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Department of Medicine (X.X., P.M.Y.), Johns Hopkins Bayview Medical Center, Johns Hopkins University, Baltimore, Maryland 21224; Molecular Regulation and Neuroendocrinology Section (Y.L., S.A., R.Y.), Clinical Endocrinology Branch, National Institute of Diabetes and Digestive and Kidney Diseases and National Heart, Lung, and Blood Institute (M.K.), National Institutes of Health, Bethesda, Maryland 20892; and Department of Physiology and Biophysics (J.F.), University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854
Address all correspondence and requests for reprints to: Paul M. Yen, M.D., Endocrinology Division, Department of Medicine, Johns Hopkins Bayview Medical Center, 4940 Eastern Avenue, Room B114, Baltimore, Maryland 21224. E-mail: pyen3{at}jhmi.edu.
Currently, little is known about the direct interactions of general transcription factors and nuclear hormone receptors. To investigate the potential role of the general transcription factor, TFIIH, in T3-mediated transcriptional activation, we examined thyroid hormone receptor (TR) interaction with individual TFIIH subunits in a yeast-two hybrid system. Among the nine subunits of TFIIH studied, only p62 subunit interacted with TRß in a ligand-dependent manner. Glutathione-S-transferase pull-down and in vivo coimmunoprecipitation studies also demonstrated direct TR/p62 interaction. Using chromatin immunoprecipitation assays, we showed that TFIIH subunits were corecruited on or near an endogenous thyroid hormone response element upon T3 addition. Cotransfection studies with TSA201 cells showed that p62 increased T3-mediated transcription, which could be further enhanced when p62 and another TFIIH subunit, p44, were cotransfected simultaneously. Taken together, these data suggest that TRs can interact directly with a subunit of TFIIH and may provide an alternative pathway for nuclear receptor communication with the general transcription machinery that circumvents coactivators.
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