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Title: Ligand-dependent interactions of the Ah receptor with coactivators in a mammalian two-hybrid assay

Abstract

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a high affinity ligand for the aryl hydrocarbon receptor (AhR). In this study, we investigated structure-dependent differences in activation of the AhR by a series of halogenated aromatic hydrocarbons. TCDD, 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PeCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 2,3,4,7,8-pentachlorodibenzofuran (PeCDF), and 3,3',4,4',5-pentachlorobiphenyl (PCB126) induced CYP1A1-dependent activities in HEK293 human embryonic kidney, Panc1 pancreatic cancer, and Hepa1c1c7 mouse hepatoma cell lines. There was a structure-dependent difference in the efficacy of TCDF and PCB126 in HEK293 and Panc1 cells since induced CYP1A1 mRNA levels were lower than observed for the other congeners. A mammalian two-hybrid assay in cells transfected with GAL4-coactivator and AhR-VP16 chimeras was used to investigate structure-dependent interactions of these chimeras in Panc1, HEK293, and Hepa1c1c7 cells. The reporter construct pGAL4-luc contains five tandem GAL4 response elements linked to the luciferase gene and the GAL4-coactivator chimeras express several coactivators including steroid receptor coactivator 1 (SRC-1), SRC-2 and SRC-3, the mediator coactivator TRAP220, coactivator associated arginine methyl transferase 1 (CARM-1), and peroxisome proliferator-activated receptor {gamma} coactivator 1 (PGC-1). Results of the mammalian two-hybrid studies clearly demonstrate that activation of pGAL4-luc in cells transfected with VP-AhR and GAL4-coactivator chimeras is dependent on the structure of the HAH congener, cell context, and coactivator, suggestingmore » that the prototypical HAH congeners used in this study exhibit selective AhR modulator activity.« less

Authors:
 [1];  [2];  [1]
  1. Department of Veterinary Physiology and Pharmacology, Texas A and M University, College Station, TX 77843 (United States)
  2. Dow Chemical Company, Toxicology and Environmental Research and Consulting, Midland, MI 48674 (United States)
Publication Date:
OSTI Identifier:
21077934
Resource Type:
Journal Article
Journal Name:
Toxicology and Applied Pharmacology
Additional Journal Information:
Journal Volume: 227; Journal Issue: 2; Other Information: DOI: 10.1016/j.taap.2007.10.019; PII: S0041-008X(07)00482-6; Copyright (c) 2007 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0041-008X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ARGININE; CHIMERAS; DIOXIN; HALOGENATED AROMATIC HYDROCARBONS; HEPATOMAS; HYBRIDIZATION; HYDROCARBONS; KIDNEYS; LIGANDS; LUCIFERASE; MICE; PANCREAS; RECEPTORS; STEROIDS

Citation Formats

Shu, Zhang, Rowlands, Craig, Safe, Stephen, and Institute of Biosciences and Technology, Texas A and M University Health Science Center, Houston, TX 77030. Ligand-dependent interactions of the Ah receptor with coactivators in a mammalian two-hybrid assay. United States: N. p., 2008. Web. doi:10.1016/j.taap.2007.10.019.
Shu, Zhang, Rowlands, Craig, Safe, Stephen, & Institute of Biosciences and Technology, Texas A and M University Health Science Center, Houston, TX 77030. Ligand-dependent interactions of the Ah receptor with coactivators in a mammalian two-hybrid assay. United States. https://doi.org/10.1016/j.taap.2007.10.019
Shu, Zhang, Rowlands, Craig, Safe, Stephen, and Institute of Biosciences and Technology, Texas A and M University Health Science Center, Houston, TX 77030. 2008. "Ligand-dependent interactions of the Ah receptor with coactivators in a mammalian two-hybrid assay". United States. https://doi.org/10.1016/j.taap.2007.10.019.
@article{osti_21077934,
title = {Ligand-dependent interactions of the Ah receptor with coactivators in a mammalian two-hybrid assay},
author = {Shu, Zhang and Rowlands, Craig and Safe, Stephen and Institute of Biosciences and Technology, Texas A and M University Health Science Center, Houston, TX 77030},
abstractNote = {2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a high affinity ligand for the aryl hydrocarbon receptor (AhR). In this study, we investigated structure-dependent differences in activation of the AhR by a series of halogenated aromatic hydrocarbons. TCDD, 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PeCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 2,3,4,7,8-pentachlorodibenzofuran (PeCDF), and 3,3',4,4',5-pentachlorobiphenyl (PCB126) induced CYP1A1-dependent activities in HEK293 human embryonic kidney, Panc1 pancreatic cancer, and Hepa1c1c7 mouse hepatoma cell lines. There was a structure-dependent difference in the efficacy of TCDF and PCB126 in HEK293 and Panc1 cells since induced CYP1A1 mRNA levels were lower than observed for the other congeners. A mammalian two-hybrid assay in cells transfected with GAL4-coactivator and AhR-VP16 chimeras was used to investigate structure-dependent interactions of these chimeras in Panc1, HEK293, and Hepa1c1c7 cells. The reporter construct pGAL4-luc contains five tandem GAL4 response elements linked to the luciferase gene and the GAL4-coactivator chimeras express several coactivators including steroid receptor coactivator 1 (SRC-1), SRC-2 and SRC-3, the mediator coactivator TRAP220, coactivator associated arginine methyl transferase 1 (CARM-1), and peroxisome proliferator-activated receptor {gamma} coactivator 1 (PGC-1). Results of the mammalian two-hybrid studies clearly demonstrate that activation of pGAL4-luc in cells transfected with VP-AhR and GAL4-coactivator chimeras is dependent on the structure of the HAH congener, cell context, and coactivator, suggesting that the prototypical HAH congeners used in this study exhibit selective AhR modulator activity.},
doi = {10.1016/j.taap.2007.10.019},
url = {https://www.osti.gov/biblio/21077934}, journal = {Toxicology and Applied Pharmacology},
issn = {0041-008X},
number = 2,
volume = 227,
place = {United States},
year = {Sat Mar 01 00:00:00 EST 2008},
month = {Sat Mar 01 00:00:00 EST 2008}
}