This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Tetel, M. J. Articles by Edwards, D. P. Search for Related Content PubMed PubMed Citation Articles by Tetel, M. J. Articles by Edwards, D. P.

Hinge and Amino-Terminal Sequences Contribute to Solution Dimerization of Human Progesterone Receptor

Department of Pathology and Program in Molecular Biology (M.J.T., T.L., D.P.E.) University of Colorado Health Sciences Center Denver, Colorado 80262
Department of Physiology (S.J., P.C., D.F.S.) Wayne State University School of Medicine, Detroit, Michigan 48201

We and others have shown previously that progesterone receptors (PR) form homodimers in solution in the absence of DNA and that dimers are the preferential form of receptor that binds with high affinity to target DNA. To determine the sequence regions involved in solution homodimerization, wild type PR and truncated PR proteins were expressed in an insect baculovirus system. The expression constructs included the ligand-binding domain [LBD, amino acids (aa) 688–933], the LBD plus hinge (hLBD, aa 634–933), the hLBD plus the DNA-binding domain (DhLBD, aa 538–933), and the full- length A and B isoforms of PR. PR-PR interactions were detected by three methods, coimmunoprecipitation of the PR fragments with full-length PR-A, pull-down of PR-polypeptides with polyhistidine-tagged versions of the same polypeptides immobilized to metal affinity columns and cooperative ligand-binding assays (Hill coefficient, n_H > 1 indicating PR-PR interaction). By all three assays, the LBD alone was not sufficient to mediate protein-protein interaction. However, the LBD did exhibit other properties ascribed to this domain, including binding to steroids with a relatively good affinity and specificity, ligand-induced conformational changes at the carboxyl terminus tail and binding of heat shock protein 90 and its dissociation in response to hormone. Thus, failure of the expressed LBD to mediate dimerization does not appear to be due to an extensively misfolded or unstable polypeptide. The minimal carboxyl-terminal fragment capable of mediating PR-PR interaction was the hLBD construct. However, by immobilized metal affinity chromatography assay, self-association of PR-A was 3.5-fold more efficient than that of either the DhLBD or hLBD constructs. An expressed amino-terminal domain (aa 165–535) lacking the DNA-binding domain, hinge, and LBD was found to physically associate with PR-A or with another amino-terminal fragment lacking the LBD, but retaining the DNA-binding domain. These results provide evidence for direct amino-terminal interactions in the more efficient PR-PR interaction exhibited by wild-type PR-A, as compared with DhLBD and hLBD constructs. The overall results of this paper are consistent with the conclusion that the carboxyl-terminal LBD is not sufficient for mediating PR dimerization and that multiple regions, including the hinge and amino-terminal sequences, contribute either directly or indirectly to homodimerization of PR.

This article has been cited by other articles:

				J. D. Graham, A. R. Hanson, A. J. Croft, A. H. Fox, and C. L. Clarke Nuclear matrix binding is critical for progesterone receptor movement into nuclear foci FASEB J, February 1, 2009; 23(2): 546 - 556. [Abstract] [Full Text] [PDF]

				A. Romano, B. Delvoux, D.-C. Fischer, and P. Groothuis The PROGINS polymorphism of the human progesterone receptor diminishes the response to progesterone J. Mol. Endocrinol., February 1, 2007; 38(2): 331 - 350. [Abstract] [Full Text] [PDF]

				Y. Wu, H. Kawate, K. Ohnaka, H. Nawata, and R. Takayanagi Nuclear Compartmentalization of N-CoR and Its Interactions with Steroid Receptors. Mol. Cell. Biol., September 1, 2006; 26(17): 6633 - 6655. [Abstract] [Full Text] [PDF]

				Y. Ma, P. Katiyar, L. P. Jones, S. Fan, Y. Zhang, P. A. Furth, and E. M. Rosen The Breast Cancer Susceptibility Gene BRCA1 Regulates Progesterone Receptor Signaling in Mammary Epithelial Cells Mol. Endocrinol., January 1, 2006; 20(1): 14 - 34. [Abstract] [Full Text] [PDF]

