DNA binding activity of the aryl hydrocarbon receptor is sensitive to redox changes in intact cells.
The potential involvement of vicinal dithiols in the transformation of the aryl hydrocarbon (Ah) receptor from its ligand binding to DNA binding form in Hepa-1 cells was explored through the use of diamide and phenylarsine oxide (PAO), which have been shown to specifically form a stable ring complex with vicinal sulfhydryl groups in selected proteins. Pretreatment with diamide and PAO rapidly prevented the inducer-dependent formation of the Ah receptor/xenobiotic response element complex detected by electrophoretic mobility shift assays and suppressed Ah receptor-mediated transcription. Diamide and PAO also inhibited DNA binding activity of the nuclear Ah receptor subsequent to its translocation to the nucleus but to a lesser extent than that observed with pretreatment conditions. The Ah receptor exhibited much higher sensitivity to cellular redox changes than Sp1, a transcription factor previously shown to be very sensitive to redox regulation. Diamide added to nuclear extracts inhibited Ah receptor DNA binding more than when it was added in intact cells. In contrast, Ah receptor DNA binding activity was more sensitive to PAO when it was added to intact cells than when it was added to nuclear extracts. Finally, dithiol 2,3-dimercaptopropanol was over 100 times more effective than monothiol 2-mercaptoethanol in reversing the PAO-dependent inhibition of Ah receptor DNA binding activity. This suggests that vicinal sulfhydryl residues may be involved in DNA binding of the Ah receptor.
Xu
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C
Department of Pharmacognosy, University of Mississippi, University, Mississippi, 38677, USA.
Siu
C S
CS
Pasco
D S
DS
D000818:Animals; D001152:Arsenicals / Q000494:pharmacology; D001692:Biological Transport; D006528:Carcinoma, Hepatocellular; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis / Q000235:genetics; D004273:DNA, Neoplasm / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D003958:Diamide / Q000494:pharmacology; D004112:Dimercaprol / Q000494:pharmacology; D008623:Mercaptoethanol / Q000494:pharmacology; D051379:Mice; D010084:Oxidation-Reduction; D018336:Receptors, Aryl Hydrocarbon / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016329:Sp1 Transcription Factor / Q000187:drug effects / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured; D015262:Xenobiotics / Q000494:pharmacology
D001152:Arsenicals; D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D018336:Receptors, Aryl Hydrocarbon; D016329:Sp1 Transcription Factor; D015262:Xenobiotics; D004112:Dimercaprol; C029341:oxophenylarsine; D003958:Diamide; D008623:Mercaptoethanol; D019363:Cytochrome P-450 CYP1A1