Dioxin- and Ah receptor-dependent protein binding to xenobiotic responsive elements and G-rich DNA studied by in vivo footprinting.
DNA-protein interactions before and after transcriptional activation of the carcinogen- and dioxin-inducible enhancer of the murine CYP1A1 gene were detected in vivo by treatment with dimethyl sulfate followed by ligation-mediated, polymerase chain reaction-aided genomic sequencing. Following 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) treatment of mouse Hepa-1 hepatoma cells, evidence of protein binding was detected at the sequence 5' CACGCNA/T 3' within two previously defined xenobiotic response elements (XREs). The observed XRE footprint was similar to that previously identified by in vitro methylation protection footprints and attributed to the binding protein for 2,3,7,8-tetrachlorodibenzo-p-dioxin the Ah receptor. No XRE footprinting was observed in Hepa-1 mutant cells possessing a defective Ah receptor. Unexpectedly, evidence of protein binding was also detected at a G-rich DNA sequence immediately adjacent to one of the XREs. Footprinting of the G-rich sequence element, like that of XRE1 and XRE2, was dependent on the presence of a functional Ah receptor. The Ah receptor is therefore able to bind to its own DNA target sites in vivo and is also required for the binding of a second factor to the G-rich element.
Watson
A J
AJ
Laboratory of Biomedical and Environmental Sciences, University of California, Los Angeles 90024.
Hankinson
O
O
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000187:drug effects / Q000378:metabolism*; D016172:DNA Fingerprinting; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006147:Guanine / Q000378:metabolism*; D008745:Methylation; D051379:Mice; D008969:Molecular Sequence Data; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity*; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug / Q000378:metabolism*; D015262:Xenobiotics / Q000633:toxicity*
D004268:DNA-Binding Proteins; D000072317:Polychlorinated Dibenzodioxins; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug; D015262:Xenobiotics; D006147:Guanine; D004247:DNA