				T. M. Price, E. L. Hansen, and T. N. Oliver Immunofluorescent Localization of a Novel Progesterone Receptor(s) in a T47D-Y Breast Cancer Cell Line Lacking Genomic Progesterone Receptor Expression Reproductive Sciences, December 1, 2005; 12(8): 610 - 616. [Abstract] [PDF]

				S. Cho, B. L. Kagan, J. A. Blackford Jr., D. Szapary, and S. S. Simons Jr. Glucocorticoid Receptor Ligand Binding Domain Is Sufficient for the Modulation of Glucocorticoid Induction Properties by Homologous Receptors, Coactivator Transcription Intermediary Factor 2, and Ubc9 Mol. Endocrinol., February 1, 2005; 19(2): 290 - 311. [Abstract] [Full Text] [PDF]

				I. U. Agoulnik, X.-W. Tong, D.-C. Fischer, K. Korner, N. E. Atkinson, D. P. Edwards, D. R. Headon, N. L. Weigel, and D. G. Kieback A Germline Variation in the Progesterone Receptor Gene Increases Transcriptional Activity and May Modify Ovarian Cancer Risk J. Clin. Endocrinol. Metab., December 1, 2004; 89(12): 6340 - 6347. [Abstract] [Full Text] [PDF]

				I. U. Agoulnik, W. C. Krause, W. E. Bingman III, H. T. Rahman, M. Amrikachi, G. E. Ayala, and N. L. Weigel Repressors of Androgen and Progesterone Receptor Action J. Biol. Chem., August 15, 2003; 278(33): 31136 - 31148. [Abstract] [Full Text] [PDF]

				B. Larsson and I. Nemere Effect of Growth and Maturation on Membrane-Initiated Actions of 1,25-Dihydroxyvitamin D3. I. Calcium Transport, Receptor Kinetics, and Signal Transduction in Intestine of Male Chickens Endocrinology, May 1, 2003; 144(5): 1726 - 1735. [Abstract] [Full Text] [PDF]

				P. S. Quadros, J. L. Pfau, A. Y. N. Goldstein, G. J. De Vries, and C. K. Wagner Sex Differences in Progesterone Receptor Expression: A Potential Mechanism for Estradiol-Mediated Sexual Differentiation Endocrinology, October 1, 2002; 143(10): 3727 - 3739. [Abstract] [Full Text] [PDF]

				S. E. Wardell, V. Boonyaratanakornkit, J. S. Adelman, A. Aronheim, and D. P. Edwards Jun Dimerization Protein 2 Functions as a Progesterone Receptor N-Terminal Domain Coactivator Mol. Cell. Biol., August 1, 2002; 22(15): 5451 - 5466. [Abstract] [Full Text] [PDF]

				B. He, N. T. Bowen, J. T. Minges, and E. M. Wilson Androgen-induced NH2- and COOH-terminal Interaction Inhibits p160 Coactivator Recruitment by Activation Function 2 J. Biol. Chem., November 2, 2001; 276(45): 42293 - 42301. [Abstract] [Full Text] [PDF]

				J. G. A. Savory, G. G. Préfontaine, C. Lamprecht, M. Liao, R. F. Walther, Y. A. Lefebvre, and R. J. G. Haché Glucocorticoid Receptor Homodimers and Glucocorticoid-Mineralocorticoid Receptor Heterodimers Form in the Cytoplasm through Alternative Dimerization Interfaces Mol. Cell. Biol., February 1, 2001; 21(3): 781 - 793. [Abstract] [Full Text]

				J.-a. Tan, S. H. Hall, P. Petrusz, and F. S. French Thyroid Receptor Activator Molecule, TRAM-1, Is an Androgen Receptor Coactivator Endocrinology, September 1, 2000; 141(9): 3440 - 3450. [Abstract] [Full Text] [PDF]

				D. L. Bain, M. A. Franden, J. L. McManaman, G. S. Takimoto, and K. B. Horwitz The N-terminal Region of the Human Progesterone A-receptor. STRUCTURAL ANALYSIS AND THE INFLUENCE OF THE DNA BINDING DOMAIN J. Biol. Chem., March 15, 2000; 275(10): 7313 - 7320. [Abstract] [Full Text] [PDF]

