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SARS-CoV-2 spike engagement of ACE2 primes S2' site cleavage and fusion initiation.
Proc Natl Acad Sci U S A. 2022 01 04; 119(1)PN

Abstract

The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has resulted in tremendous loss worldwide. Although viral spike (S) protein binding of angiotensin-converting enzyme 2 (ACE2) has been established, the functional consequences of the initial receptor binding and the stepwise fusion process are not clear. By utilizing a cell-cell fusion system, in complement with a pseudoviral infection model, we found that the spike engagement of ACE2 primed the generation of S2' fragments in target cells, a key proteolytic event coupled with spike-mediated membrane fusion. Mutagenesis of an S2' cleavage site at the arginine (R) 815, but not an S2 cleavage site at arginine 685, was sufficient to prevent subsequent syncytia formation and infection in a variety of cell lines and primary cells isolated from human ACE2 knock-in mice. The requirement for S2' cleavage at the R815 site was also broadly shared by other SARS-CoV-2 spike variants, such as the Alpha, Beta, and Delta variants of concern. Thus, our study highlights an essential role for host receptor engagement and the key residue of spike for proteolytic activation, and uncovers a targetable mechanism for host cell infection by SARS-CoV-2.

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Authors+Show Affiliations

Yu S
The Center for Microbes, Development and Health, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China. CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
Zheng X
The Center for Microbes, Development and Health, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China. CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
Zhou B
CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
Li J
The Center for Microbes, Development and Health, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China. CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
Chen M
The Center for Microbes, Development and Health, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China. CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
Deng R
The Center for Microbes, Development and Health, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China. CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
Wong G
CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
Lavillette D
CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China; dlaville@ips.ac.cn gxmeng@ips.ac.cn. Pasteurien College, Soochow University, Suzhou, Jiangsu 215006, China. Nanjing Unicorn Academy of Innovation, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Nanjing, Jiangsu 211135, China.
Meng G
The Center for Microbes, Development and Health, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China; dlaville@ips.ac.cn gxmeng@ips.ac.cn. CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China. Pasteurien College, Soochow University, Suzhou, Jiangsu 215006, China.

MeSH

Angiotensin-Converting Enzyme 2AnimalsCOVID-19HEK293 CellsHost-Pathogen InteractionsHumansMembrane FusionMiceProtein BindingProteolysisSARS-CoV-2Spike Glycoprotein, CoronavirusVirus Internalization

Pub Type(s)

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

Language

eng

PubMed ID

34930824

Citation

Yu, Shi, et al. "SARS-CoV-2 Spike Engagement of ACE2 Primes S2' Site Cleavage and Fusion Initiation." Proceedings of the National Academy of Sciences of the United States of America, vol. 119, no. 1, 2022.
Yu S, Zheng X, Zhou B, et al. SARS-CoV-2 spike engagement of ACE2 primes S2' site cleavage and fusion initiation. Proc Natl Acad Sci U S A. 2022;119(1).
Yu, S., Zheng, X., Zhou, B., Li, J., Chen, M., Deng, R., Wong, G., Lavillette, D., & Meng, G. (2022). SARS-CoV-2 spike engagement of ACE2 primes S2' site cleavage and fusion initiation. Proceedings of the National Academy of Sciences of the United States of America, 119(1). https://doi.org/10.1073/pnas.2111199119
Yu S, et al. SARS-CoV-2 Spike Engagement of ACE2 Primes S2' Site Cleavage and Fusion Initiation. Proc Natl Acad Sci U S A. 2022 01 4;119(1) PubMed PMID: 34930824.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - SARS-CoV-2 spike engagement of ACE2 primes S2' site cleavage and fusion initiation. AU - Yu,Shi, AU - Zheng,Xu, AU - Zhou,Bingjie, AU - Li,Juan, AU - Chen,Mengdan, AU - Deng,Rong, AU - Wong,Gary, AU - Lavillette,Dimitri, AU - Meng,Guangxun, PY - 2021/11/15/accepted PY - 2021/12/21/entrez PY - 2021/12/22/pubmed PY - 2022/1/5/medline KW - ACE2 KW - S2′ KW - SARS-CoV-2 spike KW - membrane fusion KW - spike protein JF - Proceedings of the National Academy of Sciences of the United States of America JO - Proc Natl Acad Sci U S A VL - 119 IS - 1 N2 - The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has resulted in tremendous loss worldwide. Although viral spike (S) protein binding of angiotensin-converting enzyme 2 (ACE2) has been established, the functional consequences of the initial receptor binding and the stepwise fusion process are not clear. By utilizing a cell-cell fusion system, in complement with a pseudoviral infection model, we found that the spike engagement of ACE2 primed the generation of S2' fragments in target cells, a key proteolytic event coupled with spike-mediated membrane fusion. Mutagenesis of an S2' cleavage site at the arginine (R) 815, but not an S2 cleavage site at arginine 685, was sufficient to prevent subsequent syncytia formation and infection in a variety of cell lines and primary cells isolated from human ACE2 knock-in mice. The requirement for S2' cleavage at the R815 site was also broadly shared by other SARS-CoV-2 spike variants, such as the Alpha, Beta, and Delta variants of concern. Thus, our study highlights an essential role for host receptor engagement and the key residue of spike for proteolytic activation, and uncovers a targetable mechanism for host cell infection by SARS-CoV-2. SN - 1091-6490 UR - https://www.unboundmedicine.com/medline/citation/34930824/SARS_CoV_2_spike_engagement_of_ACE2_primes_S2'_site_cleavage_and_fusion_initiation_ DB - PRIME DP - Unbound Medicine ER -