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nature portfolio permissions reprints privacy policy cell-culture-adapted sars-cov-2 virus rna extraction kits advertising early development rabbit anti-sars-cov-2 nucleocapsid sars-cov s1/s2 junction tools regulate sars-cov-2 entry nature https china attenuated sars-cov-2 variants sars-cov-2prra variants emergent sars-cov-2 lineage emergent-sars-cov-2-lineage sars-cov-2 spike protein infra-red secondary antibodies nature 579 nature 426 nature efficient serine-protease-dependent entry c-terminal flag-tag sars-cov-2 spike variants closely related sars-cov research excellence grant wider sars-related coronaviruses sequential proteolytic cleavage thermo fisher scientific mda-5-dependent interferon response codon-optimized spike proteins rabbit anti-fragilis/ifitm3 sars-cov-2 spike glycoprotein blocking sars-cov-2 replication s1/s2 cleavage site mixed sars-cov-2 population sars-cov-2 animal models20 multiple sars-cov-2 variants sars-cov-2 strain england/2/2020 sars-cov-2 spike mutants endosomal/endolysosomal ifitm proteins sars-cov-2 virus lacking sars-cov-2 overlay medium sars-cov wt spike furin-cleavable spike proteins world health organization1 δcs mutant sars-cov-2 s1/s2 cs predicted peer review

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         description:SARS-CoV-2 entry requires sequential cleavage of the spike glycoprotein at the S1/S2 and the S2ʹ cleavage sites to mediate membrane fusion. SARS-CoV-2 has a polybasic insertion (PRRAR) at the S1/S2 cleavage site that can be cleaved by furin. Using lentiviral pseudotypes and a cell-culture-adapted SARS-CoV-2 virus with an S1/S2 deletion, we show that the polybasic insertion endows SARS-CoV-2 with a selective advantage in lung cells and primary human airway epithelial cells, but impairs replication in Vero E6, a cell line used for passaging SARS-CoV-2. Using engineered spike variants and live virus competition assays and by measuring growth kinetics, we find that the selective advantage in lung and primary human airway epithelial cells depends on the expression of the cell surface protease TMPRSS2, which enables endosome-independent virus entry by a route that avoids antiviral IFITM proteins. SARS-CoV-2 virus lacking the S1/S2 furin cleavage site was shed to lower titres from infected ferrets and was not transmitted to cohoused sentinel animals, unlike wild-type virus. Analysis of 100,000 SARS-CoV-2 sequences derived from patients and 24 human postmortem tissues showed low frequencies of naturally occurring mutants that harbour deletions at the polybasic site. Taken together, our findings reveal that the furin cleavage site is an important determinant of SARS-CoV-2 transmission. Efficient transmission of SARS-CoV-2 among infected ferrets requires a functional furin cleavage site in the SARS-CoV-2 spike protein.
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            address:
               name:Department of Infectious Diseases, Imperial College London, London, UK
               type:PostalAddress
            type:Organization
      name:Ruthiran Kugathasan
      affiliation:
            name:Imperial College London
            address:
               name:Department of Infectious Diseases, Imperial College London, London, UK
               type:PostalAddress
            type:Organization
      name:Rebecca Penn
      affiliation:
            name:Imperial College London
            address:
               name:Department of Infectious Diseases, Imperial College London, London, UK
               type:PostalAddress
            type:Organization
      name:Jonathan C. Brown
      affiliation:
            name:Imperial College London
            address:
               name:Department of Infectious Diseases, Imperial College London, London, UK
               type:PostalAddress
            type:Organization
      name:Raul Y. Sanchez-David
      affiliation:
            name:Imperial College London
            address:
               name:Department of Infectious Diseases, Imperial College London, London, UK
               type:PostalAddress
            type:Organization
      name:Luca Braga
      affiliation:
            name:King’s College London
            address:
               name:British Hearth Foundation Centre of Research Excellence, School of Cardiovascular Medicine and Sciences, King’s College London, London, UK
               type:PostalAddress
            type:Organization
      name:Maia Kavanagh Williamson
      affiliation:
            name:University of Bristol
            address:
               name:School of Cellular and Molecular Medicine, Faculty of Life Sciences, University of Bristol, Bristol, UK
               type:PostalAddress
            type:Organization
      name:Jack A. Hassard
      affiliation:
            name:Imperial College London
            address:
               name:Department of Infectious Diseases, Imperial College London, London, UK
               type:PostalAddress
            type:Organization
      name:Ecco Staller
      affiliation:
            name:Imperial College London
            address:
               name:Department of Infectious Diseases, Imperial College London, London, UK
               type:PostalAddress
            type:Organization
      name:Brian Hanley
      affiliation:
            name:Northwest London Pathology, Imperial College London NHS Trust
            address:
               name:Department of Cellular Pathology, Northwest London Pathology, Imperial College London NHS Trust, London, UK
               type:PostalAddress
            type:Organization
      name:Michael Osborn
      affiliation:
            name:Northwest London Pathology, Imperial College London NHS Trust
            address:
               name:Department of Cellular Pathology, Northwest London Pathology, Imperial College London NHS Trust, London, UK
               type:PostalAddress
            type:Organization
      name:Mauro Giacca
      url:http://orcid.org/0000-0003-2927-7225
      affiliation:
            name:King’s College London
            address:
               name:British Hearth Foundation Centre of Research Excellence, School of Cardiovascular Medicine and Sciences, King’s College London, London, UK
               type:PostalAddress
            type:Organization
      name:Andrew D. Davidson
      affiliation:
            name:University of Bristol
            address:
               name:School of Cellular and Molecular Medicine, Faculty of Life Sciences, University of Bristol, Bristol, UK
               type:PostalAddress
            type:Organization
      name:David A. Matthews
      url:http://orcid.org/0000-0003-4611-8795
      affiliation:
            name:University of Bristol
            address:
               name:School of Cellular and Molecular Medicine, Faculty of Life Sciences, University of Bristol, Bristol, UK
               type:PostalAddress
            type:Organization
      name:Wendy S. Barclay
      url:http://orcid.org/0000-0002-3948-0895
      affiliation:
            name:Imperial College London
            address:
               name:Department of Infectious Diseases, Imperial College London, London, UK
               type:PostalAddress
            type:Organization
      email:[email protected]
PostalAddress:
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:British Hearth Foundation Centre of Research Excellence, School of Cardiovascular Medicine and Sciences, King’s College London, London, UK
      name:School of Cellular and Molecular Medicine, Faculty of Life Sciences, University of Bristol, Bristol, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK
      name:Department of Cellular Pathology, Northwest London Pathology, Imperial College London NHS Trust, London, UK
      name:Department of Cellular Pathology, Northwest London Pathology, Imperial College London NHS Trust, London, UK
      name:British Hearth Foundation Centre of Research Excellence, School of Cardiovascular Medicine and Sciences, King’s College London, London, UK
      name:School of Cellular and Molecular Medicine, Faculty of Life Sciences, University of Bristol, Bristol, UK
      name:School of Cellular and Molecular Medicine, Faculty of Life Sciences, University of Bristol, Bristol, UK
      name:Department of Infectious Diseases, Imperial College London, London, UK

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