				D. L. Clemm, L. Sherman, V. Boonyaratanakornkit, W. T. Schrader, N. L. Weigel, and D. P. Edwards Differential Hormone-Dependent Phosphorylation of Progesterone Receptor A and B Forms Revealed by a Phosphoserine Site-Specific Monoclonal Antibody Mol. Endocrinol., January 1, 2000; 14(1): 52 - 65. [Abstract] [Full Text]

				G. Giannoukos, A. M. Silverstein, W. B. Pratt, and S. S. Simons Jr. The Seven Amino Acids (547-553) of Rat Glucocorticoid Receptor Required for Steroid and Hsp90 Binding Contain a Functionally Independent LXXLL Motif That Is Critical for Steroid Binding J. Biol. Chem., December 17, 1999; 274(51): 36527 - 36536. [Abstract] [Full Text] [PDF]

				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]

				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]

				C. S. Lim, C. T. Baumann, H. Htun, W. Xian, M. Irie, C. L. Smith, and G. L. Hager Differential Localization and Activity of the A- and B-Forms of the Human Progesterone Receptor Using Green Fluorescent Protein Chimeras Mol. Endocrinol., March 1, 1999; 13(3): 366 - 375. [Abstract] [Full Text]

				J. A. Kemppainen, E. Langley, C.-i. Wong, K. Bobseine, W. R. Kelce, and E. M. Wilson Distinguishing Androgen Receptor Agonists and Antagonists: Distinct Mechanisms of Activation by Medroxyprogesterone Acetate and Dihydrotestosterone Mol. Endocrinol., March 1, 1999; 13(3): 440 - 454. [Abstract] [Full Text]

				S. A. Leonhardt, M. Altmann, and D. P. Edwards Agonist and Antagonists Induce Homodimerization and Mixed Ligand Heterodimerization of Human Progesterone Receptors in Vivo by a Mammalian Two-Hybrid Assay Mol. Endocrinol., December 1, 1998; 12(12): 1914 - 1930. [Abstract] [Full Text]

				P. H. Giangrande, G. Pollio, and D. P. McDonnell Mapping and Characterization of the Functional Domains Responsible for the Differential Activity of the A and B Isoforms of the Human Progesterone Receptor J. Biol. Chem., December 26, 1997; 272(52): 32889 - 32900. [Abstract] [Full Text] [PDF]

				D. F. Skafar, R. Xu, J. Morales, J. Ram, and J. R. Sowers Female Sex Hormones and Cardiovascular Disease in Women J. Clin. Endocrinol. Metab., December 1, 1997; 82(12): 3913 - 3918. [Abstract] [Full Text] [PDF]

				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]

				R. C. J. Ribeiro, W. Feng, R. L. Wagner, C. H. R. M. Costa, A. C. Pereira, J. W. Apriletti, R. J. Fletterick, and J. D. Baxter Definition of the Surface in the Thyroid Hormone Receptor Ligand Binding Domain for Association as Homodimers and Heterodimers with Retinoid X Receptor J. Biol. Chem., April 27, 2001; 276(18): 14987 - 14995. [Abstract] [Full Text] [PDF]

				K. Takahashi, T. Taira, T. Niki, C. Seino, S. M. M. Iguchi-Ariga, and H. Ariga DJ-1 Positively Regulates the Androgen Receptor by Impairing the Binding of PIASxalpha to the Receptor J. Biol. Chem., September 28, 2001; 276(40): 37556 - 37563. [Abstract] [Full Text] [PDF]

				D. L. Bain, M. A. Franden, J. L. McManaman, G. S. Takimoto, and K. B. Horwitz The N-terminal Region of Human Progesterone B-receptors. BIOPHYSICAL AND BIOCHEMICAL COMPARISON TO A-RECEPTORS J. Biol. Chem., June 22, 2001; 276(26): 23825 - 23831. [Abstract] [Full Text] [PDF]