Functional benefit of structural disorder for the replication of measles, Nipah and Hendra viruses DOI
Frank Gondelaud, Giulia Pesce, Juliet F. Nilsson

et al.

Essays in Biochemistry, Journal Year: 2022, Volume and Issue: 66(7), P. 915 - 934

Published: Sept. 23, 2022

Abstract Measles, Nipah and Hendra viruses are severe human pathogens within the Paramyxoviridae family. Their non-segmented, single-stranded, negative-sense RNA genome is encapsidated by nucleoprotein (N) a helical nucleocapsid that substrate used viral RNA-dependent-RNA-polymerase (RpRd) for transcription replication. The RpRd complex made of large protein (L) phosphoprotein (P), latter serving as an obligate polymerase cofactor chaperon N. Both N P proteins enriched in intrinsically disordered regions (IDRs), i.e. devoid stable secondary tertiary structure. possesses C-terminal IDR (NTAIL), while consists large, N-terminal domain (NTD) (CTD) encompassing alternating ordered regions. V W proteins, two non-structural encoded gene via mechanism co-transcriptional edition mRNA, prevalently too, sharing with NTD. They key players evasion host antiviral response were shown to phase separate form amyloid-like fibrils vitro. In this review, we summarize available information on IDRs N, P, from these three model paramyxoviruses describe their molecular partnership. We discuss functional benefit disorder virus replication light critical role affording promiscuity, multifunctionality, fine regulation interaction strength, scaffolding functions promoting liquid–liquid separation fibrillation.

Language: Английский

Intrinsic factors behind long COVID: IV. Hypothetical roles of the SARS‐CoV‐2 nucleocapsid protein and its liquid–liquid phase separation DOI

Ahmed Eltayeb,

Faisal Al‐Sarraj, Mona G. Alharbi

et al.

Journal of Cellular Biochemistry, Journal Year: 2024, Volume and Issue: 125(3)

Published: Feb. 13, 2024

Abstract When the SARS‐CoV‐2 virus infects humans, it leads to a condition called COVID‐19 that has wide spectrum of clinical manifestations, from no symptoms acute respiratory distress syndrome. The initiates damage by attaching ACE‐2 protein on surface endothelial cells line blood vessels and using these as hosts for replication. Reactive oxygen species levels are increased during viral replication, which oxidative stress. About three‐fifths (~60%) people who get infected with eradicate their body after 28 days recover normal activity. However, large fraction (~40%) suffer various (anosmia and/or ageusia, fatigue, cough, myalgia, cognitive impairment, insomnia, dyspnea, tachycardia) beyond 12 weeks diagnosed syndrome long COVID. Long‐term studies in group contracted have been contrasted noninfected matched people. A subset can be distinguished set cytokine markers persistent, low‐grade inflammation often self‐report two or more bothersome symptoms. No medication alleviate efficiently. Coronavirus nucleocapsid proteins investigated extensively potential drug targets due key roles among is ability bind respective genomic RNAs incorporation into emerging virions. This review highlights basic its undergo liquid–liquid phase separation. We hypothesize this separation may contribute hypothesis unlocks new investigation angles could potentially open novel avenues better understanding COVID treating condition.

Language: Английский

Citations

6
Scaffolding viral protein NC nucleates phase separation of the HIV-1 biomolecular condensate DOI Creative Commons
Anne Monette,

Meijuan Niu,

Maya Nijhoff Asser

et al.

Cell Reports, Journal Year: 2022, Volume and Issue: 40(8), P. 111251 - 111251

Published: Aug. 1, 2022

Membraneless biomolecular condensates (BMCs) contribute to the replication of a growing number viruses but remain be functionally characterized. Previously, we demonstrated that pan-retroviral nucleocapsid (NC) proteins phase separated into regulating virus assembly. Here discover intrinsically disordered human immunodeficiency virus-type 1 (HIV-1) core condense with viral genomic RNA (vRNA) assemble as BMCs attaining geometry characteristic reverse transcription complexes. We explore predisposition, mechanisms, and pharmacologic sensitivity HIV-1 in living cells. vRNA-interacting NC were found scaffolds onto which client capsid, transcriptase, integrase assemble. exhibit fundamental characteristics are drug-sensitive. Lastly, protease-mediated maturation Gag Gag-Pol precursor yield abundant visible This study redefines components fluid advances our understanding nature cores during ingress.

