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The furin cleavage site in the SARS-CoV-2 spike protein is required for transmission in ferrets.
Nat Microbiol. 2021 Jul; 6(7):899-909.NM

Abstract

SARS-CoV-2 entry requires sequential cleavage of the spike glycoprotein at the S1/S2 and the S2' 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.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Peacock TP
Department of Infectious Diseases, Imperial College London, London, UK.
Goldhill DH
Department of Infectious Diseases, Imperial College London, London, UK.
Zhou J
Department of Infectious Diseases, Imperial College London, London, UK.
Baillon L
Department of Infectious Diseases, Imperial College London, London, UK.
Frise R
Department of Infectious Diseases, Imperial College London, London, UK.
Swann OC
Department of Infectious Diseases, Imperial College London, London, UK.
Kugathasan R
Department of Infectious Diseases, Imperial College London, London, UK.
Penn R
Department of Infectious Diseases, Imperial College London, London, UK.
Brown JC
Department of Infectious Diseases, Imperial College London, London, UK.
Sanchez-David RY
Department of Infectious Diseases, Imperial College London, London, UK.
Braga L
British Hearth Foundation Centre of Research Excellence, School of Cardiovascular Medicine and Sciences, King's College London, London, UK.
Williamson MK
School of Cellular and Molecular Medicine, Faculty of Life Sciences, University of Bristol, Bristol, UK.
Hassard JA
Department of Infectious Diseases, Imperial College London, London, UK.
Staller E
Department of Infectious Diseases, Imperial College London, London, UK.
Hanley B
Department of Cellular Pathology, Northwest London Pathology, Imperial College London NHS Trust, London, UK.
Osborn M
Department of Cellular Pathology, Northwest London Pathology, Imperial College London NHS Trust, London, UK.
Giacca M
British Hearth Foundation Centre of Research Excellence, School of Cardiovascular Medicine and Sciences, King's College London, London, UK.
Davidson AD
School of Cellular and Molecular Medicine, Faculty of Life Sciences, University of Bristol, Bristol, UK.
Matthews DA
School of Cellular and Molecular Medicine, Faculty of Life Sciences, University of Bristol, Bristol, UK.
Barclay WS
Department of Infectious Diseases, Imperial College London, London, UK. w.barclay@imperial.ac.uk.

MeSH

AnimalsCOVID-19CathepsinsChlorocebus aethiopsEndosomesEpithelial CellsFerretsFurinHumansImmune EvasionMembrane ProteinsRNA-Binding ProteinsRespiratory SystemSARS-CoV-2Serine EndopeptidasesSpike Glycoprotein, CoronavirusVero CellsViral Genome PackagingVirus InternalizationVirus ReplicationVirus Shedding

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

33907312

Citation

Peacock, Thomas P., et al. "The Furin Cleavage Site in the SARS-CoV-2 Spike Protein Is Required for Transmission in Ferrets." Nature Microbiology, vol. 6, no. 7, 2021, pp. 899-909.
Peacock TP, Goldhill DH, Zhou J, et al. The furin cleavage site in the SARS-CoV-2 spike protein is required for transmission in ferrets. Nat Microbiol. 2021;6(7):899-909.
Peacock, T. P., Goldhill, D. H., Zhou, J., Baillon, L., Frise, R., Swann, O. C., Kugathasan, R., Penn, R., Brown, J. C., Sanchez-David, R. Y., Braga, L., Williamson, M. K., Hassard, J. A., Staller, E., Hanley, B., Osborn, M., Giacca, M., Davidson, A. D., Matthews, D. A., & Barclay, W. S. (2021). The furin cleavage site in the SARS-CoV-2 spike protein is required for transmission in ferrets. Nature Microbiology, 6(7), 899-909. https://doi.org/10.1038/s41564-021-00908-w
Peacock TP, et al. The Furin Cleavage Site in the SARS-CoV-2 Spike Protein Is Required for Transmission in Ferrets. Nat Microbiol. 2021;6(7):899-909. PubMed PMID: 33907312.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The furin cleavage site in the SARS-CoV-2 spike protein is required for transmission in ferrets. AU - Peacock,Thomas P, AU - Goldhill,Daniel H, AU - Zhou,Jie, AU - Baillon,Laury, AU - Frise,Rebecca, AU - Swann,Olivia C, AU - Kugathasan,Ruthiran, AU - Penn,Rebecca, AU - Brown,Jonathan C, AU - Sanchez-David,Raul Y, AU - Braga,Luca, AU - Williamson,Maia Kavanagh, AU - Hassard,Jack A, AU - Staller,Ecco, AU - Hanley,Brian, AU - Osborn,Michael, AU - Giacca,Mauro, AU - Davidson,Andrew D, AU - Matthews,David A, AU - Barclay,Wendy S, Y1 - 2021/04/27/ PY - 2020/10/13/received PY - 2021/4/8/accepted PY - 2021/4/29/pubmed PY - 2021/7/9/medline PY - 2021/4/28/entrez SP - 899 EP - 909 JF - Nature microbiology JO - Nat Microbiol VL - 6 IS - 7 N2 - SARS-CoV-2 entry requires sequential cleavage of the spike glycoprotein at the S1/S2 and the S2' 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. SN - 2058-5276 UR - https://www.unboundmedicine.com/medline/citation/33907312/The_furin_cleavage_site_in_the_SARS_CoV_2_spike_protein_is_required_for_transmission_in_ferrets_ DB - PRIME DP - Unbound Medicine ER -