Identification of a FOXA-dependent enhancer of human alcohol dehydrogenase 4 (ADH4)

Sirisha Pochareddy, Howard Edenberg

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Human alcohol dehydrogenase 4 (ADH4) is one of the key enzymes involved in the metabolism of alcohol. ADH4 is highly expressed in the liver, and previous studies have revealed several cis-acting elements in the proximal promoter region. In this study we have identified a distal upstream enhancer, 4E, of ADH4. In HepG2 human hepatoma cells, 4E increased the activity of an ADH4 basal promoter by 50-fold. 4E was cell-specific, as no enhancer activity was detected in a human lung cell line, H1299. We have narrowed the enhancer activity to a 565 bp region and have identified multiple liver-enriched transcription factor binding sites in the region. By electrophoretic mobility shift assays, we confirmed binding of FOXA proteins to these sites. Site-directed mutagenesis studies demonstrated that sites 1 and 4 have the biggest effect on enhancer function, and mutations in multiple sites have multiplicative effects. We also studied the effects of three variations in the minimal enhancer region. Two variations had a significant effect on enhancer activity, decreasing the activity to 0.6-fold, while one had small but significant effect. The differences in the functional activity in different haplotypes suggest that this region could play an important role in the risk for alcoholism.

Original languageEnglish
Pages (from-to)1-7
Number of pages7
JournalGene
Volume460
Issue number1-2
DOIs
StatePublished - Jul 2010

Fingerprint

Liver
Electrophoretic Mobility Shift Assay
Site-Directed Mutagenesis
Genetic Promoter Regions
Haplotypes
Alcoholism
Hepatocellular Carcinoma
Carrier Proteins
Transcription Factors
Binding Sites
Alcohols
Cell Line
Lung
Mutation
alcohol dehydrogenase IV
Enzymes

Keywords

  • Alcoholism
  • Gene regulation
  • Haplotype
  • Polymorphisms
  • Transcription factor

ASJC Scopus subject areas

  • Genetics

Cite this

Identification of a FOXA-dependent enhancer of human alcohol dehydrogenase 4 (ADH4). / Pochareddy, Sirisha; Edenberg, Howard.

In: Gene, Vol. 460, No. 1-2, 07.2010, p. 1-7.

Research output: Contribution to journalArticle

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