Nsp1 proteins of human coronaviruses HCoVOC43 and SARS-CoV2 inhibit stress granule formation

Stacia M. Dolliver; Mariel Kleer; Maxwell P. Bui-Marinos; Shan Ying; Jennifer A. Corcoran; Denys A. Khaperskyy

doi:10.1371/journal.ppat.1011041

Nsp1 proteins of human coronaviruses HCoVOC43 and SARS-CoV2 inhibit stress granule formation

Stacia M. Dolliver, Mariel Kleer, Maxwell P. Bui-Marinos, Shan Ying, Jennifer A. Corcoran, Denys A. Khaperskyy

Medicine

Résultat de recherche: Article › examen par les pairs

22 Citations (Scopus)

Résumé

Stress granules (SGs) are cytoplasmic condensates that often form as part of the cellular antiviral response. Despite the growing interest in understanding the interplay between SGs and other biological condensates and viral replication, the role of SG formation during coronavirus infection remains poorly understood. Several proteins from different coronaviruses have been shown to suppress SG formation upon overexpression, but there are only a handful of studies analyzing SG formation in coronavirus-infected cells. To better understand SG inhibition by coronaviruses, we analyzed SG formation during infection with the human common cold coronavirus OC43 (HCoV-OC43) and the pandemic SARS-CoV2. We did not observe SG induction in infected cells and both viruses inhibited eukaryotic translation initiation factor 2α (eIF2α) phosphorylation and SG formation induced by exogenous stress. Furthermore, in SARS-CoV2 infected cells we observed a sharp decrease in the levels of SG-nucleating protein G3BP1. Ectopic overexpression of nucleocapsid (N) and nonstructural protein 1 (Nsp1) from both HCoV-OC43 and SARS-CoV2 inhibited SG formation. The Nsp1 proteins of both viruses inhibited arsenite-induced eIF2α phosphorylation, and the Nsp1 of SARS-CoV2 alone was sufficient to cause a decrease in G3BP1 levels. This phenotype was dependent on the depletion of cytoplasmic mRNA mediated by Nsp1 and associated with nuclear accumulation of the SG-nucleating protein TIAR. To test the role of G3BP1 in coronavirus replication, we infected cells overexpressing EGFP-tagged G3BP1 with HCoV-OC43 and observed a significant decrease in virus replication compared to control cells expressing EGFP. The antiviral role of G3BP1 and the existence of multiple SG suppression mechanisms that are conserved between HCoV-OC43 and SARS-CoV2 suggest that SG formation may represent an important antiviral host defense that coronaviruses target to ensure efficient replication.

Langue d'origine	English
Numéro d'article	e1011041
Journal	PLoS Pathogens
Volume	18
Numéro de publication	12
DOI	https://doi.org/10.1371/journal.ppat.1011041
Statut de publication	Published - déc. 2022

Note bibliographique

Funding Information:
This work was supported by Canadian Institutes for Health Research (CIHR) Project Grant PJT-175130 and Research Nova Scotia Grant RNS-NHIG-2020-1383 (to D.K.). This study was also supported in part by operating funds awarded to JAC from the Canadian Institutes for Health Research (CIHR): a COVID rapid response operating grant (#177704) and an operating grant (#175622) the Coronavirus Variants Rapid Response Network (CoVaRR-Net), of which JAC is a member. MK was supported by a Cumming School of Medicine graduate training award, a Canadian Institutes for Health Research (CIHR) CGS-M scholarship, and a CIHR doctoral award. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We would like to thank Dr. Arinjay Banerjee (Vaccine and Infectious Disease Organization (VIDO), Dr. Karen Mossman (University of McMaster) and Dr. Samira Mubareka (University of Toronto) for the SARS-CoV-2 isolate. We also thank Dr. Craig McCormick (Dalhousie University) for providing NL63 cDNA and Dr. Marta Gaglia (University of Wisconsin-Madison) for providing SARS Nsp1 plasmid. We also thank Dalhousie CORES Flow Cytometry Facility for assistance in generating stable cell lines expressing EGFP-G3BP1 and EGFP. Finally, we thank members of Khaperskyy and Corcoran labs for helpful discussions about experimental design and their critical input on the draft manuscript.

