Association of bluetongue virus with the cytoskeleton

Eaton, Bryan T.; Hyatt, Alex D.; White, John R.

doi:10.1016/0042-6822(87)90319-9

Cited by 84 publications

(48 citation statements)

References 21 publications

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“…This result is indicative of the ability of AHSV CLPs to associate with the cytoskeleton in the absence of the other viral proteins. The prototypical orbivirus virion was also reported to be attached to the cytoskeleton singly or in such arrays (Eaton et al, 1987). The production of CLPs that structurally resembled empty AHSV cores and bound to AHSV VP7 monoclonal antibodies, was demonstrated.…”

Section: Discussionmentioning

confidence: 93%

Synthesis of empty african horse sickness virus particles

Maree

Putterill

et al. 2016

Virus Research

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As a means to develop African horse sickness (AHS) vaccines that are safe and DIVA compliant, we investigated the synthesis of empty African horse sickness virus (AHSV) particles. The emphasis of this study was on the assembly of the major viral core (VP3 and VP7) and outer capsid proteins (VP2 and VP5) into architecturally complex, heteromultimeric nanosized particles. The production of fully assembled corelike particles (CLPs) was accomplished in vivo by baculovirus-mediated co-synthesis of VP3 and VP7. The two different outer capsid proteins were capable of associating independently of each other with preformed cores to yield partial virus-like particles (VLPs). Complete VLPs were synthesized, albeit with a low yield. Crystalline formation of AHSV VP7 trimers is thought to impede high-level CLP production. Consequently, we engineered and co-synthesized VP3 with a more hydrophilic mutant VP7, resulting in an increase in the turnover of CLPs.

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Section: Discussionmentioning

confidence: 93%

Synthesis of empty african horse sickness virus particles

Maree

Putterill

et al. 2016

Virus Research

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“…The prototype of the Reoviridae family, reovirus needs an intact microtubule network to form their inclusion bodies (Dales 1963, Parker et al 2002, and other members like bluetongue viruses form their inclusion bodies associated to intermediate filaments (Eaton et al 1987). On the other hand, the viroplasms of rotavirus are formed by numerous viral proteins and RNAs, however NSP2 and NSP5 seem to be their elementary components, since these proteins are sufficient to form VLS when co-expressed in the absence of other viral components (Fabretti et al 1999).…”

Section: Discussionmentioning

confidence: 99%

“…The prototype of the Reoviridae family is the genus Reovirus that requires an intact microtubule network for virus assembly (Dales 1963, Parker et al 2002. Likewise, members of the genus Orbivirus within the Reoviridae form viral inclusion bodies associated to intermediate filaments (Eaton et al 1987). However, the role of the cytoskeleton in rotavirus viroplasms formation is unknown.…”

