Abstract
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.
【초록키워드】 SARS-CoV-2, Antiviral, furin, spike glycoprotein, lung, SARS-CoV-2 virus, Proteins, Replication, SARS-CoV-2 transmission, ferret, pseudotype, furin cleavage site, Patient, virus entry, Deletion, cleavage, epithelial cells, membrane fusion, mutant, expression, Live virus, Frequency, S1/S2 cleavage site, epithelial cell, Vero E6, growth, SARS-CoV-2 entry, S1/S2, SARS-CoV-2 sequence, titre, cleavage site, wild-type virus, insertion, protease TMPRSS2, cell line, selective, IFITM, primary human airway, Cell, lung cell, transmitted, impair, cleaved, lentiviral, polybasic, postmortem tissue, spike variant, 【제목키워드】 Transmission, Spike protein, ferret, furin cleavage site, required, the SARS-CoV-2,