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dc.contributor.authorPapa, Guido
dc.contributor.authorPotgieter, Christiaan
dc.contributor.authorVenditti, Luca
dc.contributor.authorArnoldi, Francesca
dc.contributor.authorSchraner, Elisabeth M.
dc.date.accessioned2020-03-03T08:09:30Z
dc.date.available2020-03-03T08:09:30Z
dc.date.issued2020
dc.identifier.citationPapa, G. et al. 2020. Recombinant rotaviruses rescued by reverse genetics reveal the role of NSP5 hyperphosphorylation in the assembly of viral factories. Journal of virology, 94(1):#e01110-19. [https://doi .org/10.1128/JVI.01110-19]en_US
dc.identifier.issn0022-538X
dc.identifier.issn1098-5514 (Online)
dc.identifier.urihttp://hdl.handle.net/10394/34238
dc.identifier.urihttps://jvi.asm.org/content/94/1/e01110-19
dc.identifier.urihttps://doi.org/10.1128/JVI.01110-19
dc.description.abstractRotavirus (RV) replicates in round-shaped cytoplasmic viral factories, although how they assemble remains unknown. During RV infection, NSP5 undergoes hyperphosphorylation, which is primed by the phosphorylation of a single serine residue. The role of this posttranslational modification in the formation of viroplasms and its impact on virus replication remain obscure. Here, we investigated the role of NSP5 during RV infection by taking advantage of a modified fully tractable reverse-genetics system. A trans-complementing cell line stably producing NSP5 was used to generate and characterize several recombinant rotaviruses (rRVs) with mutations in NSP5. We demonstrate that an rRV lacking NSP5 was completely unable to assemble viroplasms and to replicate, confirming its pivotal role in rotavirus replication. A number of mutants with impaired NSP5 phosphorylation were generated to further interrogate the function of this posttranslational modification in the assembly of replication-competent viroplasms. We showed that the rRV mutant strains exhibited impaired viral replication and the ability to assemble round-shaped viroplasms in MA104 cells. Furthermore, we investigated the mechanism of NSP5 hyperphosphorylation during RV infection using NSP5 phosphorylation-negative rRV strains, as well as MA104-derived stable transfectant cell lines expressing either wild-type NSP5 or selected NSP5 deletion mutants. Our results indicate that NSP5 hyperphosphorylation is a crucial step for the assembly of round-shaped viroplasms, highlighting the key role of the C-terminal tail of NSP5 in the formation of replication-competent viral factories. Such a complex NSP5 phosphorylation cascade may serve as a paradigm for the assembly of functional viral factories in other RNA virusesen_US
dc.language.isoenen_US
dc.publisherASMen_US
dc.subjectNSP5en_US
dc.subjectReverse geneticsen_US
dc.subjectRotavirusen_US
dc.subjectViral factoriesen_US
dc.subjectViroplasmsen_US
dc.subjectProteinen_US
dc.subjectPhosphorylationen_US
dc.subjectRecombinant virusesen_US
dc.titleRecombinant rotaviruses rescued by reverse genetics reveal the role of NSP5 hyperphosphorylation in the assembly of viral factoriesen_US
dc.typeArticleen_US
dc.contributor.researchID10085637 - Potgieter, Abraham Christiaan


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