The role of signalling and the cytoskeleton during Vaccinia Virus egress

Leite, Flávia G. G.; Way, Michael

doi:10.1016/j.virusres.2015.01.024

Cited by 44 publications

(59 citation statements)

References 137 publications

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“…The cell cytoskeleton is known to participate in viral infection in vaccinia virus (35). As reported previously (32)(33)(34) and quantitated by our IFC analysis, cells infected with Mimivirus show a cytopathic effect, undergoing dramatic morphological changes likely to involve cytoskeleton rearrangements.…”

Section: Mimivirus Infection Requires Certain Host Cytoskeleton Filamsupporting

confidence: 77%

Kinetics of Mimivirus Infection Stages Quantified Using Image Flow Cytometry

et al. 2019

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Due to the heterogeneity of viruses and their hosts, a comprehensive view of viral infection is best achieved by analyzing large populations of infected cells. However, information regarding variation in infected cell populations is lost in bulk measurements. Motivated by an interest in the temporal progression of events in virally infected cells, we used image flow cytometry (IFC) to monitor changes in Acanthamoeba polyphaga cells infected with Mimivirus . This first use of IFC to study viral infection required the development of methods to preserve morphological features of adherent amoeba cells prior to detachment and analysis in suspension. It also required the identification of IFC parameters that best report on key events in the Mimivirus infection cycle. The optimized IFC protocol enabled the simultaneous monitoring of diverse processes including generation of viral factories, transport, and fusion of replication centers within the cell, accumulation of viral progeny, and changes in cell morphology for tens of thousands of cells. After obtaining the time windows for these processes, we used IFC to evaluate the effects of perturbations such as oxidative stress and cytoskeletal disruptors on viral infection. Accurate dose‐response curves could be generated, and we found that mild oxidative stress delayed multiple stages of virus production, but eventually infection processes occurred with approximately the same amplitudes. We also found that functional actin cytoskeleton is required for fusion of viral replication centers and later for the production of viral progeny. Through this report, we demonstrate that IFC offers a quantitative, high‐throughput, and highly robust approach to study viral infection cycles and virus–host interactions. © The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

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Section: Mimivirus Infection Requires Certain Host Cytoskeleton Filamsupporting

confidence: 77%

Kinetics of Mimivirus Infection Stages Quantified Using Image Flow Cytometry

et al. 2019

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“…Vaccinia virus, for example, replicates its genome and assembles viral particles in the cytoplasm of cells, and then it uses kinesin-dependent transport to shuttle these viral particles to the plasma membrane for release from the cell (Leite and Way 2015). The timing of viral particle association with kinesin is regulated, in part, by the timing of viral gene expression to ensure that only mature viral particles are transported to the cell periphery.…”

Section: Fundamentals Of Microtubule-based Transportmentioning

confidence: 99%

Microtubule-Based Transport and the Distribution, Tethering, and Organization of Organelles

Barlan

Gelfand

2017

Cold Spring Harb Perspect Biol

173

157

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SUMMARY Microtubules provide long tracks along which a broad range of organelles and vesicles are transported by kinesin and dynein motors. Motor protein complexes also tether cargoes to cytoskeletal filaments, helping facilitate their interaction and communication. The generation of biochemically distinct microtubule subpopulations allows subsets of motors to recognize a given microtubule identity, allowing further organization within the cytoplasm. Both transport and tethering are spatiotemporally regulated through multiple modes, including acute modification of both motor–cargo and motor–track associations by various physiological signals. Strict regulation of intracellular transport is particularly important in specialized cell types such as neurons. Here, we review general mechanisms by which cargo transport is controlled and also highlight examples of transport regulated by multiple mechanisms.

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“…Deletion of the A36R gene and loss of A36 expression does not affect IEV assembly, but they do lead to an absence of actin tail formation and a small plaque size phenotype, indicative of a defect in viral spread (9, 10, 13). The phosphorylation of Tyr112 and Tyr132 in A36 by Src and Abl family kinases results in the recruitment of the adaptor proteins Nck, WIP, N-WASP, and Grb2, which in turn leads to activation of the Arp2/3 complex and the nucleation of actin polymerization (11,12,14). Mutations of Tyr112 and Tyr132 result in a loss of actin tail formation and in a reduction in plaque size (15).…”

