Tracking the Fate of Genetically Distinct Vesicular Stomatitis Virus Matrix Proteins Highlights the Role for Late Domains in Assembly

Soh, Timothy K.; Whelan, Sean P. J.

doi:10.1128/jvi.01371-15

Cited by 24 publications

(19 citation statements)

References 48 publications

(52 reference statements)

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“…Whether this process is mediated by nuclear export activities of a viral nucleocapsid protein or via a budding mechanism awaits future studies. With the advent of plant rhabdovirus reverse genetics systems, it is now possible to tag viral proteins genetically with fluorescent proteins, as has been shown in several mammalian rhabdovirus studies (83)(84)(85)(86)(87)(88). This approach might permit intracellular and intercellular tracking of the movement of infectious virus entities in live cells.…”

Section: Discussionmentioning

confidence: 99%

Specificity of Plant Rhabdovirus Cell-to-Cell Movement

Zhou

Lin

Sun

et al. 2019

J Virol

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Positive-stranded RNA virus movement proteins (MPs) generally lack sequence-specific nucleic acid-binding activities and display cross-family movement complementarity with related and unrelated viruses. Negative-stranded RNA plant rhabdoviruses encode MPs with limited structural and functional relatedness with other plant virus counterparts, but the precise mechanisms of intercellular transport are obscure. In this study, we first analyzed the abilities of MPs encoded by five distinct rhabdoviruses to support cell-to-cell movement of two positive-stranded RNA viruses by using trans-complementation assays. Each of the five rhabdovirus MPs complemented the movement of MP-defective mutants of tomato mosaic virus and potato X virus. In contrast, movement of recombinant MP deletion mutants of sonchus yellow net nucleorhabdovirus (SYNV) and tomato yellow mottle-associated cytorhabdovirus (TYMaV) was rescued only by their corresponding MPs, i.e., SYNV sc4 and TYMaV P3. Subcellular fractionation analyses revealed that SYNV sc4 and TYMaV P3 were peripherally associated with cell membranes. A split-ubiquitin membrane yeast two-hybrid assay demonstrated specific interactions of the membraneassociated rhabdovirus MPs only with their cognate nucleoproteins (N) and phosphoproteins (P). More importantly, SYNV sc4-N and sc4-P interactions directed a proportion of the N-P complexes from nuclear sites of replication to punctate loci at the cell periphery that partially colocalized with the plasmodesmata. Our data show that cell-to-cell movement of plant rhabdoviruses is highly specific and suggest that cognate MP-nucleocapsid core protein interactions are required for intra-and intercellular trafficking. IMPORTANCE Local transport of plant rhabdoviruses likely involves the passage of viral nucleocapsids through MP-gated plasmodesmata, but the molecular mechanisms are not fully understood. We have conducted complementation assays with MPs encoded by five distinct rhabdoviruses to assess their movement specificity. Each of the rhabdovirus MPs complemented the movement of MP-defective mutants of two positive-stranded RNA viruses that have different movement strategies. In marked contrast, cell-to-cell movement of two recombinant plant rhabdoviruses was highly specific in requiring their cognate MPs. We have shown that these rhabdovirus MPs are localized to the cell periphery and associate with cellular membranes, and that they interact only with their cognate nucleocapsid core proteins. These interactions are able to redirect viral nucleocapsid core proteins from their sites of replication to the cell periphery. Our study provides a model for the specific interand intracellular trafficking of plant rhabdoviruses that may be applicable to other negative-stranded RNA viruses. Downloaded fromN atural infections of plant viruses are initiated from one or a few primary infected cells, followed by cell-to-cell spread to adjacent cells and then to distal tissues via vascular networks. To accomplish systemic infections, plant viruses must ci...

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Section: Discussionmentioning

confidence: 99%

Specificity of Plant Rhabdovirus Cell-to-Cell Movement

Zhou

Lin

Sun

et al. 2019

J Virol

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“…A functional fluorescent VSV M protein (MeGFP) was introduced into VSV-LUJV as reported. Fluorescent VSV-LUJV-MeGFP was amplified and purified as described previously (Soh and Whelan, 2015). VSV-LCMV, VSV-JUNV, VSV-GTOV and VSV-MACV were generated in a similar manner by using gBlocks Gene Fragments (from Integrated DNA Technologies) of the corresponding GP precursor coding sequences (GenBank accession number: AY847350.1 [for LCMV GP], GenBank accession number: D10072.2 [for JUNV GP], GenBank accession number: AF485258.1 [for GTOV GP] and GenBank accession number: KM198592.1 [for MACV GP]).…”

