Protein-DNA interactions in the 5′ region of the mouse alcohol dehydrogenase gene Adh-1

Lucinda G. Carr, Zhang Ke, Howard J. Edenberg

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

The mouse alcohol dehydrogenase-encoding gene, Adh-1, is expressed at high levels in adult liver. We have begun analysis of the regulation of this gene, focusing upon specific DNA-protein interactions. Preliminary deletion mapping of the 5′ region indicated that a 521-bp fragment extending from nucleotide (nt) -468 to + 53 (relative to the transcription start point) could direct chloramphenicol acetyl transferase synthesis m hepatoma cells. We therefore focused upon this -468 to + 53 fragment. Using the gel mobility-shift assay, we detected at least four different complexes between proteins extracted from nuclei of mouse liver or hepatoma cells and regions within the -468 to + 53 fragment. Two of the DNA-protein complexes can be competed with a 43-bp region from nt -90 to -48, and an oligodeoxyribonucleotide spanning this region forms two complexes. The strongest of these two DNA-protein complexes has been localized by methylation interference experiments to the palindromic sequence CACGTG located between nt -57 and -62. This region is identical in the related human ADH2 gene, and may represent a novel regulatory sequence.

Original languageEnglish (US)
Pages (from-to)277-285
Number of pages9
JournalGene
Volume78
Issue number2
DOIs
StatePublished - May 30 1989

Keywords

  • cis-acting sequences
  • gel retardation
  • Gene expression
  • liver
  • methyl interference
  • nuclear proteins

ASJC Scopus subject areas

  • Genetics

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