1. The orientation of the C-terminal domain of the Saccharomyces cerevisiae Rap1 protein is determined by its binding to DNA
- Author
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Rachel Lescasse, Patrick Weber, Javier Pérez, Marie-Hélène Le Du, Simona Miron, Bertrand Raynal, Sylvaine Gasparini, Gabriel David, B. Matot, Sophie Zinn-Justin, Bertrand Castaing, Yann-Vaï Le Bihan, Protéines membranaires transductrices d'énergie (PMTE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Service d'Instabilité Génétique Réparation Recombinaison (SIGRR), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Synchrotron SOLEIL (SSOLEIL), Centre National de la Recherche Scientifique (CNRS), Centre de biophysique moléculaire (CBM), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Cristallogenèse et Diffraction des Rayons X (Plate-forme/PF6), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Biophysique des macromolécules et de leurs interactions (Plate-forme), Institut Pasteur [Paris] (IP), Commissariat à l'Energie Atomique, Centre National de la Recherche Scientifique, French ‘Agence National pour la Recherche’ (ANR-06-BLAN-0076). Funding for open access charge: Commissariat à l'Energie Atomique., ANR-06-BLAN-0076,NHEJ&TELO,Double-strand break repair and genome stability: the Non-Homologous End Joining pathway and its suppression at telomeres(2006), Université d'Orléans (UO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Laboratoire de Physique des Solides (LPS), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Chemistry, Food Chemsitry, University of Hamburg, and Laboratoire de Biologie Structurale et Radiobiologie (LBSR)
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Models, Molecular ,BUDDING YEAST ,Crystallography, X-Ray ,Shelterin Complex ,MESH: Telomere-Binding Proteins ,MESH: Protein Structure, Tertiary ,chemistry.chemical_compound ,MESH: Saccharomyces cerevisiae Proteins ,0302 clinical medicine ,X-Ray Diffraction ,Transcription (biology) ,Structural Biology ,ANGLE SCATTERING DATA ,MESH: Nuclear Magnetic Resonance, Biomolecular ,Silent Information Regulator Proteins, Saccharomyces cerevisiae ,Genetics ,Telomere-binding protein ,0303 health sciences ,[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,biology ,YEAST RAP1 ,MESH: DNA ,MESH: X-Ray Diffraction ,ASSOCIATION ,MESH: Transcription Factors ,SEPARATION ,REPRESSOR ACTIVATOR PROTEIN-1 ,MESH: Models, Molecular ,Binding domain ,endocrine system ,Saccharomyces cerevisiae Proteins ,HMG-box ,Saccharomyces cerevisiae ,Telomere-Binding Proteins ,03 medical and health sciences ,Scattering, Small Angle ,TARGET SITES ,MESH: Silent Information Regulator Proteins, Saccharomyces cerevisiae ,Nuclear Magnetic Resonance, Biomolecular ,MESH: Scattering, Small Angle ,030304 developmental biology ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,DNA-binding domain ,DNA ,MESH: Crystallography, X-Ray ,biology.organism_classification ,Protein Structure, Tertiary ,DNA binding site ,BIOLOGICAL MACROMOLECULES ,enzymes and coenzymes (carbohydrates) ,chemistry ,TELOMERE LENGTH ,Biophysics ,TRANSCRIPTIONAL ACTIVATION ,030217 neurology & neurosurgery ,Transcription Factors - Abstract
International audience; Rap1 is an essential DNA-binding factor from the yeast Saccharomyces cerevisiae involved in transcription and telomere maintenance. Its binding to DNA targets Rap1 at particular loci, and may optimize its ability to form functional macromolecular assemblies. It is a modular protein, rich in large potentially unfolded regions, and comprising BRCT, Myb and RCT well-structured domains. Here, we present the architectures of Rap1 and a Rap1/DNA complex, built through a step-by-step integration of small angle X-ray scattering, X-ray crystallography and nuclear magnetic resonance data. Our results reveal Rap1 structural adjustment upon DNA binding that involves a specific orientation of the C-terminal (RCT) domain with regard to the DNA binding domain (DBD). Crystal structure of DBD in complex with a long DNA identifies an essential wrapping loop, which constrains the orientation of the RCT and affects Rap1 affinity to DNA. Based on our structural information, we propose a model for Rap1 assembly at telomere.
- Published
- 2011
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