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Furin cleavage of SARS-CoV-2 Spike promotes but is not essential for infection and cell-cell fusion

Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) infects cells by binding to the host cell receptor ACE2 and undergoing virus-host membrane fusion. Fusion is triggered by the protease TMPRSS2, which processes the viral Spike (S) protein to reveal the fusion peptide. SARS-CoV-2 has evolve...

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Autores principales: Papa, Guido, Mallery, Donna L., Albecka, Anna, Welch, Lawrence G., Cattin-Ortolá, Jérôme, Luptak, Jakub, Paul, David, McMahon, Harvey T., Goodfellow, Ian G., Carter, Andrew, Munro, Sean, James, Leo C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7861537/
https://www.ncbi.nlm.nih.gov/pubmed/33493182
http://dx.doi.org/10.1371/journal.ppat.1009246
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author Papa, Guido
Mallery, Donna L.
Albecka, Anna
Welch, Lawrence G.
Cattin-Ortolá, Jérôme
Luptak, Jakub
Paul, David
McMahon, Harvey T.
Goodfellow, Ian G.
Carter, Andrew
Munro, Sean
James, Leo C.
author_facet Papa, Guido
Mallery, Donna L.
Albecka, Anna
Welch, Lawrence G.
Cattin-Ortolá, Jérôme
Luptak, Jakub
Paul, David
McMahon, Harvey T.
Goodfellow, Ian G.
Carter, Andrew
Munro, Sean
James, Leo C.
author_sort Papa, Guido
collection PubMed
description Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) infects cells by binding to the host cell receptor ACE2 and undergoing virus-host membrane fusion. Fusion is triggered by the protease TMPRSS2, which processes the viral Spike (S) protein to reveal the fusion peptide. SARS-CoV-2 has evolved a multibasic site at the S1-S2 boundary, which is thought to be cleaved by furin in order to prime S protein for TMPRSS2 processing. Here we show that CRISPR-Cas9 knockout of furin reduces, but does not prevent, the production of infectious SARS-CoV-2 virus. Comparing S processing in furin knockout cells to multibasic site mutants reveals that while loss of furin substantially reduces S1-S2 cleavage it does not prevent it. SARS-CoV-2 S protein also mediates cell-cell fusion, potentially allowing virus to spread virion-independently. We show that loss of furin in either donor or acceptor cells reduces, but does not prevent, TMPRSS2-dependent cell-cell fusion, unlike mutation of the multibasic site that completely prevents syncytia formation. Our results show that while furin promotes both SARS-CoV-2 infectivity and cell-cell spread it is not essential, suggesting furin inhibitors may reduce but not abolish viral spread.
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spelling pubmed-78615372021-02-12 Furin cleavage of SARS-CoV-2 Spike promotes but is not essential for infection and cell-cell fusion Papa, Guido Mallery, Donna L. Albecka, Anna Welch, Lawrence G. Cattin-Ortolá, Jérôme Luptak, Jakub Paul, David McMahon, Harvey T. Goodfellow, Ian G. Carter, Andrew Munro, Sean James, Leo C. PLoS Pathog Research Article Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) infects cells by binding to the host cell receptor ACE2 and undergoing virus-host membrane fusion. Fusion is triggered by the protease TMPRSS2, which processes the viral Spike (S) protein to reveal the fusion peptide. SARS-CoV-2 has evolved a multibasic site at the S1-S2 boundary, which is thought to be cleaved by furin in order to prime S protein for TMPRSS2 processing. Here we show that CRISPR-Cas9 knockout of furin reduces, but does not prevent, the production of infectious SARS-CoV-2 virus. Comparing S processing in furin knockout cells to multibasic site mutants reveals that while loss of furin substantially reduces S1-S2 cleavage it does not prevent it. SARS-CoV-2 S protein also mediates cell-cell fusion, potentially allowing virus to spread virion-independently. We show that loss of furin in either donor or acceptor cells reduces, but does not prevent, TMPRSS2-dependent cell-cell fusion, unlike mutation of the multibasic site that completely prevents syncytia formation. Our results show that while furin promotes both SARS-CoV-2 infectivity and cell-cell spread it is not essential, suggesting furin inhibitors may reduce but not abolish viral spread. Public Library of Science 2021-01-25 /pmc/articles/PMC7861537/ /pubmed/33493182 http://dx.doi.org/10.1371/journal.ppat.1009246 Text en © 2021 Papa et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://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
Papa, Guido
Mallery, Donna L.
Albecka, Anna
Welch, Lawrence G.
Cattin-Ortolá, Jérôme
Luptak, Jakub
Paul, David
McMahon, Harvey T.
Goodfellow, Ian G.
Carter, Andrew
Munro, Sean
James, Leo C.
Furin cleavage of SARS-CoV-2 Spike promotes but is not essential for infection and cell-cell fusion
title Furin cleavage of SARS-CoV-2 Spike promotes but is not essential for infection and cell-cell fusion
title_full Furin cleavage of SARS-CoV-2 Spike promotes but is not essential for infection and cell-cell fusion
title_fullStr Furin cleavage of SARS-CoV-2 Spike promotes but is not essential for infection and cell-cell fusion
title_full_unstemmed Furin cleavage of SARS-CoV-2 Spike promotes but is not essential for infection and cell-cell fusion
title_short Furin cleavage of SARS-CoV-2 Spike promotes but is not essential for infection and cell-cell fusion
title_sort furin cleavage of sars-cov-2 spike promotes but is not essential for infection and cell-cell fusion
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7861537/
https://www.ncbi.nlm.nih.gov/pubmed/33493182
http://dx.doi.org/10.1371/journal.ppat.1009246
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