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PMID: 2833743 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Inducible, receptor-dependent protein-DNA interactions at a dioxin-responsive transcriptional enhancer.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 8 ·1988-04-00 ·Pages 2528-32

Denison MS, Fisher JM, Whitlock JP

Abstract

We have identified in mouse hepatoma cells a third cis-acting dioxin-responsive element (DRE) within the 5' flanking region of the cytochrome P1-450 gene, which is transcriptionally activated by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The DRE can activate a heterologous promoter and functions in either orientation; therefore, it has the properties of a transcriptional enhancer. The DRE fails to activate transcription in receptor-defective cells; therefore, it requires TCDD-receptor complexes for its function. By using a gel retardation assay, we show that nuclear extracts contain a protein that binds to the DRE in TCDD-inducible, receptor-dependent, and DNA sequence-specific fashion. The protein-DNA interaction occurs within 10 min of exposure of the cell to TCDD and does not require ongoing protein synthesis. Our results imply that the TCDD-receptor complex interacts specifically with the DRE and demonstrate a relationship between protein-DNA interaction in vitro and function in vivo. Our findings also suggest that the affinity of the TCDD-receptor complex for the DRE may be relatively high in comparison to analogous protein-DNA interactions at other inducible enhancers.

MeSH Terms
Animals Cloning, Molecular Cytochrome P-450 Enzyme System/genetics DNA-Binding Proteins/physiology Dioxins/metabolism Enhancer Elements, Genetic In Vitro Techniques Mice Nuclear Proteins/physiology Polychlorinated Dibenzodioxins/metabolism Receptors, Aryl Hydrocarbon Receptors, Drug/metabolism Regulatory Sequences, Nucleic Acid Transcription Factors/physiology
Chemicals
DNA-Binding Proteins Dioxins Nuclear Proteins Polychlorinated Dibenzodioxins Receptors, Aryl Hydrocarbon Receptors, Drug Transcription Factors Cytochrome P-450 Enzyme System
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Denison M S
Department of Pharmacology, Stanford University School of Medicine, CA 94305.
Fisher J M
Whitlock J P
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45 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.8.2528
Published
1988-04-00
Pages
2528-32
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC280030
Subset
IM
Grants
NIEHS NIH HHS · ES 03719 · United States
External Links
PubMed source DOI: 10.1073/pnas.85.8.2528 PMC Full Text View journal details
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