title: Molecular mechanisms of transcriptional control by Rev-erbα: An energetic foundation for reconciling structure and binding with biological function.
abstract: Rev-erbα and β are nuclear receptors that function as transcriptional repressors of genes involved in regulating circadian rhythms, glucose, and cholesterol metabolism and the inflammatory response. Given these key functions, Rev-erbs are important drug targets for treatment of a number of human pathologies, including cancer, heart disease, and type II diabetes. Transcriptional repression by the Rev-erbs involves direct competition with transcriptional activators for target sites, but also recruitment by the Rev-erbs of the NCoR corepressor protein. Interestingly, Rev-erbs do not appear to interact functionally with a very similar corepressor, Smrt. Transcriptional repression by Rev-erbs is thought to occur in response to the binding of heme, although structural, and ligand binding studies in vitro show that heme and corepressor binding are antagonistic. We carried out systematic studies of the ligand and corepressor interactions to address the molecular basis for corepressor specificity and the energetic consequences of ligand binding using a variety of biophysical approaches. Highly quantitative fluorescence anisotropy assays in competition mode revealed that the Rev-erb specificity for the NCoR corepressor lies in the first two residues of the β-strand in Interaction Domain 1 of NCoR. Our studies confirmed and quantitated the strong antagonism of heme and corepressor binding and significant stabilization of the corepressor complex by a synthetic ligand in vitro. We propose a model which reconciles the contradictory observations concerning the effects of heme binding in vitro and in live cells.
authors.affiliation: Centre de Biochimie Structurale CNRS UMR 5048, INSERM UMR 1054, Université de Montpellier, 34090, Montpellier Cedex, France.
authors.forename: Anaïs
authors.initials: A
authors.lastname: Vaissière
authors.affiliation: Biotechnologie, Pharmacologie Moléculaire et Cellulaire, PEX BCB, Institut de Recherches SERVIER, 125 Chemin de Ronde, 78290, Croissy-sur-Seine, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Berger
authors.affiliation: Centre de Biochimie Structurale CNRS UMR 5048, INSERM UMR 1054, Université de Montpellier, 34090, Montpellier Cedex, France.
authors.forename: Deborah
authors.initials: D
authors.lastname: Harrus
authors.affiliation: Pole d'Innovation Thérapeutique, Recherche et Découverte Métabolisme, Institut de Recherches SERVIER, 11, rue des Moulineaux, 92150, Suresnes, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Dacquet
authors.affiliation: Centre de Biochimie Structurale CNRS UMR 5048, INSERM UMR 1054, Université de Montpellier, 34090, Montpellier Cedex, France.
authors.forename: Albane
authors.initials: A
authors.lastname: Le Maire
authors.affiliation: Biotechnologie, Pharmacologie Moléculaire et Cellulaire, PEX BCB, Institut de Recherches SERVIER, 125 Chemin de Ronde, 78290, Croissy-sur-Seine, France.
authors.forename: Jean A
authors.initials: JA
authors.lastname: Boutin
authors.affiliation: Biotechnologie, Pharmacologie Moléculaire et Cellulaire, PEX BCB, Institut de Recherches SERVIER, 125 Chemin de Ronde, 78290, Croissy-sur-Seine, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Ferry
authors.affiliation: Centre de Biochimie Structurale CNRS UMR 5048, INSERM UMR 1054, Université de Montpellier, 34090, Montpellier Cedex, France.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Royer
mesh_terms: D000595:Amino Acid Sequence; D006418:Heme / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D056971:Nuclear Receptor Co-Repressor 1 / Q000737:chemistry / Q000378:metabolism*; D056953:Nuclear Receptor Subfamily 1, Group D, Member 1 / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D060066:Protein Interaction Maps; D013816:Thermodynamics; D015533:Transcriptional Activation
chemical_list: C539127:NCOR1 protein, human; C064353:NR1D1 protein, human; D056971:Nuclear Receptor Co-Repressor 1; D056953:Nuclear Receptor Subfamily 1, Group D, Member 1; D010455:Peptides; D006418:Heme
keywords: Rev-erb; anisotropy; binding; corepressor; heme