Abstract
Repression of gene transcription by nuclear receptors is mediated by interactions with co-repressor proteins such as SMRT and N-CoR1,2, which in turn recruit histone deacetylases to the chromatin3,4,5. Aberrant interactions between nuclear receptors and co-repressors contribute towards acute promyelocytic leukaemia and thyroid hormone resistance syndrome6,7,8. The binding of co-repressors to nuclear receptors occurs in the unliganded state, and can be stabilized by antagonists9. Here we report the crystal structure of a ternary complex containing the peroxisome proliferator-activated receptor-α ligand-binding domain bound to the antagonist GW6471 and a SMRT co-repressor motif. In this structure, the co-repressor motif adopts a three-turn α-helix that prevents the carboxy-terminal activation helix (AF-2) of the receptor from assuming the active conformation. Binding of the co-repressor motif is further reinforced by the antagonist, which blocks the AF-2 helix from adopting the active position. Biochemical analyses and structure-based mutagenesis indicate that this mode of co-repressor binding is highly conserved across nuclear receptors.
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References
Horlein, A. J. et al. Ligand-independent repression by the thyroid hormone receptor mediated by a nuclear receptor co-repressor. Nature 377, 397–404 (1995).
Chen, J. D. & Evans, R. M. A transcriptional co-repressor that interacts with nuclear hormone receptors. Nature 377, 454–457 (1995).
Nagy, L. et al. Nuclear receptor repression mediated by a complex containing SMRT, mSin3A, and histone deacetylase. Cell 89, 373–380 (1997).
Hassig, C. A., Fleischer, T. C., Billin, A. N., Schreiber, S. L. & Ayer, D. E. Histone deacetylase activity is required for full transcriptional repression by mSin3A. Cell 89, 341–347 (1997).
Laherty, C. D. et al. Histone deacetylases associated with the mSin3 corepressor mediate mad transcriptional repression. Cell 89, 349–356 (1997).
Hong, S. H., David, G., Wong, C. W., Dejean, A. & Privalsky, M. L. SMRT corepressor interacts with PLZF and with the PML-retinoic acid receptor alpha (RARα) and PLZF-RARα oncoproteins associated with acute promyelocytic leukemia. Proc. Natl Acad. Sci. USA 94, 9028–9033 (1997).
Grignani, F. et al. Fusion proteins of the retinoic acid receptor-α recruit histone deacetylase in promyelocytic leukaemia. Nature 391, 815–881 (1998).
Yoh, S. M., Chatterjee, V. K. & Privalsky, M. L. Thyroid hormone resistance syndrome manifests as an aberrant interaction between mutant T3 receptors and transcriptional corepressors. Mol. Endocrinol. 11, 470–480 (1997).
Jackson, T. A. et al. The partial agonist activity of antagonist-occupied steroid receptors is controlled by a novel hinge domain-binding coactivator L7/SPA and the corepressors N-CoR or SMRT. Mol. Endocrinol. 11, 693–705 (1997).
Issemann, I. & Green, S. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators. Nature 347, 645–650 (1990).
Xu, H. E. et al. Structural determinants of ligand binding selectivity between the peroxisome proliferator-activated receptors. Proc. Natl Acad. Sci. USA 98, 13919–13924 (2001).
Onate, S. A., Tsai, S. Y., Tsai, M. J. & O'Malley, B. W. Sequence and characterization of a coactivator for the steroid hormone receptor superfamily. Science 270, 1354–1357 (1995).
Shiau, A. K. et al. The structural basis of estrogen receptor/coactivator recognition and the antagonism of this interaction by tamoxifen. Cell 95, 927–937 (1998).
Darimont, B. D. et al. Structure and specificity of nuclear receptor–coactivator interactions. Genes Dev. 12, 3343–3356 (1998).
Gampe, R. T. Jr et al. Asymmetry in the PPARγ/RXRα crystal structure reveals the molecular basis of heterodimerization among nuclear receptors. Mol. Cell 5, 545–555 (2000).
Nolte, R. T. et al. Ligand binding and co-activator assembly of the peroxisome proliferator-activated receptor-γ. Nature 395, 137–143 (1998).
Heery, D. M., Kalkhoven, E., Hoare, S. & Parker, M. G. A signature motif in transcriptional co-activators mediates binding to nuclear receptors. Nature 387, 733–736 (1997).
Yang, W., Rachez, C. & Freedman, L. P. Discrete roles for peroxisome proliferator-activated receptor gamma and retinoid X receptor in recruiting nuclear receptor coactivators. Mol. Cell Biol. 20, 8008–8017 (2000).
Nagy, L. et al. Mechanism of corepressor binding and release from nuclear hormone receptors. Genes Dev. 13, 3209–3216 (1999).
Hu, X. & Lazar, M. A. The CoRNR motif controls the recruitment of corepressors by nuclear hormone receptors. Nature 402, 93–96 (1999).
Perissi, V. et al. Molecular determinants of nuclear receptor–corepressor interaction. Genes Dev. 13, 3198–3208 (1999).
Zhou, G. et al. Nuclear receptors have distinct affinities for coactivators: characterization by fluorescence resonance energy transfer. Mol. Endocrinol. 12, 1594–1604 (1998).
Xu, H. E. et al. Molecular recognition of fatty acids by peroxisome proliferator-activated receptors. Mol. Cell 3, 397–403 (1999).
Otwinowski, Z. & Minor, W. in Macromolecular Crystallography (eds Carter, J. C. W. & Sweet, R. M.) 307–326 (Academic, New York, 1997).
Navaza, J., Gover, S. & Wolf, W. in Molecular Replacement: Proceedings of the CCP4 Study Weekend (ed. Dodson, E. J.) 87–90 (SERC, Daresbury, 1992).
Cowtan, K. in Joint CCP4 and ESF-EACBM Newsletter on Protein Crystallography 31, 34–38 (1994).
Nolte, R. T., Conlin, R. M., Harrison, S. C. & Brown, R. S. Differing roles for zinc fingers in DNA recognition: structure of a six-finger transcription factor IIIA complex. Proc. Natl Acad. Sci. USA 95, 2938–2943 (1998).
Brunger, A. T. et al. Crystallography & NMR system: A new software suite for macromolecular structure determination. Acta Crystallogr. D 54, 905–921 (1998).
Oberfield, J. L. et al. A peroxisome proliferator-activated receptor gamma ligand inhibits adipocyte differentiation. Proc. Natl Acad. Sci. USA 96, 6102–6106 (1999).
Ullman, E. F. et al. Luminescent oxygen channeling immunoassay: measurement of particle binding kinetics by chemiluminescence. Proc. Natl Acad. Sci. USA 91, 5426–5430 (1994).
Acknowledgements
We thank B. Wisely and R. Bledsoe for making co-repressor expression constructs in early crystallization studies; W. Burkart and M. Moyer for protein sequencing; M. Iannone for compound characterizations; G. Waitt and C. Wagner for mass spectroscopy and amino-acid content analysis; and J. Chrzas and A. Howard for assistance with data collections at 17-ID. Use of the Advanced Photon Source was supported by the US Department of Energy, Basic Energy Sciences, and Office of Science. We also thank L. Kuyper and D. Eggleston for support and critical reading of the manuscript.
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Xu, H., Stanley, T., Montana, V. et al. Structural basis for antagonist-mediated recruitment of nuclear co-repressors by PPARα. Nature 415, 813–817 (2002). https://doi.org/10.1038/415813a
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DOI: https://doi.org/10.1038/415813a
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