Language: Английский

Citations

28
Protein conformation and biomolecular condensates DOI Creative Commons
Diego S. Vazquez,

Pamela L. Toledo,

Alejo R. Gianotti

et al.

Current Research in Structural Biology, Journal Year: 2022, Volume and Issue: 4, P. 285 - 307

Published: Jan. 1, 2022

Protein conformation and cell compartmentalization are fundamental concepts subjects of vast scientific endeavors. In the last two decades, we have witnessed exciting advances that unveiled conjunction these concepts. An avalanche studies highlighted central role biomolecular condensates in membraneless subcellular permits spatiotemporal organization regulation myriads simultaneous biochemical reactions macromolecular interactions. These also shown condensation, driven by multivalent intermolecular interactions, is mediated order-disorder transitions protein domain architecture. Conceptually, condensation a distinct level conformational landscape which collective folding large collections molecules takes place. Biomolecular arise physical process phase separation comprise variety bodies ranging from organelles to liquid solid-like conglomerates, spanning lengths mesoscopic clusters (nanometers) micrometer-sized objects. this review, summarize discuss recent work on assembly, composition, conformation, material properties, thermodynamics, regulation, functions bodies. We review conceptual framework for future dynamics condensed proteins cellular processes.

Language: Английский

Citations

24
Flexibility of the Rotavirus NSP2 C-Terminal Region Supports Factory Formation via Liquid-Liquid Phase Separation DOI

Sarah L. Nichols,

Emil M. Nilsson,

Heather M. Brown

et al.

Journal of Virology, Journal Year: 2023, Volume and Issue: 97(2)

Published: Feb. 7, 2023

Viruses often condense the materials needed for their replication into discrete intracellular factories. For rotaviruses, agents of severe gastroenteritis in children, factory formation is mediated part by an octameric protein called NSP2.

Language: Английский

Citations

14
Subversion of selective autophagy for the biogenesis of tombusvirus replication organelles inhibits autophagy DOI Creative Commons
Yuanrong Kang, Wenwu Lin, Peter D. Nagy

et al.

PLoS Pathogens, Journal Year: 2024, Volume and Issue: 20(3), P. e1012085 - e1012085

Published: March 14, 2024

Elaborate viral replication organelles (VROs) are formed to support positive-strand RNA virus in infected cells. VRO formation requires subversion of intracellular membranes by proteins. Here, we showed that the key ATG8f autophagy protein and NBR1 selective receptor were co-opted Tomato bushy stunt (TBSV) closely-related carnation Italian ringspot virus. Knockdown or plants led reduced tombusvirus replication, suggesting pro-viral function for autophagy. BiFC proximity-labeling experiments TBSV p33 interacted with recruit them VROs. In addition, observed several core proteins, such as ATG1a, ATG4, ATG5, ATG101 plant-specific SH3P2 adaptor proteins also re-localized VROs, hijacks machinery plant We demonstrated components facilitated recruitment VPS34 PI3 kinase enrichment phospholipids, phosphatidylethanolamine PI3P phosphoinositide membranes. Hijacking into VROs inhibition autophagic flux. found a fraction subverted was sequestered biomolecular condensates associated propose VRO-associated trap those taking away from pathway. Overall, tombusviruses hijack provide phospholipid-rich regulate antiviral

Language: Английский

Citations

4
Exploring the expanding universe of host-virus interactions mediated by viral RNA DOI Creative Commons
Alfredo Castelló, Lucı́a Álvarez, Wael Kamel

et al.

