Picornaviral polymerase domain exchanges reveal a modular basis for distinct biochemical activities of viral RNA-dependent RNA polymerases
| Autor: | Stéphanie Beaucourt, Colleen L. Watkins, Brian J. Kempf, David J. Barton, Olve B. Peersen |
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| Přispěvatelé: | Colorado State University [Fort Collins] (CSU), Populations virales et Pathogenèse - Viral Populations and Pathogenesis, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), This work was supported by National Institutes of Health Grants AI059130 (to O. B. P.) and AI042189 (to D. J. B.)., Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Conformational change viruses Biochemistry RNA-Protein Interaction enzyme kinetics Polymerase viral polymerase biology poliovirus [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] Enterovirus B Human MESH: RNA Viral MESH: RNA-Dependent RNA Polymerase Protein Structure and Folding [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology RNA Viral MESH: Protein Domains MESH: Poliovirus Computational biology virus Coxsackievirus Virus RNA-dependent RNA polymerase (RdRP) Viral Proteins 03 medical and health sciences conformational change Protein Domains [CHIM.CRIS]Chemical Sciences/Cristallography Humans Molecular Biology coxsackievirus MESH: Humans 030102 biochemistry & molecular biology RNA protein engineering Cell Biology Protein engineering RNA-Dependent RNA Polymerase biology.organism_classification MESH: Viral Proteins body regions 030104 developmental biology Viral replication MESH: Enterovirus B Human MESH: HeLa Cells biology.protein viral replication RNA–protein interaction HeLa Cells |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2020, 295 (31), pp.10624-10637. ⟨10.1074/jbc.RA120.013906⟩ Journal of Biological Chemistry, 2020, 295 (31), pp.10624-10637. ⟨10.1074/jbc.RA120.013906⟩ J Biol Chem |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1074/jbc.RA120.013906⟩ |
| Popis: | International audience; Picornaviral RNA-dependent RNA polymerases (RdRPs) have low replication fidelity that is essential for viral fitness and evolution. Their global fold consists of the classical "cupped right hand" structure with palm, fingers, and thumb domains, and these RdRPs also possess a unique contact between the fingers and thumb domains. This interaction restricts movements of the fingers, and RdRPs use a subtle conformational change within the palm domain to close their active sites for catalysis. We have previously shown that this core RdRP structure and mechanism provide a platform for polymerases to fine-tune replication rates and fidelity to optimize virus fitness. Here, we further elucidated the structural basis for differences in replication rates and fidelity among different viruses by generating chimeric RdRPs from poliovirus and coxsackievirus B3. We designed these chimeric polymerases by exchanging the fingers, pinky finger, or thumb domains. The results of biochemical, rapid-quench, and stopped-flow assays revealed that differences in biochemical activity map to individual modular domains of this polymerase. We found that the pinky finger subdomain is a major regulator of initiation and that the palm domain is the major determinant of catalytic rate and nucleotide discrimination. We further noted that thumb domain interactions with product RNA regulate translocation and that the palm and thumb domains coordinately control elongation complex stability. Several RdRP chimeras supported the growth of infectious poliovirus, providing insights into enterovirus species-specific protein-protein interactions required for virus replication. |
| Databáze: | OpenAIRE |
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