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Molecular Pharmacology

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Abstract

In vitro transformation of the human Ah receptor and its binding to a dioxin response element.

P A Harper, J V Giannone, A B Okey and M S Denison
Molecular Pharmacology October 1992, 42 (4) 603-612;
P A Harper
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J V Giannone
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A B Okey
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M S Denison
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Abstract

Many biological effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin) are mediated by a soluble intracellular protein, the Ah receptor (AhR). After binding of TCDD to the cytoplasmic AhR there occurs a poorly understood "transformation" step, wherein the TCDD-AhR complex is converted to a form that can bind to DNA with high affinity. The binding of transformed AhR to a specific dioxin-responsive element (DRE) upstream of a given gene stimulates transcriptional activation of that gene. Using a gel retardation assay we examined the interaction of transformed human cytosolic TCDD-AhR complexes with a synthetic DNA oligonucleotide containing a single DRE site. Transformation and DNA binding of human AhR in vitro was ligand dependent and specific for DRE-containing DNA. Unlike rodent hepatic AhR, in vitro transformation of human AhR was completely temperature dependent. Although at 4 degrees AhR binds ligand, no transformation of human TCDD-AhR complex was observed at 4 degrees even after 24 h; however, rapid transformation as measured by DNA binding was detectable as early as 10 min after warming to 22 degrees, with maximal binding by about 60 min. Calf thymus DNA-Sepharose or DRE-Sepharose column chromatography showed that transformed human cytosolic AhR interacts with DNA as a single species. The absolute temperature dependency of human AhR transformation mimics that observed in vivo and provides a useful system to study the mechanism of AhR transformation in detail.

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Molecular Pharmacology
Vol. 42, Issue 4
1 Oct 1992
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Abstract

In vitro transformation of the human Ah receptor and its binding to a dioxin response element.

P A Harper, J V Giannone, A B Okey and M S Denison
Molecular Pharmacology October 1, 1992, 42 (4) 603-612;

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Abstract

In vitro transformation of the human Ah receptor and its binding to a dioxin response element.

P A Harper, J V Giannone, A B Okey and M S Denison
Molecular Pharmacology October 1, 1992, 42 (4) 603-612;
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