Structural and functional characterization of the Sin Nombre virus L protein
In: ISSN: 1553-7366, 2023
Online
academicJournal
Zugriff:
International audience ; The Bunyavirales order is a large and diverse group of segmented negative-strand RNA viruses. Several virus families within this order contain important human pathogens, including Sin Nombre virus (SNV) of the Hantaviridae. Despite the high epidemic potential of bunyaviruses, specific medical countermeasures such as vaccines or antivirals are missing. The multifunctional ~250 kDa L protein of hantaviruses, amongst other functional domains, harbors the RNA-dependent RNA polymerase (RdRp) and an endonuclease and catalyzes transcription as well as replication of the viral RNA genome, making it a promising therapeutic target. The development of inhibitors targeting these key processes requires a profound understanding of the catalytic mechanisms. Here, we established expression and purification protocols of the full-length SNV L protein bearing the endonuclease mutation K124A. We applied different biochemical in vitro assays to provide an extensive characterization of the different enzymatic functions as well as the capacity of the hantavirus L protein to interact with the viral RNA. By using single-particle cryo-EM, we obtained a 3D model including the L protein core region containing the RdRp, in complex with the 5′ promoter RNA. This first high-resolution model of a New World hantavirus L protein shows striking similarity to related bunyavirus L proteins. The interaction of the L protein with the 5′ RNA observed in the structural model confirms our hypothesis of protein-RNA binding based on our biochemical data. Taken together, this study provides an excellent basis for future structural and functional studies on the hantavirus L protein and for the development of antiviral compounds.
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Structural and functional characterization of the Sin Nombre virus L protein
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Autor/in / Beteiligte Person: | Meier, Kristina ; Thorkelsson, Sigurdur, R ; Durieux Trouilleton, Quentin ; Vogel, Dominik ; Yu, Dingquan ; Kosinski, Jan ; Cusack, Stephen ; Malet, Hélène ; Grünewald, Kay ; Quemin, Emmanuelle, R J ; Rosenthal, Maria ; Bernhard Nocht Institute for Tropical Medicine - Bernhard-Nocht-Institut für Tropenmedizin Hamburg, Germany (BNITM) ; Leibniz-Institut für Virologie Hamburg (LIV) ; Groupe Imagerie intégrée de la réponse au stress / Integrated Imaging of Stress Response Group (IBS-I2SR) ; 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)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG) ; Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA) ; European Molecular Biology Laboratory Grenoble (EMBL) ; Institut universitaire de France (IUF) ; Ministère de l'Education nationale, de l’Enseignement supérieur et de la Recherche (M.E.N.E.S.R.) ; Universität Hamburg (UHH) ; Institut de Biologie Intégrative de la Cellule (I2BC) ; Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS) |
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Zeitschrift: | ISSN: 1553-7366, 2023 |
Veröffentlichung: | HAL CCSD ; Public Library of Science, 2023 |
Medientyp: | academicJournal |
DOI: | 10.1371/journal.ppat.1011533 |
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