Specific protein-DNA interactions at a xenobiotic-responsive element: Copurification of dioxin receptor and DNA-binding activity

J. Hapgood, S. Cuthill, M. Denis, L. Poellinger, J. A. Gustafsson

Research output: Contribution to journalArticle

76 Scopus citations

Abstract

Upon binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin (called dioxin or TCDD), the dioxin receptor exhibits increased affinity for the cell nucleus in vivo and for DNA in vitro. To define the recognition sequence of the dioxin receptor and its relationship with that of the glucocorticoid receptor, oligonucleotides derived from dioxin-responsive elements of the rat cytochrome P-450c gene were tested for their ability to form specific protein-DNA complexes in a gel retardation assay. We found that a previously defined sequence motif that is similar to the glucocorticoid-responsive element and exhibits strong enhancer activity in response to dioxin receptor ligands bound a dioxin-inducible factor with high specificity but was not recognized by the DNA-binding domain of the glucocorticoid receptor. Binding to this element was only observed in nuclear extracts of wild-type mouse hepatoma cells in a time- and dose-dependent manner and not in nuclear extracts from a nonresponsive mutant cell line deficient in DNA binding of the dioxin receptor. The specific DNA-binding activity in wild-type nuclear extracts comigrated in a Superose size-exclusion column and cosedimented on sucrose gradients with the in vivo labeled dioxin receptor. These experiments strongly suggest that the dioxin receptor is a sequence-specific DNA-binding protein and is not only biochemically but also functionally similar to the steroid receptor family.

Original languageEnglish (US)
Pages (from-to)60-64
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume86
Issue number1
DOIs
StatePublished - 1989

ASJC Scopus subject areas

  • General

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