Section: Star ★ Methodsmentioning

confidence: 99%

NRP2 and CD63 Are Host Factors for Lujo Virus Cell Entry

Raaben

Jae

Herbert

et al. 2017

Cell Host & Microbe

118

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Summary Arenaviruses cause fatal haemorrhagic disease in humans. Old-World arenavirus glycoproteins (GPs) mainly engage α-dystroglycan as a cell surface receptor, while New-World arenaviruses hijack transferrin receptor. However, the Lujo virus (LUJV) glycoprotein does not cluster with New-or Old-world arenaviruses. Using a recombinant vesicular stomatitis virus containing LUJV glycoprotein (LUJV GP) as its sole attachment and fusion protein (VSV-LUJV), we demonstrate that infection is independent of known arenavirus receptor genes. A genome-wide haploid genetic screen identified the transmembrane protein neuropilin 2 (NRP2) and tetraspanin CD63 as factors for LUJV GP-mediated infection. LUJV GP binds the N-terminal domain of NRP2, while CD63 stimulates pH-activated LUJV GP-mediated membrane fusion. Overexpression of NRP2 or its N-terminal domain enhances VSV-LUJV infection, and cells lacking NRP2 are deficient in wild-type LUJV infection. These findings uncover this distinct set of host cell entry factors in LUJV infection and are attractive focus points for therapeutic intervention.

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“…Viral constructs containing nano-luciferase within the viral particle were made by replacing the matrix (M) gene with the M-nLuc construct. M-nLuc was generated by adding the nano-luciferase coding region after residue 37 as was previously done with GFP (55).…”

Section: Generation Of Replication Competent Recombinant Vsvmentioning

confidence: 99%

Ebola virus requires phosphatidylserine scrambling activity for efficient budding and optimal infectivity

Acciani

Lay-Mendoza

Havranek

et al. 2020

Preprint

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word count: 250 AbstractEbola virus (EBOV) interacts with cells using two categories of cell surface receptors, C-type lectins and phosphatidylserine (PS) receptors. PS receptors typically bind to apoptotic cell membrane PS and orchestrate the uptake and clearance of apoptotic bodies. Many viruses coated with PS-containing lipid envelopes, acquired during budding from host cells, can also exploit these receptors for internalization. PS is restricted to the inner leaflet of the plasma membrane in homeostatic cells, an orientation that would be unfavorable for PS receptor-mediated uptake if conserved on the viral envelope. Therefore, it is theorized that viral infection induces host cell PS externalization to the outer leaflet during replication. Cells have several membrane scramblase enzymes that enrich outer leaflet PS when activated. Here, we investigate two scramblases, TMEM16F and XKR8, as possible mediators of cellular and viral envelope surface PS levels during recombinant VSV/EBOV-GP replication and EBOV virus-like particle (VLP) production. We found that rVSV/EBOV-GP and EBOV VLPs produced in XKR8 knockout cells contain decreased levels of PS in their outer leaflets. ΔXKR8-made rVSV/EBOV-GP is 70% less efficient at infecting cells through apoptotic mimicry compared to viruses made in parental cells. Our data suggest that virion surface PS acquisition requires XKR8 activity, whereas TMEM16F activity is not essential. Unexpectedly, we observed defective rVSV/G, rVSV/EBOV-GP, and EBOV VLP budding in ΔXKR8 cells, suggesting that phospholipid scrambling via XKR8 enhances both Ebola infectivity and budding efficiency. Overexpression of XKR8 dramatically increased budding activity, suggesting outer leaflet PS is required for both particle production and increased infectivity. ImportanceThe Democratic Republic of the Congo experienced its deadliest Ebola outbreak from 2018 to 2020, with 3,444 confirmed cases and 2,264 deaths (as of March 12, 2020). Owing to the extensive damage that these outbreaks have caused in Africa, as well as its future epidemic potential, Ebola virus (EBOV) ranks among the top eight priority pathogens outlined by the WHO in 2018. A comprehensive understanding of Ebola entry pathways into target cells is critical for antiviral development and outbreak control. Thus far, host-cell scramblases TMEM16F and XKR8 have each been named as the sole mediator of Ebola envelope surface phosphatidylserine (PS). We assessed the contributions of these proteins using CRISPR knockout cells and two EBOV models: rVSV/EBOV-GP and EBOV VLPs. We observed that XKR8 is required for optimal EBOV envelope PS levels, PS receptor engagement, and particle budding across all viral models, whereas TMEM16F did not play a major role.

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Tracking the Fate of Genetically Distinct Vesicular Stomatitis Virus Matrix Proteins Highlights the Role for Late Domains in Assembly

Cited by 24 publications

References 48 publications

Specificity of Plant Rhabdovirus Cell-to-Cell Movement

Specificity of Plant Rhabdovirus Cell-to-Cell Movement

NRP2 and CD63 Are Host Factors for Lujo Virus Cell Entry

Ebola virus requires phosphatidylserine scrambling activity for efficient budding and optimal infectivity

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