The role of furin cleavage site in SARS-CoV-2 spike protein-mediated membrane fusion in the presence or absence of trypsin

Xia, Shuai; Lan, Qiaoshuai; Su, Shan; Wang, Xinling; Xu, Wei; Liu, Zezhong; Zhu, Yun; Wang, Qian; Lu, Lu; Jiang, Shibo

doi:10.1038/s41392-020-0184-0

Cited by 319 publications

(365 citation statements)

References 5 publications

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“…Furin is widely assumed to be the protease responsible for cleavage of the CoV-2 S protein multibasic site. However, most studies have relied on indirect methodologies such as small molecule inhibition or mRNA depletion (Hoffmann et al, 2020b;Shang et al, 2020;Xia et al, 2020). We decided to investigate the importance of furin directly by generating a 293T CRISPR-derived knockout cell line (herein named 293T-∆FURIN).…”

Section: Furin Protease Enhances Cov-2 S Protein Cleavage But It Is Nmentioning

confidence: 99%

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

Papa

Mallery

Albecka

et al. 2020

Preprint

147

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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 will not prevent viral spread.

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Section: Furin Protease Enhances Cov-2 S Protein Cleavage But It Is Nmentioning

confidence: 99%

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

Papa

Mallery

Albecka

et al. 2020

Preprint

147

View full text Add to dashboard Cite

show abstract

“…The fusion capacity of the S protein is a leading indicator of the infectivity of the virus. S protein consists of two functional subunits, receptor-binding (S 1 ) and fusion the viral and cellular membrane (S 2 ) 2,3 . S 1 comprises the receptor-binding domain (RBD) that recognizes angiotensin-converting enzyme-2 (ACE2) as its receptor 4,5 , S 1 contributes to the stabilization of the prefusion state of the membrane-anchored S 2 subunit, that contains the fusion machinery.…”

Section: Introductionmentioning

confidence: 99%

“…The priming process is ensured by different host cell proteases depending on the S1/S2 sequence cleavage site. In the case of the SARS-CoV-2, S protein contains a canonical furin-like cleavage site: 681-PRRAR↓SV-688, which is absent in SARS-CoV and others SARS-related coronaviruses 2,3,6,7 , and it may provide a gain of function for e cient spreading in the human population compared to other lineages of beta-coronaviruses 6 .…”

Section: Introductionmentioning

confidence: 99%

Structural insights into spike protein and its natural variants of SARS-CoV-2 found on Mexican population.

Sixto-López

Bello

Correa-Basurto

et al. 2020

Preprint

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The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a newly emerged coronavirus responsible for COVID-19; it becomes a pandemic since March 2020. To date, there are described three lineages of SARS-CoV-2 circulating worldwide, in Mexican population are found two of them, within this, we observed three variants of Spike (S) protein located at H49Y, D614G, and T573I. In order to understand if these mutations could affect the structural behavior of S protein of SARS-CoV-2, as well as the binding with three experimental describe inhibitors (Cepharanthine, Nelfinavir, and Hydroxychloroquine), molecular dynamic simulation and molecular docking were employed. It was found that in spite, these punctual mutations affect considerably the structural behavior of the S Protein, which also affect the binding of the inhibitors into their respective binding site. Thus, further experimental studies need to be done in order to explore if these affectations have an impact on drug-S protein binding and the possible clinical effect.

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“…17 However, a recent study suggested the furin cleavage motif is not essential for the fusion activity of SARS-CoV-2 spike protein in the environment where serine proteases are present, such as the human airway. 18 Given the controversies, we sought to evaluate the role of the PRRA motif for SARS-CoV-2 replication in human airway organoids. We obtained a SARS-CoV-2 variant with PRRA motif deletion through plaque purification and compared the replication kinetics of wildtype and deletion mutant virus.…”

Section: One Of the Notable Features Of Sars-cov-2 Is The Presence Ofmentioning

confidence: 99%

“…However, these meticulously-conducted studies reached conflicting conclusions. 16,18 The inspection in human airway organoids indicates furin cleavage is dispensable for the infectivity and replication capacity of SARS-CoV-2. Furthermore, viral genome region harboring the furin cleavage motif is very stable during SARS-CoV-2 replication in human airway organoids, in contrast to the rapid acquisition of the deletion mutation in Vero E6 cells.…”

Section: One Of the Notable Features Of Sars-cov-2 Is The Presence Ofmentioning

confidence: 99%

Human airway organoids model SARS-CoV-2 high infectiousness and evasion of interferon response

Zhou¹,

Chiu²,

Li³

et al. 2020

Preprint

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SARS-CoV-2 is more infectious and transmissible in humans than SARS-CoV, despite the genetic relatedness and sharing the same cellular receptor. We sought to assess whether human airway organoids can model SARS-CoV-2 infection in the human airway and elucidate the cellular basis underlying its higher transmissibility. We demonstrate that SARS-CoV-2 can establish a productive infection in human airway organoids, in which ciliated cell and basal cell are infected. Wildtype SARS-CoV-2 carrying a furin cleavage motif exhibits comparable replication kinetics to a mutant virus without the motif. Human airway organoids sustain higher replication of SARS-CoV-2 than SARS-CoV, whereas interferon response is more potently induced in the latter than the former. Overall, human airway organoids can model SARS-CoV-2 infection and recapitulate the disposable role of furin cleavage motif for virus transmission in humans. SARS-CoV-2 stealth growth and evasion of interferon response may underlie pre-symptomatic virus shedding in COVID-19 patients, leading to its high infectiousness and transmissibility.

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The role of furin cleavage site in SARS-CoV-2 spike protein-mediated membrane fusion in the presence or absence of trypsin

Cited by 319 publications

References 5 publications

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

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

Structural insights into spike protein and its natural variants of SARS-CoV-2 found on Mexican population.

Human airway organoids model SARS-CoV-2 high infectiousness and evasion of interferon response

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