The adult enhancer factor-1, a Drosophila melanogaster transcriptional repressor, modulates the promoter activity of the rat class-I alcohol dehydrogenase-encoding gene

Gene. 1994 Nov 18;149(2):325-30. doi: 10.1016/0378-1119(94)90170-8.

Abstract

Expression of the Drosophila melanogaster alcohol dehydrogenase-encoding gene (ADH) in the adult fat body is controlled by the ADH adult enhancer site (AAE). The D. melanogaster transcription repressor, adult enhancer factor-1 (AEF-1), binds to AAE at a site which overlaps with a sequence recognized by the mammalian transcription factor, CCAAT/enhancer-binding protein alpha [C/EBP alpha; Falb and Maniatis, Genes Dev. 6 (1992a) 454-465]. C/EBP alpha also activates the promoter of the rat class-I ADH gene in a sequence-specific manner [Potter et al., Arch. Biochem. Biophys. 285 (1991a) 246-251]. In this study, we explored the possibility that D. melanogaster AEF-1 influences transcription of the rat class-I ADH. By DNase I footprint analysis, bacterially produced AEF-1 protects a region of DNA between nucleotides (nt) -22 and -36 of the rat class-I ADH promoter (pADH), just 5' to the binding site of C/EBP alpha, a result confirmed by the electrophoretic mobility shift assay (EMSA). Co-transfection of a rat pADH-CAT reporter construct with expression vectors containing C/EBP alpha, AEF-1, or both, indicates that AEF-1 inhibits induction of the rat pADH by C/EBP alpha. Moreover, rat liver nuclear extracts appear to contain AEF-1-like-binding activities to AAE by EMSA. These experiments suggest an evolutionarily conserved mechanism by which AEF-1 modulates expression of the D. melanogaster and rat ADH genes.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alcohol Dehydrogenase / biosynthesis
  • Alcohol Dehydrogenase / genetics*
  • Animals
  • Base Sequence
  • Binding Sites
  • CCAAT-Enhancer-Binding Proteins
  • DNA / metabolism
  • DNA-Binding Proteins / metabolism*
  • Deoxyribonuclease I
  • Drosophila Proteins
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / metabolism*
  • Gene Expression
  • Isoenzymes / biosynthesis
  • Isoenzymes / genetics
  • Molecular Sequence Data
  • Mutagenesis
  • Nuclear Proteins / metabolism*
  • Oligodeoxyribonucleotides / chemistry
  • Oligodeoxyribonucleotides / metabolism
  • Promoter Regions, Genetic*
  • Rats
  • Repressor Proteins / metabolism*
  • Sequence Homology, Nucleic Acid
  • Transcription Factors / metabolism*
  • Transfection

Substances

  • Aef1 protein, Drosophila
  • CCAAT-Enhancer-Binding Proteins
  • DNA-Binding Proteins
  • Drosophila Proteins
  • Isoenzymes
  • Nuclear Proteins
  • Oligodeoxyribonucleotides
  • Repressor Proteins
  • Transcription Factors
  • DNA
  • Alcohol Dehydrogenase
  • Deoxyribonuclease I