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M410456200v1
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Papers In Press, published online ahead of print October 13, 2004
J. Biol. Chem, 10.1074/jbc.M410456200
Submitted on September 10, 2004
Revised on October 8, 2004
Accepted on October 8, 2004

Recruitment of thyroid hormone receptor/retinoblastoma interacting protein 230 (TRIP230) by the Aryl hydrocarbon receptor nuclear translocator (ARNT) is essential for the transcriptional response to both dioxin and hypoxia

Timothy V. Beischlag, Robert T. Taylor, David W. Rose, Diana Y. Yoon, Yumay Chen, Wen-Hwa Lee, Michael G. Rosenfeld, and Oliver Hankinson

Pathology and Laboratory Medicine, UCLA, Los Angeles, CA 90095-1732

Corresponding Author: ohank{at}mednet.ucla.edu

The aryl hydrocarbon receptor nuclear translocator (ARNT/HIF1b) mediates an organism’s response to various environmental cues, including those to chemical carcinogens, such as 2,3,7,8-tetrachlorodibenzo-r-dioxin (TCDD or dioxin), via its formation of a functional transcription factor with the ligand activated aryl hydrocarbon receptor (AHR). Similarly, tissue responses to hypoxia are largely mediated through the hypoxia inducible factor (HIF-1) heterodimeric transcription factor, comprising hypoxia-inducible factor-1a (HIF-1a) and ARNT. The latter response is essential for a metabolic switch from oxidative phosphorylation to glycolytic anaerobic metabolism, as well as for angiogenesis, and has been implicated as necessary for growth in many solid tumors. In this report, we demonstrate that the thyroid hormone receptor/retinoblastoma interacting protein 230 (TRIP230) interacts directly with ARNT and is essential for both hypoxic and TCDD mediated transcriptional responses. We initially identified TRIP230 as an ARNT-interacting protein in a yeast two-hybrid assay screen. This interaction was confirmed in mammalian cell systems using co-immunoprecipitation, and in mammalian two-hybrid assays. Furthermore, TRIP230 could be recorded at sites of activated transcription of either TCDD or hypoxia inducible genes in a stimulus-dependent fashion by chromatin immunoprecipitation analysis. Finally, using single-cell micro-injection and RNA interference assays we demonstrate that TRIP230 is indispensable for TCDD- and hypoxia-dependent gene transcription.


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