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Structural and functional characterization of the Sin Nombre virus L protein

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 counte...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10406178/
https://www.ncbi.nlm.nih.gov/pubmed/37549153
http://dx.doi.org/10.1371/journal.ppat.1011533
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author 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
author_facet 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
author_sort Meier, Kristina
collection PubMed
description 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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spelling pubmed-104061782023-08-08 Structural and functional characterization of the Sin Nombre virus L protein 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 PLoS Pathog Research Article 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. Public Library of Science 2023-08-07 /pmc/articles/PMC10406178/ /pubmed/37549153 http://dx.doi.org/10.1371/journal.ppat.1011533 Text en © 2023 Meier et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
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
Structural and functional characterization of the Sin Nombre virus L protein
title Structural and functional characterization of the Sin Nombre virus L protein
title_full Structural and functional characterization of the Sin Nombre virus L protein
title_fullStr Structural and functional characterization of the Sin Nombre virus L protein
title_full_unstemmed Structural and functional characterization of the Sin Nombre virus L protein
title_short Structural and functional characterization of the Sin Nombre virus L protein
title_sort structural and functional characterization of the sin nombre virus l protein
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10406178/
https://www.ncbi.nlm.nih.gov/pubmed/37549153
http://dx.doi.org/10.1371/journal.ppat.1011533
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