The sequence at Spike S1/S2 site enables cleavage by furin and phospho-regulation in SARS-CoV2 but not in SARS-CoV1 or MERS-CoV

Örd, Mihkel; Faustova, Ilona; Loog, Mart

doi:10.1038/s41598-020-74101-0

Cited by 149 publications

(132 citation statements)

References 52 publications

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“…In this study, we present a detailed characterization of the cleavage patterns, protein stability, and cell-cell fusion function of the SARS-CoV-2 S protein, as well as analysis of mutations within the S2 subunit that may affect these important protein properties. Consistent with recently published work [ 26 , 35 , 47 , 50 , 51 , 66 ], our analysis confirms that S is readily cleaved at the S1/S2 border in a variety of mammalian cell lines. Additionally, we show for the first time, that cleavage occurs in a bat cell line similar to the SARS-CoV-2 reservoir species.…”

Section: Discussionsupporting

confidence: 92%

Effect of mutations in the SARS-CoV-2 spike protein on protein stability, cleavage, and cell-cell fusion function

Barrett

Neal

Edmonds

et al. 2021

Preprint

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The SARS-CoV-2 spike protein (S) is the sole viral protein responsible for both viral binding to a host cell and the membrane fusion event needed for cell entry. In addition to facilitating fusion needed for viral entry, S can also drive cell-cell fusion, a pathogenic effect observed in the lungs of SARS-CoV-2 infected patients. While several studies have investigated S requirements involved in viral particle entry, examination of S stability and factors involved in S cell-cell fusion remain limited. We demonstrate that S must be processed at the S1/S2 border in order to mediate cell-cell fusion, and that mutations at potential cleavage sites within the S2 subunit alter S processing at the S1/S2 border, thus preventing cell-cell fusion. We also identify residues within the internal fusion peptide and the cytoplasmic tail that modulate S cell-cell fusion. Additionally, we examine S stability and protein cleavage kinetics in a variety of mammalian cell lines, including a bat cell line related to the likely reservoir species for SARS-CoV-2, and provide evidence that proteolytic processing alters the stability of the S trimer. This work therefore offers insight into S stability, proteolytic processing, and factors that mediate S cell-cell fusion, all of which help give a more comprehensive understanding of this highly sought-after therapeutic target.

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Section: Discussionsupporting

confidence: 92%

Effect of mutations in the SARS-CoV-2 spike protein on protein stability, cleavage, and cell-cell fusion function

Barrett

Neal

Edmonds

et al. 2021

Preprint

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“…The S1-S2 site is where the protease, furin, cleaves. The S1 unit binds to the ACE2 receptor [ 59 ] and the S2 unit mediates fusion of the viral and cellular membranes [ 60 ]. A more detailed discussion is presented in the next section.…”

Section: Discussionmentioning

confidence: 99%

“…These sequential steps of the trimer include: (i) closed conformation, (ii) open conformation (only one receptor binding domain (RBD) points “up” the other two RBD remain “closed”) but unbound to ACE2 (similar to the MERS CoV spike structure provided with PDB ID: 5X5U), (iii) one ACE2 bound to the RBD of one protomer, (iv) two ACE2 bound to two protomers at the RBDs, (v) three ACE2 bound to three protomers at the RBDs, and (vi) release of the monomeric S1-ACE2 complex (S1 unit includes residues 14–685). The S1 unit is first cleaved by the protease, furin [ 59 , 64 ], from the host cell [ 63 , 71 ]. The serine protease, transmembrane protease serine 2 (TMPRSS2), is also known to prime the spike protein for cell entry by cleaving at the S2′ site (position 816) [ 61 ].…”

Section: Discussionmentioning

confidence: 99%

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A Biochemical Perspective of the Nonstructural Proteins (NSPs) and the Spike Protein of SARS CoV-2

Yoshimoto

2021

Protein J

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The global pandemic that shut down the world in 2020 was caused by the virus, SARS CoV-2. The chemistry of the various nonstructural proteins (NSP3, NSP5, NSP12, NSP13, NSP14, NSP15, NSP16) of SARS CoV-2 is discussed. Secondly, a recent major focus of this pandemic is the variant strains of SARS CoV-2 that are increasingly occurring and more transmissible. One strain, called “D614G”, possesses a glycine (G) instead of an aspartate (D) at position 614 of the spike protein. Additionally, other emerging strains called “501Y.V1” and “501Y.V2” have several differences in the receptor binding domain of the spike protein (N501Y) as well as other locations. These structural changes may enhance the interaction between the spike protein and the ACE2 receptor of the host, increasing infectivity. The global pandemic caused by SARS CoV-2 is a rapidly evolving situation, emphasizing the importance of continuing the efforts to interrogate and understand this virus. Supplementary Information The online version contains supplementary material available at 10.1007/s10930-021-09967-8.