Molecular Cell, Journal Year: 2024, Volume and Issue: 84(19), P. 3706 - 3721

Published: Oct. 1, 2024

Language: Английский

Citations

4
Genetically Recoding Respiratory Syncytial Virus to Visualize Nucleoprotein Dynamics and Virion Assembly DOI Creative Commons
Margaret D. Mitrovich, Michael D. Vahey

ACS Infectious Diseases, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

RNA viruses possess small genomes encoding a limited repertoire of essential and often multifunctional proteins. Although genetically tagging viral proteins provides powerful tool for dissecting mechanisms replication infection, it remains challenge. Here, we leverage genetic code expansion to develop recoded strain respiratory syncytial virus (RSV) in which the nucleoprotein is site-specifically modified with noncanonical amino acid. The resulting replicates exclusively cells capable amber stop codon suppression amenable labeling tetrazine-modified fluorophores, achieving high signal background. Virus labeled functional, retaining ∼70% infectivity relative unlabeled controls. We this visualize RSV assembly, capturing transfer complexes from cytoplasmic condensates directly budding filaments at cell surface compartments containing Collectively, these results suggest multiple pathways assembly establish framework that may be extended other nucleoproteins.

Language: Английский

Citations

0
Fluorescence Loss After Photoactivation (FLAPh): A Pulse-Chase Cellular Assay for Understanding Kinetics and Dynamics of Viral Inclusions DOI
Temitope Akhigbe Etibor, Tiago Paixão, Maria João Amorim

et al.

Methods in molecular biology, Journal Year: 2025, Volume and Issue: unknown, P. 125 - 140

Published: Jan. 1, 2025

Language: Английский

Citations

0
HBV polymerase recruits the phosphatase PP1 to dephosphorylate HBc-Ser170 to complete encapsidation DOI Creative Commons

Chi-Ling Hsieh,

Lung‐Ji Chang, Pei‐Jer Chen

et al.

PLoS Pathogens, Journal Year: 2025, Volume and Issue: 21(2), P. e1012905 - e1012905

Published: Feb. 11, 2025

The HBV core (HBc) protein contains an N-terminal domain (NTD) for capsid assembly and arginine-rich C-terminal (CTD) pregenomic RNA (pgRNA) encapsidation. Phosphorylation of the HBc CTD, especially at Ser162 Ser170, is essential nucleation with polymerase (Pol) to initiate pgRNA As capsids mature, CTD undergoes dephosphorylation, suggesting involvement a phosphatase in late stage encapsidation, which remains be determined. Using C-S170 antibody specific non-phosphorylated HBc-Ser170, we observed transition from phosphorylated dephosphorylated state during packaging. Pol-dependent dephosphorylation HBc-Ser170 was confirmed by substitution one single amino acid Val782 RNase H domain, abolished HBc-Ser170. Immunoprecipitation, mass spectrometry analyses, structural analyses showed that recruitment host PP1 dependent on Pol-Val782 domain. This does not require but Pol via epsilon signal, Pol-pgRNA complex plays key role recruitment. Pol-pgRNA-PP1-mediated completion encapsidation appears associated endosomes/multivesicular bodies (MVBs). Therefore, may play dual initially bringing recruiting later packaging into capsids. These findings only decipher mechanism Pol-mediated regulates encapsulation, also reveal possibility as potential target antiviral development.

Language: Английский

Citations

0
Extending fluorescence anisotropy to large complexes using reversibly switchable proteins DOI Creative Commons
Andrea Volpato, Dirk Ollech, Jonatan Alvelid

et al.

Nature Biotechnology, Journal Year: 2022, Volume and Issue: 41(4), P. 552 - 559

Published: Oct. 10, 2022

Abstract The formation of macromolecular complexes can be measured by detection changes in rotational mobility using time-resolved fluorescence anisotropy. However, this method is limited to relatively small molecules (~0.1–30 kDa), excluding the majority human proteome and its complexes. We describe selective anisotropy with reversibly switchable states (STARSS), which overcomes limitation extends observable mass range more than three orders magnitude. STARSS based on long-lived reversible molecular transitions fluorescent proteins resolve slow diffusivity large used probe several cells, including chromatin, retroviral Gag lattice activity-regulated cytoskeleton-associated protein oligomers. Because arbitrarily structures, it generally applicable entire proteome.

Language: Английский

Citations

17