Mechanism of action of a repressor of dioxin-dependent induction of Cyp1a1 gene transcription

A. J. Watson, K. I. Weir-Brown, R. M. Bannister, F. F. Chu, S. Reisz-Porszasz, Y. Fujii-Kuriyama, K. Sogawa, O. Hankinson

    Research output: Contribution to journalArticlepeer-review

    62 Citations (Scopus)


    A dominant mutant of Hepa-1 cells, c31, expresses a repressor that prevents 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-dependent stimulation of Cyp1a1 transcription. The repressor acts via the xenobiotic-responsive elements (XREs), which are the DNA-binding sites for the aryl hydrocarbon (Ah) receptor-TCDD complex during transcriptional activation of the gene. High-salt nuclear extracts prepared from c31 cells grown with TCDD contained normal levels of the Ah receptor which bound the XRE with normal affinity, as judged by in vitro gel mobility shift assays. Furthermore, extracts prepared from these cells, grown either with or without TCDD, contained no novel XRE- binding proteins compared with extracts from wild-type Hepa-1 cells. However, in vivo genomic footprinting demonstrated that TCDD treatment leads to binding of the Ah receptor to the XREs in Hepa-1 but not mutant cells. This finding suggests that the repressor associates with the Ah receptor to prevent its binding to the XREs and that high-salt treatment either causes dissociation of the receptor/repressor complex or fails to extract the repressor from nuclei. The results underscore the importance of using both in vivo and in vitro assays for analyzing DNA-protein interactions.

    Original languageEnglish
    Pages (from-to)2115-2123
    Number of pages9
    JournalMolecular and cellular biology
    Issue number5
    Publication statusPublished - 1992

    ASJC Scopus subject areas

    • Molecular Biology
    • Cell Biology

    Fingerprint Dive into the research topics of 'Mechanism of action of a repressor of dioxin-dependent induction of Cyp1a1 gene transcription'. Together they form a unique fingerprint.

    Cite this