Abstract
Retinoids regulate gene expression through binding to the nuclear retinoic acid receptors (RARs) and retinoid X receptors (RXRs). In contrast, no ligands for the retinoic acid receptor–related orphan receptors β and γ (RORβ and γ) have been identified, yet structural data and structure-function analyses indicate that RORβ is a ligand-regulated nuclear receptor. Using nondenaturing mass spectrometry and scintillation proximity assays we found that all-trans retinoic acid (ATRA) and several retinoids bind to the RORβ ligand-binding domain (LBD). The crystal structures of the complex with ATRA and with the synthetic analog ALRT 1550 reveal the binding modes of these ligands. ATRA and related retinoids inhibit RORβ but not RORα transcriptional activity suggesting that high-affinity, subtype-specific ligands could be designed for the identification of RORβ target genes. Our results identify RORβ as a retinoid-regulated nuclear receptor, providing a novel pathway for retinoid action.
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Acknowledgements
We thank M. Salvati from Bristol-Myers Squibb for the gift of ALRT 1550 and M. Klaus from Roche for the gift of RO 41-5253 and all-trans-4-oxo retinoic acid, P. Eberling for peptide synthesis, B. Blumberg for advice in performing scintillation proximity assays, P. Carpentier for support on ESRF beamline BM14 CRG and C. Schulze-Briese for support on the SLS X065A beamline, J. Cavarelli for help with the ALRT-complex data, and G. Tocchini-Valentini, V. Lamour and P. Hublitz for useful discussions. This work was supported by grants from the Deutsche Forschungsgemeinschaft to R.S. and by funds from the Centre National de la Recherche Scientifique, the Institut National de la Santé et de la Recherche Médicale, Université Louis Pasteur de Strasbourg and the Fond National de la Science to the Strasbourg Génopole. We acknowledge the financial support of the European Union for the international research project SPINE. S.S. acknowledges a grant from the CNRS and Eli Lilly.
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Stehlin-Gaon, C., Willmann, D., Zeyer, D. et al. All-trans retinoic acid is a ligand for the orphan nuclear receptor RORβ. Nat Struct Mol Biol 10, 820–825 (2003). https://doi.org/10.1038/nsb979
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DOI: https://doi.org/10.1038/nsb979
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