Contributors: Institut de biologie structurale (IBS - UMR 5075); Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG); Direction de Recherche Fondamentale (CEA) (DRF (CEA)); Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)); Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA); Integrated Structural Biology Grenoble (ISBG); Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-European Molecular Biology Laboratory Grenoble (EMBL)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA); Institut de Génomique Fonctionnelle (IGF); Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM); Graduate School (EUR) CBH—Chemistry, Biology, and Health; Grenoble Instruct-ERIC center (ISBG); European Synchrotron Radiation Facility; Synchrotron SOLEIL; ANR-12-BSV8-0025,NNViPol,Structure et fonction de la machine de transcription et réplication des virus à ARN non-segmenté de polarité négative(2012); ANR-17-EURE-0003,CBH-EUR-GS,CBH-EUR-GS(2017); ANR-10-INBS-0005,FRISBI,Infrastructure Française pour la Biologie Structurale Intégrée(2010); European Project: 263186,EC:FP7:SPA,FP7-SPACE-2010-1,GRAAL(2011); European Project: DEQ20170336754
نبذة مختصرة : International audience ; As for all non-segmented negative RNA viruses, rabies virus has its genome packaged in a linear assembly of nucleoprotein (N), named nucleocapsid. The formation of new nucleocapsids during virus replication in cells requires the production of soluble N protein in complex with its phosphoprotein (P) chaperone. In this study, we reconstituted a soluble heterodimeric complex between an armless N protein of rabies virus (RABV), lacking its N-terminal subdomain (NNT-ARM), and a peptide encompassing the N$^0$ chaperon module of the P protein. We showed that the chaperone module undergoes a disordered−order transition when it assembles with N$^0$ and measured an affinity in the low nanomolar range using a competition assay. We solved the crystal structure of the complex at a resolution of 2.3 Å, unveiling the details of the conserved interfaces. MD simulations showed that both the chaperon module of P and RNA-mediated polymerization reduced the ability of the RNA binding cavity to open and close. Finally, by reconstituting a complex with full-length P protein, we demonstrated that each P dimer could independently chaperon two N$^0$ molecules.
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