting elements that regulate ADH4 transcription. Liver contains proteins that bound to seven sites in the proximal promoter (from bp -387 to bp +17). Proteins from other tissues bound to subsets of these sites and to two additional sites, one of which is a negative cis-acting element. Members of two important transcription factor families, C/EBP and AP-1, bound to several sites in this promoter. The proximal ADH4 promoter functioned in a hepatoma cell line (H4IIE-C3) and a kidney cell line (CV-1). Coexpression of members of the C/EBP family strongly enhanced promoter activity, which can in part explain the high level of expression of ADH4 in liver. At one site that can be bound by both C/EBP and c-Jun, a mutation that abolished binding by C/EBP but not by c-Jun decreased promoter activity in both cell lines. This mutation had a stronger effect in the context of a longer promoter, suggesting interaction among cis-acting elements.
Li
M
M
Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis 46202-5122, USA.
Edenberg
H J
HJ
D000426:Alcohol Dehydrogenase / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D022762:CCAAT-Enhancer-Binding Proteins; D006528:Carcinoma, Hepatocellular; D002457:Cell Extracts; D002460:Cell Line; D002522:Chlorocebus aethiops; D004247:DNA / Q000378:metabolism; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics* / Q000502:physiology; D006801:Humans; D007668:Kidney; D008099:Liver / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics; D014407:Tumor Cells, Cultured
D022762:CCAAT-Enhancer-Binding Proteins; D002457:Cell Extracts; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D016755:Proto-Oncogene Proteins c-jun; D018808:Transcription Factor AP-1; D004247:DNA; D000426:Alcohol Dehydrogenase