Publisher Copyright:
Copyright: © 2022 Dolliver et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

ASJC Scopus Subject Areas

Parasitology
Microbiology
Immunology
Molecular Biology
Genetics
Virology

PubMed: MeSH publication types

Journal Article

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10.1371/journal.ppat.1011041

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title = "Nsp1 proteins of human coronaviruses HCoVOC43 and SARS-CoV2 inhibit stress granule formation",

abstract = "Stress granules (SGs) are cytoplasmic condensates that often form as part of the cellular antiviral response. Despite the growing interest in understanding the interplay between SGs and other biological condensates and viral replication, the role of SG formation during coronavirus infection remains poorly understood. Several proteins from different coronaviruses have been shown to suppress SG formation upon overexpression, but there are only a handful of studies analyzing SG formation in coronavirus-infected cells. To better understand SG inhibition by coronaviruses, we analyzed SG formation during infection with the human common cold coronavirus OC43 (HCoV-OC43) and the pandemic SARS-CoV2. We did not observe SG induction in infected cells and both viruses inhibited eukaryotic translation initiation factor 2α (eIF2α) phosphorylation and SG formation induced by exogenous stress. Furthermore, in SARS-CoV2 infected cells we observed a sharp decrease in the levels of SG-nucleating protein G3BP1. Ectopic overexpression of nucleocapsid (N) and nonstructural protein 1 (Nsp1) from both HCoV-OC43 and SARS-CoV2 inhibited SG formation. The Nsp1 proteins of both viruses inhibited arsenite-induced eIF2α phosphorylation, and the Nsp1 of SARS-CoV2 alone was sufficient to cause a decrease in G3BP1 levels. This phenotype was dependent on the depletion of cytoplasmic mRNA mediated by Nsp1 and associated with nuclear accumulation of the SG-nucleating protein TIAR. To test the role of G3BP1 in coronavirus replication, we infected cells overexpressing EGFP-tagged G3BP1 with HCoV-OC43 and observed a significant decrease in virus replication compared to control cells expressing EGFP. The antiviral role of G3BP1 and the existence of multiple SG suppression mechanisms that are conserved between HCoV-OC43 and SARS-CoV2 suggest that SG formation may represent an important antiviral host defense that coronaviruses target to ensure efficient replication.",

author = "Dolliver, {Stacia M.} and Mariel Kleer and Bui-Marinos, {Maxwell P.} and Shan Ying and Corcoran, {Jennifer A.} and Khaperskyy, {Denys A.}",

note = "Funding Information: This work was supported by Canadian Institutes for Health Research (CIHR) Project Grant PJT-175130 and Research Nova Scotia Grant RNS-NHIG-2020-1383 (to D.K.). This study was also supported in part by operating funds awarded to JAC from the Canadian Institutes for Health Research (CIHR): a COVID rapid response operating grant (#177704) and an operating grant (#175622) the Coronavirus Variants Rapid Response Network (CoVaRR-Net), of which JAC is a member. MK was supported by a Cumming School of Medicine graduate training award, a Canadian Institutes for Health Research (CIHR) CGS-M scholarship, and a CIHR doctoral award. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We would like to thank Dr. Arinjay Banerjee (Vaccine and Infectious Disease Organization (VIDO), Dr. Karen Mossman (University of McMaster) and Dr. Samira Mubareka (University of Toronto) for the SARS-CoV-2 isolate. We also thank Dr. Craig McCormick (Dalhousie University) for providing NL63 cDNA and Dr. Marta Gaglia (University of Wisconsin-Madison) for providing SARS Nsp1 plasmid. We also thank Dalhousie CORES Flow Cytometry Facility for assistance in generating stable cell lines expressing EGFP-G3BP1 and EGFP. Finally, we thank members of Khaperskyy and Corcoran labs for helpful discussions about experimental design and their critical input on the draft manuscript. Publisher Copyright: Copyright: {\textcopyright} 2022 Dolliver et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

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AU - Dolliver, Stacia M.

