Protein-DNA interactions in the 5' region of the mouse alcohol dehydrogenase gene Adh-1.
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 in 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.
Department of Biochemistry, Indiana University School of Medicine, Indianapolis 46223.
L G
LG
Carr
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Zhang
H J
HJ
Edenberg
D000426:Alcohol Dehydrogenase / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008745:Methylation; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D015345:Oligonucleotide Probes; D004247:DNA; D000426:Alcohol Dehydrogenase; D015500:Chloramphenicol O-Acetyltransferase