Overview of Nucleocapsid-Targeting Vaccines against COVID-19 DOI Creative Commons
Alexandra Rak,

Irina Isakova-Sivak,

Larisa Rudenko

et al.

Vaccines, Journal Year: 2023, Volume and Issue: 11(12), P. 1810 - 1810

Published: Dec. 3, 2023

The new SARS-CoV-2 coronavirus, which emerged in late 2019, is a highly variable causative agent of COVID-19, contagious respiratory disease with potentially severe complications. Vaccination considered the most effective measure to prevent spread and complications this infection. Spike (S) protein-based vaccines were very successful preventing COVID-19 caused by ancestral strain; however, their efficacy was significantly reduced when coronavirus variants antigenically different from original strain circulation. This due high variability major viral antigen escape immunity infection or vaccination spike-targeting vaccines. nucleocapsid protein (N) much more conserved than spike has therefore attracted attention scientists as promising target for broad-spectrum vaccine development. Here, we summarized current data on various N-based that have been tested animal challenge models clinical trials. Despite conservatism N protein, mutations gradually occurring sequence can affect its protective properties. During three years pandemic, at least 12 arisen sequence, affecting 40 known immunogenic T-cell epitopes, so antigenicity recent may be altered. fact should taken into account limitation development cross-reactive based N-protein.

Language: Английский

SARS-CoV-2 biology and host interactions DOI
Silvio Steiner, Annika Kratzel, G. Tuba Barut

et al.

Nature Reviews Microbiology, Journal Year: 2024, Volume and Issue: 22(4), P. 206 - 225

Published: Jan. 15, 2024

Language: Английский

Citations

85
Interaction between host G3BP and viral nucleocapsid protein regulates SARS-CoV-2 replication and pathogenicity DOI Creative Commons
Zemin Yang, Bryan A. Johnson, Victoria Meliopoulos

et al.

Cell Reports, Journal Year: 2024, Volume and Issue: 43(3), P. 113965 - 113965

Published: March 1, 2024

G3BP1/2 are paralogous proteins that promote stress granule formation in response to cellular stresses, including viral infection. The nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) inhibits assembly and interacts with via an ITFG motif, residue F17, the N protein. Prior studies examining impact G3PB1-N interaction on SARS-CoV-2 replication have produced inconsistent findings, role this pathogenesis is unknown. Here, we use structural biochemical analyses define residues required for G3BP1-N structure-guided mutagenesis selectively disrupt interaction. We find N-F17A mutation causes highly specific loss G3BP1/2. fails inhibit cells, has decreased replication, pathology vivo. Further mechanistic indicate N-F17-mediated promotes infection by limiting sequestration genomic RNA (gRNA) into granules.

Language: Английский

Citations

25
Proteomic analysis of SARS-CoV-2 particles unveils a key role of G3BP proteins in viral assembly DOI Creative Commons

Émilie Murigneux,

Laurent Softic,

C. Aubé

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Jan. 20, 2024

Language: Английский

Citations

17
A conserved oligomerization domain in the disordered linker of coronavirus nucleocapsid proteins DOI Creative Commons
Huaying Zhao, Di Wu, Sergio A. Hassan

et al.

Science Advances, Journal Year: 2023, Volume and Issue: 9(14)

Published: April 5, 2023

The nucleocapsid (N-)protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has a key role in viral assembly and scaffolding the RNA. It promotes liquid-liquid phase separation (LLPS), forming dense droplets that support ribonucleoprotein particles with as-of-yet unknown macromolecular architecture. Combining biophysical experiments, molecular dynamics simulations, analysis mutational landscape, we describe heretofore oligomerization site contributes to LLPS, is required for higher-order protein-nucleic acid complexes, coupled large-scale conformational changes N-protein upon nucleic binding. self-association interface located leucine-rich sequence intrinsically disordered linker between folded domains formed by transient helices assembling into trimeric coiled-coils. Critical residues stabilizing hydrophobic electrostatic interactions adjacent are highly protected against mutations viable SARS-CoV-2 genomes, motif conserved across related coronaviruses, thus presenting target antiviral therapeutics.

Language: Английский

Citations

32
Phase-separated nucleocapsid protein of SARS-CoV-2 suppresses cGAS-DNA recognition by disrupting cGAS-G3BP1 complex DOI Creative Commons
Sihui Cai,

Chenqiu Zhang,

Zhen Zhuang

et al.

Signal Transduction and Targeted Therapy, Journal Year: 2023, Volume and Issue: 8(1)

Published: April 26, 2023

Currently, the incidence and fatality rate of SARS-CoV-2 remain continually high worldwide. COVID-19 patients infected with exhibited decreased type I interferon (IFN-I) signal, along limited activation antiviral immune responses as well enhanced viral infectivity. Dramatic progresses have been made in revealing multiple strategies employed by impairing canonical RNA sensing pathways. However, it remains to be determined about antagonism cGAS-mediated IFN during infection. In current study, we figure out that infection leads accumulation released mitochondria DNA (mtDNA), which turn triggers cGAS activate IFN-I signaling. As countermeasures, nucleocapsid (N) protein restricts recognition capacity impair cGAS-induced Mechanically, N disrupts assembly its co-factor G3BP1 undergoing DNA-induced liquid-liquid phase separation (LLPS), subsequently impairs double-strand (dsDNA) detection ability cGAS. Taken together, our findings unravel a novel antagonistic strategy reduces DNA-triggered pathway through interfering cGAS-DNA separation.