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confidence: 99%

“…The mechanism of VACV actin tail formation has been intensively investigated with different VACV viruses (11), but not with NYVAC. Although the A33 and A36 transmembrane proteins are both required for actin tail formation, only A36 has been shown to have a direct role in this process (9)(10)(11)(12). A36R is highly conserved in Orthopoxvirus genomes, suggesting that virus-induced actin polymerization at the plasma membrane is widely used by mammalian poxviruses to enhance their cell-to-cell spread (11).…”

mentioning

confidence: 99%

“…Although the A33 and A36 transmembrane proteins are both required for actin tail formation, only A36 has been shown to have a direct role in this process (9)(10)(11)(12). A36R is highly conserved in Orthopoxvirus genomes, suggesting that virus-induced actin polymerization at the plasma membrane is widely used by mammalian poxviruses to enhance their cell-to-cell spread (11). Deletion of the A36R gene and loss of A36 expression does not affect IEV assembly, but they do lead to an absence of actin tail formation and a small plaque size phenotype, indicative of a defect in viral spread (9,10,13).…”

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confidence: 99%

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Suppression of NYVAC Infection in HeLa Cells Requires RNase L but Is Independent of Protein Kinase R Activity

Fernández‐Escobar

Nájera

Baldanta

et al. 2016

J Virol

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c Protein kinase R (PKR) and RNase L are host cell components that function to contain viral spread after infections. In this study, we analyzed the role of both proteins in the abortive infection of human HeLa cells with the poxvirus strain NYVAC, for which an inhibition of viral A27L and B5R gene expression is described. Specifically, the translation of these viral genes is independent of PKR activation, but their expression is dependent on the RNase L activity. NYVAC is a vaccinia virus (VACV) strain that is replication incompetent in most human cells and is currently being used as a recombinant poxvirus vaccine vector against multiple diseases (for a review, see reference 1). One of the advantages of replication-deficient viruses is their safety profile, which makes them excellent vehicles for vaccination purposes. However, it has been postulated that the efficacy of replication-incompetent viruses, like NYVAC, is limited by their failure to replicate and the consequent limitation in antigen accumulation during virus infection (1). It has been described that during the course of NYVAC infection in human HeLa cells, there is a late translational blockage that correlates with a marked increase in apoptosis (2, 3). An increase in the phosphorylation status of the translation initiation factor eIF2␣ (the ␣ subunit of eukaryotic initiation factor 2) is associated with this inhibition of protein synthesis during NYVAC infection. In particular, late viral proteins such as those encoded by A27L (A27 protein), A17L (A17 protein), B5R (B5 protein), and L1R (L1 protein) genes are not detected in HeLa cells infected with NYVAC, while other non-late viral proteins, such as those encoded by E3L (E3 protein) or A4L (A4 protein) or the early and late A36R (A36 protein) open reading frames (ORFs) are synthesized (2, 3). To understand what leads to the lack of these proteins, we have analyzed which step in the viral life cycle is blocked in NYVAC-infected HeLa cells. We compared viral protein synthesis in HeLa cells infected with either NYVAC or the replication-competent WR VACV strain, using Western blot analysis with specific antibodies for some early (E3 and A36) and late (B5 and A27) viral proteins. As shown in Fig. 1A, the early proteins E3 and A36 were detected in both WR-and NYVAC-infected cells, and their expression was maintained throughout the infection. In contrast, the late proteins B5 and A27 were only detected in WR-infected HeLa cells, indicating a block in their expression during NYVAC infection. The levels of early viral proteins were quite similar with both viruses at 2 h postinfection (hpi), but with longer times postinection, the levels of E3 and A36 were diminished in NYVACinfected cells due to the severe blockage in protein translation due to phosphorylation of the initiation factor eIF2␣, as previously published (2, 3). These results were confirmed by immunofluorescence analysis (data not shown) and are consistent with previous results obtained in human dendritic cells (DCs) and macrophages infected wit...

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The role of signalling and the cytoskeleton during Vaccinia Virus egress

Cited by 44 publications

References 137 publications

Kinetics of Mimivirus Infection Stages Quantified Using Image Flow Cytometry

Kinetics of Mimivirus Infection Stages Quantified Using Image Flow Cytometry

Microtubule-Based Transport and the Distribution, Tethering, and Organization of Organelles

Suppression of NYVAC Infection in HeLa Cells Requires RNase L but Is Independent of Protein Kinase R Activity

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