The furin cleavage site in the SARS-CoV-2 spike protein is required for transmission in ferrets.
Animals
COVID-19
/ transmission
Cathepsins
/ metabolism
Chlorocebus aethiops
Endosomes
/ metabolism
Epithelial Cells
Ferrets
Furin
/ metabolism
Humans
Immune Evasion
Membrane Proteins
/ metabolism
RNA-Binding Proteins
/ metabolism
Respiratory System
/ cytology
SARS-CoV-2
/ physiology
Serine Endopeptidases
/ metabolism
Spike Glycoprotein, Coronavirus
/ chemistry
Vero Cells
Viral Genome Packaging
Virus Internalization
Virus Replication
Virus Shedding
Journal
Nature microbiology
ISSN: 2058-5276
Titre abrégé: Nat Microbiol
Pays: England
ID NLM: 101674869
Informations de publication
Date de publication:
07 2021
07 2021
Historique:
received:
13
10
2020
accepted:
08
04
2021
pubmed:
29
4
2021
medline:
9
7
2021
entrez:
28
4
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 33907312
doi: 10.1038/s41564-021-00908-w
pii: 10.1038/s41564-021-00908-w
doi:
Substances chimiques
IFITM3 protein, human
0
Membrane Proteins
0
RNA-Binding Proteins
0
Spike Glycoprotein, Coronavirus
0
spike protein, SARS-CoV-2
0
Cathepsins
EC 3.4.-
Serine Endopeptidases
EC 3.4.21.-
TMPRSS2 protein, human
EC 3.4.21.-
Furin
EC 3.4.21.75
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
899-909Subventions
Organisme : RCUK | Biotechnology and Biological Sciences Research Council (BBSRC)
ID : BB/R013071/1
Organisme : RCUK | Biotechnology and Biological Sciences Research Council (BBSRC)
ID : BB/R007292/1
Organisme : RCUK | Biotechnology and Biological Sciences Research Council (BBSRC)
ID : BB/K002465/1
Organisme : RCUK | Biotechnology and Biological Sciences Research Council (BBSRC)
ID : DTP studentship
Organisme : RCUK | Biotechnology and Biological Sciences Research Council (BBSRC)
ID : BB/M02542X/1
Organisme : RCUK | Biotechnology and Biological Sciences Research Council (BBSRC)
ID : BB/S008292/1
Organisme : Wellcome Trust
ID : 205100
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 200187
Pays : United Kingdom
Organisme : Wellcome Trust (Wellcome)
ID : studentship
Organisme : Wellcome Trust (Wellcome)
ID : 216353/Z/19/Z
Organisme : RCUK | Medical Research Council (MRC)
ID : DTP studentship
Organisme : RCUK | Medical Research Council (MRC)
ID : MR/R020566/1
Organisme : Imperial College London
ID : President's Scholarship
Organisme : Kings College London
ID : RE/18/2/34213
Organisme : Wellcome Trust
ID : 205100
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 200187
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 200187
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 205100
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 205100
Pays : United Kingdom
Organisme : FDA HHS
ID : HHSF223201510104C
Pays : United States
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