Language: Английский

Citations

32
Assembly of SARS-CoV-2 nucleocapsid protein with nucleic acid DOI Creative Commons
Huaying Zhao, Abdullah M. Syed, Mir M. Khalid

et al.

Nucleic Acids Research, Journal Year: 2024, Volume and Issue: 52(11), P. 6647 - 6661

Published: April 8, 2024

Abstract The viral genome of SARS-CoV-2 is packaged by the nucleocapsid (N-)protein into ribonucleoprotein particles (RNPs), 38 ± 10 which are contained in each virion. Their architecture has remained unclear due to pleomorphism RNPs, high flexibility N-protein intrinsically disordered regions, and highly multivalent interactions between RNA binding sites both N-terminal (NTD) C-terminal domain (CTD). Here we explore critical interaction motifs RNPs applying a combination biophysical techniques ancestral mutant proteins different nucleic acids an vitro assay for RNP formation, examining protein variants assembly assay. We find that acid-bound dimers oligomerize via recently described protein–protein interface presented transient helix its long linker region NTD CTD. resulting hexameric complexes stabilized protein-nucleic acid establish crosslinks dimeric subunits. Assemblies CTD offering more than one site stem–loop RNA. Our study suggests model where scaffolding at density on followed cooperative multimerization through linker.

Language: Английский

Citations

11
Phosphorylation in the Ser/Arg-rich region of the nucleocapsid of SARS-CoV-2 regulates phase separation by inhibiting self-association of a distant helix DOI Creative Commons

Hannah Stuwe,

Patrick N. Reardon,

Zhen Yu

et al.

Journal of Biological Chemistry, Journal Year: 2024, Volume and Issue: 300(6), P. 107354 - 107354

Published: May 7, 2024

The nucleocapsid protein (N) of SARS-CoV-2 is essential for virus replication, genome packaging, evading host immunity, and maturation. N a multidomain composed an independently folded monomeric N-terminal domain that the primary site RNA binding, dimeric C-terminal efficient phase separation condensate formation with RNA. domains are separated by disordered Ser/Arg-rich region preceding self-associating Leu-rich helix. Phosphorylation in Ser/Arg infected cells decreases viscosity N:RNA condensates promoting viral replication immune evasion. molecular level effect phosphorylation, however, missing from our current understanding. Using NMR spectroscopy analytical ultracentrifugation we show phosphorylation destabilizes helix 30 amino acids distant site. gel shift assays demonstrate binding linker dampened whereas to full-length not significantly affected presumably due retained strong interactions domain. Introducing switchable replace confirms importance self-association droplet suggests only increases solubility positively charged elongated as observed other proteins but can also inhibit These data highlight both at local sites hydrophobic regulating liquid-liquid entire protein.

Language: Английский

Citations

11
Liquid-liquid phase separation of nucleocapsid proteins during SARS-CoV-2 and HIV-1 replication DOI Creative Commons

Bao-An Chau,

Venessa Chen,

Alan Cochrane

et al.

Cell Reports, Journal Year: 2022, Volume and Issue: 42(1), P. 111968 - 111968

Published: Dec. 26, 2022

The leap of retroviruses and coronaviruses from animal hosts to humans has led two ongoing pandemics tens millions deaths worldwide. Retrovirus coronavirus nucleocapsid proteins have been studied extensively as potential drug targets due their central roles in virus replication, among which is capacity bind respective genomic RNAs for packaging into nascent virions. This review focuses on fundamental studies these how intrinsic abilities condense through liquid-liquid phase separation (LLPS) contribute viral replication. Therapeutic targeting condensates methodological advances are also described address future questions contributes

Language: Английский

Citations

34
Modulation of biophysical properties of nucleocapsid protein in the mutant spectrum of SARS-CoV-2 DOI Creative Commons

Ai Nguyen,

Huaying Zhao,

Dulguun Myagmarsuren

et al.

eLife, Journal Year: 2024, Volume and Issue: 13

Published: Feb. 6, 2024

Genetic diversity is a hallmark of RNA viruses and the basis for their evolutionary success. Taking advantage uniquely large genomic database SARS-CoV-2, we examine impact mutations across spectrum viable amino acid sequences on biophysical phenotypes highly expressed multifunctional nucleocapsid protein. We find variation in physicochemical parameters its extended intrinsically disordered regions (IDRs) sufficient to allow local plasticity, but also observe functional constraints that similarly occur related coronaviruses. In experiments with several N-protein species carrying associated major variants, point IDRs can have nonlocal modulate thermodynamic stability, secondary structure, protein oligomeric state, particle formation, liquid-liquid phase separation. Omicron variant, distant different compensatory effects shifting delicate balance interactions controlling assembly properties, include creation new protein-protein interaction interface N-terminal IDR through defining P13L mutation. A picture emerges where genetic accompanied by significant characteristics species, particular IDRs.

Language: Английский

Citations

8
Detection of the SARS-CoV-2 Nucleoprotein by Electrochemical Biosensor based on Molecularly Imprinted Polypyrrole Formed on Self-Assembled Monolayer DOI
Viktorija Liustrovaitė, Vilma Ratautaitė, Almira Ramanavičienė

et al.

Biosensors and Bioelectronics, Journal Year: 2024, Volume and Issue: unknown, P. 117092 - 117092

Published: Dec. 1, 2024

Language: Английский

Citations

8