mentioning

confidence: 99%

Association of rotavirus viroplasms with microtubules through NSP2 and NSP5

Cabral-Romero

Padilla-Noriega

2006

Mem. Inst. Oswaldo Cruz

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Rotavirus replication and virus assembly take place in electrodense spherical structures known as viroplasms whose main components are the viral proteins NSP2 and NSP5. The viroplasms are produced since early times after infection and seem to grow by stepwise addition of viral proteins and by fusion, however, the mechanism of viropIasms formation is unknown. In this study we found that the viroplasms surface colocalized with microtubules, and seem to be caged by a microtubule network. Moreover inhibition of microtubule assembly with nocodazole interfered with viroplasms growth in rotavirus infected cells. We searched for a physical link between viroplasms and microtubules by co-immunoprecipitation assays, and we found that the proteins NSP2 and NSP5 were co-immunoprecipitated with anti-tubulin in rotavirus infected cells and also when they were transiently co-expressed or individually expressed. These results indicate that a functional microtubule network is needed for viroplasm growth presumably due to the association of viroplasms with microtubules via NSP2 and NSP5.Key words: rotavirus -viroplasms -microtubules -NSP2 -NSP5Group A rotaviruses are the leading etiological agents of severe diarrheal disease in infants and young children worldwide (Parashar 2003). These viruses belong to the genus Rotavirus within the Reoviridae family and their genome consist of 11 double-stranded RNA segments that encode six structural proteins named VP1-VP4, VP6, and VP7, as well as six nonstructural proteins named NSP1 to NSP6 (Estes 2001).Rotavirus replication takes place in the cytoplasm of infected cells in electrodense spherical structures known as "viroplasms" that can be found as early as 4 h after infection . Viroplasms are composed of viral RNA and the proteins VP1, VP2, VP3, VP6, NSP2, and NSP5 (Petrie et al. 1982, Gallegos & Patton 1989, and while virion assembly occur in these structures (Petrie et al. 1982, Gallegos & Patton 1989) the mechanism of viroplasms formation is unknown. Recently, it was reported that the number of rotavirus viroplasms decrease with post-infection time (Eichwald et al. 2004), and while the diameter of single viroplasms increased with time, the total number of viroplasms per cell diminishes, suggesting that growth of these inclusion bodies occur by fusion and probably also by stepwise addition of viral components to the viroplasms surface (Eichwald et al. 2004).In the viroplasms the viral mRNAs are replicated to produce genomic double stranded RNAs (dsRNA) which are simultaneously encapsidated into double layerded viral particles (DLPs), that contain the 11 dsRNA at the core of the particle surrounded by the inner and intermediate protein layers of VP2 and VP6, respectively. The third layer formed by VP4 and VP7 is acquired by budding of the DLP particles into the endoplasmic reticulum (ER), a process that requires assistance of the viral non-structural protein 4 (NSP4), whereby a transient envelope is acquired, that is finally lost within the ER along with NSP4. The fundamental role of NSP...

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“…per cell and replication was stopped at 2, 4, 6, 8, 10, 12, 16, 20, 24 and 28 h post-infection (p.i.). The monolayers were processed as described by Eaton et al (1987) with the exception that they were removed as ' floating sheets' using n-butyl glycidyl ether (Arnold & Boor, 1986). All specimens were examined with a Hitachi scanning transmission electron microscope the magnification of which was calibrated using a 2160 lines/mm diffracting grating replica standard.…”

mentioning

confidence: 99%

“…Monolayers of SVP cells (Eaton et al, 1987) were infected with BTV-1 (Australia) at an m.o.i, of 10 p.f.u. per cell and replication was stopped at 2, 4, 6, 8, 10, 12, 16, 20, 24 and 28 h post-infection (p.i.).…”

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confidence: 99%

Characterization of virus inclusion bodies in bluetongue virus-infected cells

Brookes

Hyatt

Eaton

1993

Journal of General Virology

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A combined qualitative and quantitative approach has been used to examine the role of virus inclusion bodies (VIBs) in the morphogenesis ofbluetongue virus (BTV).VIBs were detected as early as 4h post-infection (p.i.), and their number and profile areas increased significantly between 12 and 16 h, and 20 and 28 h p.i.respectively. Core-and virus-like particles were found within and at the periphery of the VIB matrix, respectively, and their numerical density (number per area of VIB matrix) decreased during the course of infection whereas the numerical density of virus particles in the cytoplasm increased. Virus-like particles had a diameter of 57 + 8 nm and core-like particles appeared to fall into two size ranges, 32+3 nm and 38+__3 nm in diameter. Both pre-and post-embedding immunoelectron microscopy procedures were used to localize BTV structural and non-structural proteins within the VIBs. The VIB matrix was labelled with antibodies to structural proteins VP5 and VP7 and non-structural proteins NS1 and NS2. Cores within VIBs contained proteins VP5, VP7 and NS1 but not VP2. Virus-like particles at the periphery of VIBs contained VP2, VP5, VP7 and NS1. The results suggest that BTV particles are synthesized, assembled and released from the perimeter of VIBs and not from within the matrix. Cores embedded in the VIBs are likely to have been trapped there during expansion of the matrix during replication.

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Association of bluetongue virus with the cytoskeleton

Cited by 84 publications

References 21 publications

Synthesis of empty african horse sickness virus particles

Synthesis of empty african horse sickness virus particles

Association of rotavirus viroplasms with microtubules through NSP2 and NSP5

Characterization of virus inclusion bodies in bluetongue virus-infected cells

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