Characterization of SARS-CoV-2 nucleocapsid protein reveals multiple functional consequences of the C-terminal domain

Wu, Chao; Qavi, Abraham J.; Hachim, Asamaa; Kavian, Niloufar; Cole, Aidan R; Moyle, Austin B.; Wagner, Nicole D.; Sweeney-Gibbons, Joyce; Rohrs, Henry W.; Gross, Michael L.; Peiris, J S Malik; Basler, Christopher F.; Farnsworth, Christopher W; Valkenburg, Sophie A.; Amarasinghe, Gaya K.; Leung, Daisy W.

doi:10.1016/j.isci.2021.102681

Cited by 77 publications

(94 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…N-CTD is responsible for type I interferon antagonism of the N protein. 21 CoronaVac appeared to elicit greater CD4 + and CD8 + T cell responses against the SARS-CoV2 structural peptide pool than BNT162b2 in a small group of adults, 31 and we also observed N-and M-specific T cell responses in adolescents receiving CC.…”

Section: Discussionsupporting

confidence: 52%

“…18 Additionally, non-S SARS-CoV-2 structural proteins, such as the nucleocapsid (N) and membrane (M), are associated with antibody responses in convalescent patients, and in fact, the C-terminal domain of N (N-CTD) is more specific to SARS-CoV-2. [19][20][21] Therefore, studies on immunogenicity outcomes that include these components are necessary.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Immunogenicity and reactogenicity of SARS-CoV-2 mRNA and inactivated vaccines in healthy adolescents

Lau

Duque

Wang

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

For SARS-CoV-2 vaccines, efficacy data for BNT162b2 but not CoronaVac are available in adolescents. Phase II/III studies focused on neutralizing antibody responses in adolescents, neglecting binding antibody and cellular responses that are also important against SARS-CoV-2. Therefore, we conducted a registered clinical study (NCT04800133) to establish immunobridging with various antibody and cellular immunity markers and to compare the immunogenicity and reactogenicity of these 2 vaccines in healthy adolescents. One-dose BNT162b2 outcomes were also assessed since it had been recommended in some localities due to the risk of myocarditis. Antibodies and T cell immune responses were non-inferior or similar in adolescents receiving 2 doses of BNT162b2 (BB, N=116) and CoronaVac (CC, N=123) versus adults after 2 doses of the same vaccine (BB, N=147; CC, N=141) but not in adolescents after 1 dose of BNT162b2 (B, N=116). CC induced SARS-CoV-2 nucleocapsid (N) and N C-terminal domain seroconversion in more adolescents than adults. Adverse reactions were mostly mild for both vaccines and more frequent for BNT162b2. We confirmed higher S, neutralizing, avidity and Fc receptor-binding antibody responses in adolescents receiving BB than CC. This is the first study to show similar induction of strong S-specific T cells by the 2 vaccines, in addition to N- and M-specific T cells induced by CoronaVac but not BNT162b2 in adolescents. The implications of the differential ability to induce S- and non-S-specific antibody and T cell responses on the durability of protection and protection against virus variants by BNT162b2 and CoronaVac, the 2 most used SARS-CoV-2 vaccines in the world, should be further investigated. Our results support the use of both vaccines in adolescents.

show abstract

Section: Discussionsupporting

confidence: 52%

Section: Introductionmentioning

confidence: 99%

Immunogenicity and reactogenicity of SARS-CoV-2 mRNA and inactivated vaccines in healthy adolescents

Lau

Duque

Wang

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…This sample showed a molecular mass of 89,5 ± 6.3 kDa and hydrodynamic radius of 8.0 ± 1.3 nm determined by SEC-MALS and DLS, respectively, consistent with a dimer in solution (Figure 1, C and D). Noteworthy, the hydrodynamic radius is close to the one recently reported for N protein purified under similar conditions [35]. Also, these results are in line with literature data showing that N protein readily oligomerizes into dimers [36,37].…”

