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Departments of Endocrinology and Reproduction (C.A.B., A.O.B.) and Pathology (P.D., K.S., J.T.) Erasmus University 3000 DR Rotterdam, The Netherlands
Previous studies in yeast and mammalian cells showed a functional interaction between the amino-terminal domain and the carboxy-terminal, ligand-binding domain (LBD) of the human androgen receptor (AR). In the present study, the AR subdomains involved in this in vivo interaction were determined in more detail. Cotransfection experiments in Chinese hamster ovary (CHO) cells and two-hybrid experiments in yeast revealed that two regions in the NH2-terminal domain are involved in the functional interaction with the LBD: an interacting domain at the very NH2 terminus, located between amino acid residues 3 and 36, and a second domain, essential for transactivation, located between residues 370 and 494. Substitution of glutamic acid by glutamine at position 888 (E888Q) in the AF-2 activation domain (AD) core region in the LBD, markedly decreased the interaction with the NH2-terminal domain. This mutation neither influenced hormone binding nor LBD homodimerization, suggesting a role of the AF-2 AD core region in the functional interaction between the NH2-terminal domain and the LBD. The AF-2 AD core region was also involved in the interaction with the coactivator TIF2 (transcriptional intermediary factor 2), as the E888Q mutation decreased the stimulatory effect of TIF2 on AR AF-2 activity. Cotransfection of TIF2 and the AR NH2-terminal domain expression vectors did not result in synergy between both factors in the induction of AR AF-2 activity. TIF2 highly induced AR AF-2 activity on a complex promoter [mouse mammary tumor virus (MMTV)], but it was hardly active on a minimal promoter (GRE-TATA). In contrast, the AR NH2-terminal domain induced AR AF-2 activity on both promoter constructs. These data indicate that both the AR NH2-terminal domain and the coactivator TIF2 functionally interact, either directly or indirectly, with the AF-2 AD core region in the AR-LBD, but the level of transcriptional response induced by TIF2 depends on the promoter context.
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C.-y. Chang, J. D. Norris, H. Gron, L. A. Paige, P. T. Hamilton, D. J. Kenan, D. Fowlkes, and D. P. McDonnell Dissection of the LXXLL Nuclear Receptor-Coactivator Interaction Motif Using Combinatorial Peptide Libraries: Discovery of Peptide Antagonists of Estrogen Receptors alpha and beta Mol. Cell. Biol., December 1, 1999; 19(12): 8226 - 8239. [Abstract] [Full Text] [PDF]
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C. L. Bevan, S. Hoare, F. Claessens, D. M. Heery, and M. G. Parker The AF1 and AF2 Domains of the Androgen Receptor Interact with Distinct Regions of SRC1 Mol. Cell. Biol., December 1, 1999; 19(12): 8383 - 8392. [Abstract] [Full Text] [PDF]
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D. Szapary, Y. Huang, and S. S. Simons Jr. Opposing Effects of Corepressor and Coactivators in Determining the Dose-Response Curve of Agonists, and Residual Agonist Activity of Antagonists, for Glucocorticoid Receptor-Regulated Gene Expression Mol. Endocrinol., December 1, 1999; 13(12): 2108 - 2121. [Abstract] [Full Text]
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P. Alen, F. Claessens, G. Verhoeven, W. Rombauts, and B. Peeters The Androgen Receptor Amino-Terminal Domain Plays a Key Role in p160 Coactivator-Stimulated Gene Transcription Mol. Cell. Biol., September 1, 1999; 19(9): 6085 - 6097. [Abstract] [Full Text] [PDF]
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M. J. Tetel, P. H. Giangrande, S. A. Leonhardt, D. P. McDonnell, and D. P. Edwards Hormone-Dependent Interaction between the Amino- and Carboxyl-Terminal Domains of Progesterone Receptor in Vitro and in Vivo Mol. Endocrinol., June 1, 1999; 13(6): 910 - 924. [Abstract] [Full Text]
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F. Schaufele Regulation of Estrogen Receptor Activation of the Prolactin Enhancer/Promoter by Antagonistic Activation Function-2-Interacting Proteins Mol. Endocrinol., June 1, 1999; 13(6): 935 - 945. [Abstract] [Full Text]
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X. Sui, K. S. Bramlett, M. C. Jorge, D. A. Swanson, A. C. von Eschenbach, and G. Jenster Specific Androgen Receptor Activation by an Artificial Coactivator J. Biol. Chem., April 2, 1999; 274(14): 9449 - 9454. [Abstract] [Full Text] [PDF]
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B. He, J. A. Kemppainen, and E. M. Wilson FXXLF and WXXLF Sequences Mediate the NH2-terminal Interaction with the Ligand Binding Domain of the Androgen Receptor J. Biol. Chem., July 21, 2000; 275(30): 22986 - 22994. [Abstract] [Full Text] [PDF]
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M. I. Gonzalez and D. M. Robins Oct-1 Preferentially Interacts with Androgen Receptor in a DNA-dependent Manner That Facilitates Recruitment of SRC-1 J. Biol. Chem., February 23, 2001; 276(9): 6420 - 6428. [Abstract] [Full Text] [PDF]
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Q. Wang, J. Lu, and E. L. Yong Ligand- and Coactivator-mediated Transactivation Function (AF2) of the Androgen Receptor Ligand-binding Domain Is Inhibited by the Cognate Hinge Region J. Biol. Chem., March 2, 2001; 276(10): 7493 - 7499. [Abstract] [Full Text] [PDF]
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C. E. Petre, Y. B. Wetherill, M. Danielsen, and K. E. Knudsen Cyclin D1: Mechanism and Consequence of Androgen Receptor Co-repressor Activity J. Biol. Chem., January 11, 2002; 277(3): 2207 - 2215. [Abstract] [Full Text] [PDF]
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A. Bubulya, S.-Y. Chen, C. J. Fisher, Z. Zheng, X.-Q. Shen, and L. Shemshedini c-Jun Potentiates the Functional Interaction between the Amino and Carboxyl Termini of the Androgen Receptor J. Biol. Chem., November 21, 2001; 276(48): 44704 - 44711. [Abstract] [Full Text] [PDF]
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