Rotavirus Particle Disassembly and Assembly In Vivo and In Vitro

Asensio-Cob, Dunia; Rodríguez, Javier M.; Luque, Daniel

doi:10.3390/v15081750

Cited by 10 publications

(4 citation statements)

References 184 publications

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“…The size distribution of viral particles has been earlier estimated for two other bunyaviruses, LACV ( 9 ) and UUKV ( 10 ). The amplitude of the size variation is comparable in all three bunyaviruses (including TSWV studied here) and higher than that of other segmented viruses which were shown to possess specific packaging mechanisms of their genome segments, such as, for example, influenza virus A ( 29 ) and rotaviruses ( 30 ). Notably, the size variation estimated here for TSWV is even more important than that of birnaviruses ( 31 ) which have been shown to encapsidate their two genomic segments in a nonspecific manner, often packaging more than one copy of each ( 32 ).…”

Section: Discussionmentioning

confidence: 72%

The genome of a bunyavirus cannot be defined at the level of the viral particle but only at the scale of the viral population

Yvon,

German,

Ullman

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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Bunyaviruses are enveloped negative or ambisense single-stranded RNA viruses with a genome divided into several segments. The canonical view depicts each viral particle packaging one copy of each genomic segment in one polarity named the viral strand. Several opposing observations revealed nonequal ratios of the segments, uneven number of segments per virion, and even packaging of viral complementary strands. Unfortunately, these observations result from studies often addressing other questions, on distinct viral species, and not using accurate quantitative methods. Hence, what RNA segments and strands are packaged as the genome of any bunyavirus remains largely ambiguous. We addressed this issue by first investigating the virion size distribution and RNA content in populations of the tomato spotted wilt virus (TSWV) using microscopy and tomography. These revealed heterogeneity in viral particle volume and amount of RNA content, with a surprising lack of correlation between the two. Then, the ratios of all genomic segments and strands were established using RNA sequencing and qRT-PCR. Within virions, both plus and minus strands (but no mRNA) are packaged for each of the three L, M, and S segments, in reproducible nonequimolar proportions determined by those in total cell extracts. These results show that virions differ in their genomic content but together build up a highly reproducible genetic composition of the viral population. This resembles the genome formula described for multipartite viruses, with which some species of the order Bunyavirales may share some aspects of the way of life, particularly emerging properties at a supravirion scale.

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Section: Discussionmentioning

confidence: 72%

The genome of a bunyavirus cannot be defined at the level of the viral particle but only at the scale of the viral population

Yvon,

German,

Ullman

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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“…NSP5 can then associate with VP1 ( 59 ) and TRiC (this study). VP6 is incorporated in the second layer to form the DLPs that bud to the ER to be converted into mature TLPs ( 83 ). As denoted by our results, the core-shell structure is preserved upon inhibition of TRiC and does not disturb the association of VP6, hence generating empty DLPs and TLPs.…”

Section: Discussionmentioning

confidence: 99%

The recruitment of TRiC chaperonin in rotavirus viroplasms correlates with virus replication

Vetter,

Papa,

Tobler

et al. 2024

mBio

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Rotavirus (RV) replication takes place in the viroplasms, cytosolic inclusions that allow the synthesis of virus genome segments and their encapsidation in the core shell, followed by the addition of the second layer of the virion. The viroplasms are composed of several viral proteins, including NSP5, which serves as the main building block. Microtubules, lipid droplets, and miRNA-7 are among the host components recruited in viroplasms. We investigated the interaction between RV proteins and host components of the viroplasms by performing a pull-down assay of lysates from RV-infected cells expressing NSP5-BiolD2. Subsequent tandem mass spectrometry identified all eight subunits of the tailless complex polypeptide I ring complex (TRiC), a cellular chaperonin responsible for folding at least 10% of the cytosolic proteins. Our confirmed findings reveal that TRiC is brought into viroplasms and wraps around newly formed double-layered particles. Chemical inhibition of TRiC and silencing of its subunits drastically reduced virus progeny production. Through direct RNA sequencing, we show that TRiC is critical for RV replication by controlling dsRNA genome segment synthesis, particularly negative-sense single-stranded RNA. Importantly, cryo-electron microscopy analysis shows that TRiC inhibition results in defective virus particles lacking genome segments and polymerase complex (VP1/VP3). Moreover, TRiC associates with VP2 and NSP5 but not with VP1. Also, VP2 is shown to be essential for recruiting TRiC in viroplasms and preserving their globular morphology. This study highlights the essential role of TRiC in viroplasm formation and in facilitating virion assembly during the RV life cycle. IMPORTANCE The replication of rotavirus takes place in cytosolic inclusions termed viroplasms. In these inclusions, the distinct 11 double-stranded RNA genome segments are co-packaged to complete a genome in newly generated virus particles. In this study, we show for the first time that the tailless complex polypeptide I ring complex (TRiC), a cellular chaperonin responsible for the folding of at least 10% of the cytosolic proteins, is a component of viroplasms and is required for the synthesis of the viral negative-sense single-stranded RNA. Specifically, TRiC associates with NSP5 and VP2, the cofactor involved in RNA replication. Our study adds a new component to the current model of rotavirus replication, where TRiC is recruited to viroplasms to assist replication.

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“…The mature RV virion is a nonenveloped, icosahedral (T = 13), triple-layered particle (TLP) [13] of about 100 nm in diameter [14]. The virion encapsidates one copy of each of the eleven double-stranded RNA (dsRNA) genome segments.…”

Section: Rotavirusmentioning

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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Rotavirus Particle Disassembly and Assembly In Vivo and In Vitro

Cited by 10 publications

References 184 publications

The genome of a bunyavirus cannot be defined at the level of the viral particle but only at the scale of the viral population

The genome of a bunyavirus cannot be defined at the level of the viral particle but only at the scale of the viral population

The recruitment of TRiC chaperonin in rotavirus viroplasms correlates with virus replication

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

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