Section: Dimers Of Full-length N Adopts An Extended Conformation In the Absence Of Rnasupporting

confidence: 91%

Structural dynamics of SARS-CoV-2 nucleocapsid protein induced by RNA binding

Ribeiro-Filho

Jara

Batista

et al. 2021

Preprint

View full text Add to dashboard Cite

The nucleocapsid (N) protein of the SARS-CoV-2 virus, the causal agent of COVID-19, is a multifunction phosphoprotein that plays critical roles in the virus life cycle, including transcription and packaging of the viral RNA. To play such diverse roles, the N protein has two structured RNA-binding modules, the N- (NTD) and C-terminal (CTD) domains, which are connected by an intrinsically disordered region. Despite the wealth of structural data available for the isolated NTD and CTD, how these domains are arranged in the full-length protein and how the oligomerization of N influences its RNA-binding activity remains largely unclear. Herein, using experimental data from electron microscopy and biochemical/biophysical techniques combined with molecular modeling and molecular dynamics simulations, we show that, in the absence of RNA, the N protein forms structurally dynamic dimers with the NTD and CTD arranged in extended conformations. In the presence of RNA, however, the N protein assumes a more compact conformation where the NTD and CTD are packed together. We also provide an octameric model for the full-length N bound to RNA that is consistent with electron microscopy images of the N protein in the presence of RNA. Together, our results shed new light on the dynamics and higher-order oligomeric structure of this versatile protein.

show abstract

“…The phase separation propensity of N protein has been subjected to extensive studies. N protein undergoes LLPS with RNA where the N-terminal RNA-binding domain, the central IDR, and C-terminal dimerization domain play an essential role [39,[189][190][191][192][193][194]. LLPS of N protein is modulated by phosphorylation at the serine/arginine-rich region by CDK1, GSK-3β, or SRPK1 [38,193,195].…”

Section: Sars-cov-2 Nucleocapsid Proteinmentioning

confidence: 99%

14-3-3 Proteins are Potential Regulators of Liquid–Liquid Phase Separation

Huang

Zheng

et al. 2022

Cell Biochem Biophys

View full text Add to dashboard Cite

The 14-3-3 family proteins are vital scaffold proteins that ubiquitously expressed in various tissues. They interact with numerous protein targets and mediate many cellular signaling pathways. The 14-3-3 binding motifs are often embedded in intrinsically disordered regions which are closely associated with liquid-liquid phase separation (LLPS). In the past ten years, LLPS has been observed for a variety of proteins and biological processes, indicating that LLPS plays a fundamental role in the formation of membraneless organelles and cellular condensates. While extensive investigations have been performed on 14-3-3 proteins, its involvement in LLPS is overlooked. To date, 14-3-3 proteins have not been reported to undergo LLPS alone or regulate LLPS of their binding partners. To reveal the potential involvement of 14-3-3 proteins in LLPS, in this review, we summarized the LLPS propensity of 14-3-3 binding partners and found that about one half of them may undergo LLPS spontaneously. We further analyzed the phase separation behavior of representative 14-3-3 binders and discussed how 14-3-3 proteins may be involved. By modulating the conformation and valence of interactions and recruiting other molecules, we speculate that 14-3-3 proteins can efficiently regulate the functions of their targets in the context of LLPS. Considering the critical roles of 14-3-3 proteins, there is an urgent need for investigating the involvement of 14-3-3 proteins in the phase separation process of their targets and the underling mechanisms.

show abstract

Characterization of SARS-CoV-2 nucleocapsid protein reveals multiple functional consequences of the C-terminal domain

Cited by 77 publications

References 55 publications

Immunogenicity and reactogenicity of SARS-CoV-2 mRNA and inactivated vaccines in healthy adolescents

Immunogenicity and reactogenicity of SARS-CoV-2 mRNA and inactivated vaccines in healthy adolescents

Structural dynamics of SARS-CoV-2 nucleocapsid protein induced by RNA binding

14-3-3 Proteins are Potential Regulators of Liquid–Liquid Phase Separation

Contact Info

Product

Resources

About