AU - Kleer, Mariel

AU - Bui-Marinos, Maxwell P.

AU - Ying, Shan

AU - Corcoran, Jennifer A.

AU - Khaperskyy, Denys A.

N1 - Funding Information: This work was supported by Canadian Institutes for Health Research (CIHR) Project Grant PJT-175130 and Research Nova Scotia Grant RNS-NHIG-2020-1383 (to D.K.). This study was also supported in part by operating funds awarded to JAC from the Canadian Institutes for Health Research (CIHR): a COVID rapid response operating grant (#177704) and an operating grant (#175622) the Coronavirus Variants Rapid Response Network (CoVaRR-Net), of which JAC is a member. MK was supported by a Cumming School of Medicine graduate training award, a Canadian Institutes for Health Research (CIHR) CGS-M scholarship, and a CIHR doctoral award. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We would like to thank Dr. Arinjay Banerjee (Vaccine and Infectious Disease Organization (VIDO), Dr. Karen Mossman (University of McMaster) and Dr. Samira Mubareka (University of Toronto) for the SARS-CoV-2 isolate. We also thank Dr. Craig McCormick (Dalhousie University) for providing NL63 cDNA and Dr. Marta Gaglia (University of Wisconsin-Madison) for providing SARS Nsp1 plasmid. We also thank Dalhousie CORES Flow Cytometry Facility for assistance in generating stable cell lines expressing EGFP-G3BP1 and EGFP. Finally, we thank members of Khaperskyy and Corcoran labs for helpful discussions about experimental design and their critical input on the draft manuscript. Publisher Copyright: Copyright: © 2022 Dolliver et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PY - 2022/12

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N2 - Stress granules (SGs) are cytoplasmic condensates that often form as part of the cellular antiviral response. Despite the growing interest in understanding the interplay between SGs and other biological condensates and viral replication, the role of SG formation during coronavirus infection remains poorly understood. Several proteins from different coronaviruses have been shown to suppress SG formation upon overexpression, but there are only a handful of studies analyzing SG formation in coronavirus-infected cells. To better understand SG inhibition by coronaviruses, we analyzed SG formation during infection with the human common cold coronavirus OC43 (HCoV-OC43) and the pandemic SARS-CoV2. We did not observe SG induction in infected cells and both viruses inhibited eukaryotic translation initiation factor 2α (eIF2α) phosphorylation and SG formation induced by exogenous stress. Furthermore, in SARS-CoV2 infected cells we observed a sharp decrease in the levels of SG-nucleating protein G3BP1. Ectopic overexpression of nucleocapsid (N) and nonstructural protein 1 (Nsp1) from both HCoV-OC43 and SARS-CoV2 inhibited SG formation. The Nsp1 proteins of both viruses inhibited arsenite-induced eIF2α phosphorylation, and the Nsp1 of SARS-CoV2 alone was sufficient to cause a decrease in G3BP1 levels. This phenotype was dependent on the depletion of cytoplasmic mRNA mediated by Nsp1 and associated with nuclear accumulation of the SG-nucleating protein TIAR. To test the role of G3BP1 in coronavirus replication, we infected cells overexpressing EGFP-tagged G3BP1 with HCoV-OC43 and observed a significant decrease in virus replication compared to control cells expressing EGFP. The antiviral role of G3BP1 and the existence of multiple SG suppression mechanisms that are conserved between HCoV-OC43 and SARS-CoV2 suggest that SG formation may represent an important antiviral host defense that coronaviruses target to ensure efficient replication.

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Nsp1 proteins of human coronaviruses HCoVOC43 and SARS-CoV2 inhibit stress granule formation

Résumé

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PubMed: MeSH publication types

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