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“…134 A particular relevance of this site stems from the fact that sequence of the S1/S2 site enables cleavage by furin in SARS-CoV-2 but not in SARS-CoV or MERS-CoV viruses. 135 The experimental data also showed that SARS-CoV S-mediated virus entry is based on sequential proteolytic cleavage at two distinct sites, with cleavage at the S1/S2 boundary (R667) promoting subsequent cleavage at the S2′ position (R797) triggering membrane fusion. 136 Interestingly, our results indicated a high coevolutionary signal for R667 at the S1/S2 boundary but only a moderate Cscore for the conserved R797 position ( Figure 5).…”

Section: Coevolutionary Analysis Of the Sars-cov-2 Proteins Reveals Rmentioning

confidence: 94%

Coevolutionary Analysis and Perturbation-Based Network Modeling of the SARS-CoV-2 Spike Protein Complexes with Antibodies: Binding-Induced Control of Dynamics, Allosteric Interactions and Signaling

Verkhivker

Paola

2021

Preprint

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The structural and biochemical studies of the SARS-CoV-2 spike glycoproteins and complexes with highly potent antibodies have revealed multiple conformation-dependent epitopes highlighting the link between conformational plasticity of spike proteins and capacity for eliciting specific binding and broad neutralization responses. In this study, we used coevolutionary analysis, molecular simulations, and perturbation-based hierarchical network modeling of the SARS-CoV-2 S complexes with H014, S309, S2M11 and S2E12 antibodies targeting distinct epitopes to explore molecular mechanisms underlying binding-induced modulation of dynamics, stability and allosteric signaling in the spike protein trimers. The results of this study revealed key regulatory centers that can govern allosteric interactions and communications in the SARS-CoV-2 spike proteins. Through coevolutionary analysis of the SARS-CoV-2 spike proteins, we identified highly coevolving hotspots and functional clusters forming coevolutionary networks. The results revealed significant coevolutionary couplings between functional regions separated by the medium-range distances which may help to facilitate a functional cross-talk between distant allosteric regions in the SARS-CoV-2 spike complexes with antibodies. We also discovered a potential mechanism by which antibody-specific targeting of coevolutionary centers can allow for efficient modulation of allosteric interactions and signal propagation between remote functional regions. Using a hierarchical network modeling and perturbation-response scanning analysis, we demonstrated that binding of antibodies could leverage direct contacts with coevolutionary hotspots to allosterically restore and enhance couplings between spatially separated functional regions, thereby protecting the spike apparatus from membrane fusion. The results of this study also suggested that antibody binding can induce a switch from a moderately cooperative population-shift mechanism, governing structural changes of the ligand-free SARS-CoV-2 spike protein, to antibody-induced highly cooperative mechanism that can better withstand mutations in the functional regions without significant deleterious consequences for protein function. This study provides a novel insight into allosteric regulatory mechanisms of SARS-CoV-2 S proteins, showing that antibodies can modulate allosteric interactions and signaling of spike proteins, providing a plausible strategy for therapeutic intervention by targeting specific hotspots of allosteric interactions in the SARS-CoV-2 proteins.

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The sequence at Spike S1/S2 site enables cleavage by furin and phospho-regulation in SARS-CoV2 but not in SARS-CoV1 or MERS-CoV

Cited by 149 publications

References 52 publications

Effect of mutations in the SARS-CoV-2 spike protein on protein stability, cleavage, and cell-cell fusion function

Effect of mutations in the SARS-CoV-2 spike protein on protein stability, cleavage, and cell-cell fusion function

A Biochemical Perspective of the Nonstructural Proteins (NSPs) and the Spike Protein of SARS CoV-2

Coevolutionary Analysis and Perturbation-Based Network Modeling of the SARS-CoV-2 Spike Protein Complexes with Antibodies: Binding-Induced Control of Dynamics, Allosteric Interactions and Signaling

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