PMID- 18048071 OWN - NLM STAT- MEDLINE DCOM- 20080331 LR - 20211020 IS - 0041-008X (Print) IS - 1096-0333 (Electronic) IS - 0041-008X (Linking) VI - 227 IP - 2 DP - 2008 Mar 1 TI - Ligand-dependent interactions of the Ah receptor with coactivators in a mammalian two-hybrid assay. PG - 196-206 AB - 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. FAU - Zhang, Shu AU - Zhang S AD - Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station, TX 77843, USA. FAU - Rowlands, Craig AU - Rowlands C FAU - Safe, Stephen AU - Safe S LA - eng GR - P42 ES004917/ES/NIEHS NIH HHS/United States GR - P30 ES009106-09/ES/NIEHS NIH HHS/United States GR - P42 ES004917-190007/ES/NIEHS NIH HHS/United States GR - ES09106/ES/NIEHS NIH HHS/United States GR - ES04917/ES/NIEHS NIH HHS/United States GR - P30 ES009106/ES/NIEHS NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20071101 PL - United States TA - Toxicol Appl Pharmacol JT - Toxicology and applied pharmacology JID - 0416575 RN - 0 (Herpes Simplex Virus Protein Vmw65) RN - 0 (Hydrocarbons, Aromatic) RN - 0 (Ligands) RN - 0 (RNA, Messenger) RN - 0 (Receptors, Aryl Hydrocarbon) RN - EC 1.14.14.1 (Cytochrome P-450 CYP1A1) SB - IM MH - Animals MH - Blotting, Western MH - Cell Line MH - Chimera/genetics MH - Cytochrome P-450 CYP1A1/biosynthesis MH - Enzyme Induction/drug effects MH - Herpes Simplex Virus Protein Vmw65/genetics MH - Humans MH - Hydrocarbons, Aromatic/pharmacology MH - Ligands MH - Mice MH - RNA, Messenger/biosynthesis/genetics MH - Receptors, Aryl Hydrocarbon/*drug effects/genetics MH - Reverse Transcriptase Polymerase Chain Reaction MH - Structure-Activity Relationship MH - Transfection PMC - PMC2288743 MID - NIHMS41795 EDAT- 2007/12/01 09:00 MHDA- 2008/04/01 09:00 PMCR- 2009/03/01 CRDT- 2007/12/01 09:00 PHST- 2007/09/10 00:00 [received] PHST- 2007/10/18 00:00 [revised] PHST- 2007/10/19 00:00 [accepted] PHST- 2007/12/01 09:00 [pubmed] PHST- 2008/04/01 09:00 [medline] PHST- 2007/12/01 09:00 [entrez] PHST- 2009/03/01 00:00 [pmc-release] AID - S0041-008X(07)00482-6 [pii] AID - 10.1016/j.taap.2007.10.019 [doi] PST - ppublish SO - Toxicol Appl Pharmacol. 2008 Mar 1;227(2):196-206. doi: 10.1016/j.taap.2007.10.019. Epub 2007 Nov 1.