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

Nichols, Sarah L; Nilsson, Emil M.; Brown-Harding, Heather; LaConte, Leslie E. W.; Acker, Julia; Borodavka, Alexander; Esstman, Sarah McDonald

doi:10.1128/jvi.00039-23

Cited by 12 publications

(17 citation statements)

References 54 publications

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“…Notably, the phosphorylation of NSP2 has also been implicated in viroplasm formation, as evidenced by the delayed formation of viroplasms observed in a rRV harboring an NSP2 S313D phosphomimetic mutant [75]. Additionally, studies using a mutant NSP2 harboring a lysine-to-glutamic acid change in the C-terminal region revealed the importance of a flexible tail in viroplasm biogenesis and coalescence properties [76].…”

Section: Nsp2mentioning

confidence: 99%

Host Requirements for Formation and Dynamics of Rotavirus Viroplasms

Vetter,

Lee,

Eichwald

2024

Preprint

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Rotavirus (RV) replicates within viroplasms, membrane-less electron-dense globular cytosolic inclusions with liquid-liquid phase properties. In these structures occur the virus transcription, replication, and packaging of the virus genome in newly assembled double-layered particles. The viroplasms are composed of virus proteins (NSP2, NSP5, NSP4, VP1, VP2, VP3, and VP6), single and double-stranded virus RNAs, and host components such as microtubules, perilipin-1 and chaperonins. The formation, coalescence, maintenance, and perinuclear localization of viroplasms rely on their association with the cytoskeleton. A stabilized microtubule network involving microtubules and kinesin Eg5 and dynein molecular motors is associated with NSP5, NSP2, and VP2, facilitating dynamic processes such as viroplasm coalescence and perinuclear localization. Key post-translation modifications, particularly phosphorylation events of RV proteins NSP5 and NSP2, play pivotal roles in orchestrating these interactions. Actin filaments also contribute, triggering the formation of the viroplasms through the association of soluble cytosolic VP4 with actin and the molecular motor myosin. This review explores the evolving understanding of RV replication, emphasizing the host requirements essential for viroplasm formation and highlighting their dynamic interplay within the host cell.

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Section: Nsp2mentioning

confidence: 99%

Host Requirements for Formation and Dynamics of Rotavirus Viroplasms

Vetter,

Lee,

Eichwald

2024

Preprint

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“…Notably, the phosphorylation of NSP2 has been implicated in viroplasm formation, as evidenced by the delayed formation of viroplasms observed in a rRV harboring an NSP2 S313D phosphomimetic mutant [81]. Additionally, studies using a mutant NSP2 harboring a lysine-to-glutamic acid change in the C-terminal region revealed the importance of a flexible tail in viroplasm biogenesis and coalescence properties [82].…”

Section: Nsp2mentioning

confidence: 99%

The Role of the Host Cytoskeleton in the Formation and Dynamics of Rotavirus Viroplasms

Vetter,

Lee,

Eichwald

2024

Viruses

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Rotavirus (RV) replicates within viroplasms, membraneless electron-dense globular cytosolic inclusions with liquid–liquid phase properties. In these structures occur the virus transcription, replication, and packaging of the virus genome in newly assembled double-layered particles. The viroplasms are composed of virus proteins (NSP2, NSP5, NSP4, VP1, VP2, VP3, and VP6), single- and double-stranded virus RNAs, and host components such as microtubules, perilipin-1, and chaperonins. The formation, coalescence, maintenance, and perinuclear localization of viroplasms rely on their association with the cytoskeleton. A stabilized microtubule network involving microtubules and kinesin Eg5 and dynein molecular motors is associated with NSP5, NSP2, and VP2, facilitating dynamic processes such as viroplasm coalescence and perinuclear localization. Key post-translation modifications, particularly phosphorylation events of RV proteins NSP5 and NSP2, play pivotal roles in orchestrating these interactions. Actin filaments also contribute, triggering the formation of the viroplasms through the association of soluble cytosolic VP4 with actin and the molecular motor myosin. This review explores the evolving understanding of RV replication, emphasizing the host requirements essential for viroplasm formation and highlighting their dynamic interplay within the host cell.

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“…Viroplasm-like inclusions are formed following coexpression of the negatively charged nonstructural phosphoprotein NSP5 (scaffold) and the positively charged RNA chaperone NSP2, the inner capsid viral protein VP2, or both NSP2 and VP2 (73)(74)(75)(76)(77). NSP5:NSP2 droplets also form in an in vitro reconstitution system at physiologically relevant low µM concentrations matching those achieved during infection (27,78). Previous views of viroplasm formation have been dominated by the idea that viral proteins (VPs) are recruited into these structures in a specific order (73,(75)(76)(77)(78)(79), yielding a particular multilayered organization (80).…”

Section: Early Replication Rotavirus Viral Factories Are Ribonucleopr...mentioning

confidence: 99%

“…NSP5:NSP2 droplets also form in an in vitro reconstitution system at physiologically relevant low µM concentrations matching those achieved during infection (27,78). Previous views of viroplasm formation have been dominated by the idea that viral proteins (VPs) are recruited into these structures in a specific order (73,(75)(76)(77)(78)(79), yielding a particular multilayered organization (80). While the exact organization of viroplasms is likely to be more complex than the concentric layers identified by super-resolution microscopy (80), multilayered organization is commonly seen in RNP condensates (18,81).…”

Section: Early Replication Rotavirus Viral Factories Are Ribonucleopr...mentioning

confidence: 99%

Seeing Biomolecular Condensates Through the Lens of Viruses

Borodavka

Acker

2023

Annual Review of Virology

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Phase separation of viral biopolymers is a key factor in the formation of cytoplasmic viral inclusions, known as sites of virus replication and assembly. This review describes the mechanisms and factors that affect phase separation in viral replication and identifies potential areas for future research. Drawing inspiration from studies on ribosome biogenesis, we compare the hierarchical coassembly of ribosomal RNAs and proteins in the nucleolus to the coordinated coassembly of viral RNAs and proteins taking place within viral factories formed by RNA viruses containing segmented genomes. We highlight the evidence supporting the role of biomolecular condensates in viral replication and how this new understanding is reshaping our views of virus assembly mechanisms. Future research of biomolecular condensates has the potential to uncover unexplored antiviral strategies targeting these phase-separated states. Expected final online publication date for the Annual Review of Virology, Volume 10 is September 2023. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

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

Abstract: 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 in part by an octameric protein called NSP2.

Cited by 12 publications

References 54 publications

Host Requirements for Formation and Dynamics of Rotavirus Viroplasms

Host Requirements for Formation and Dynamics of Rotavirus Viroplasms

The Role of the Host Cytoskeleton in the Formation and Dynamics of Rotavirus Viroplasms

Seeing Biomolecular Condensates Through the Lens of Viruses

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