The binding of transformed aromatic hydrocarbon (Ah) receptor to its DNA recognition site is not affected by metal depletion.
The biological effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin), a potent environmental contaminant, are mediated by a soluble intracellular protein, the aromatic hydrocarbon (Ah) receptor (AhR). TCDD:AhR complexes activate gene transcription by binding to specific DNA sequences termed dioxin-responsive elements adjacent to TCDD-responsive genes. Analogies between the AhR and receptors for steroid hormones imply similarities in their mechanism of action. The presence of chelatable, protein-bound metal(s), presumably zinc, is required for DNA binding of several proteins, including steroid hormone receptors and the transcription factor SP1. Utilizing gel retardation and DNA-cellulose binding assays we have investigated the importance of metal in DNA binding of transformed TCDD:AhR complexes. Here, we report that although 1,10-phenanthroline, a metal ion chelating agent, inhibited the DNA binding of SP1 and transformed glucocorticoid receptor, no inhibition of transformed AhR was observed. EDTA was similarly ineffective in inhibiting DNA binding of transformed AhR. Our findings suggest that the AhR, although similar to steroid receptors, appears not to require metals for binding to its specific DNA recognition sequence.
Denison
M S
MS
Department of Biochemistry, Michigan State University, East Lansing 48823.
Deal
R M
RM
D000818:Animals; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004147:Dioxins / Q000378:metabolism*; D004492:Edetic Acid / Q000494:pharmacology; D008297:Male; D010618:Phenanthrolines / Q000494:pharmacology; D000072317:Polychlorinated Dibenzodioxins / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D016329:Sp1 Transcription Factor; D014157:Transcription Factors / Q000378:metabolism; D015032:Zinc / Q000494:pharmacology*
D004268:DNA-Binding Proteins; D004147:Dioxins; D010618:Phenanthrolines; D000072317:Polychlorinated Dibenzodioxins; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug; D011965:Receptors, Glucocorticoid; D016329:Sp1 Transcription Factor; D014157:Transcription Factors; D004247:DNA; D004492:Edetic Acid; D015032:Zinc