Endoplasmic reticulum export and vesicle formation of the movement protein of Chinese wheat mosaic virus are regulated by two transmembrane domains and depend on the secretory pathway

Andika, Ida Bagus; Zheng, Shiling; Tan, Zilong; Sun, Liying; Kondō, Hideki; Zhou, Xueping; Chen, Jianping

doi:10.1016/j.virol.2012.10.024

Cited by 51 publications

(35 citation statements)

References 64 publications

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“…In the same manner, a (conservative) substitution of the equivalent Leu with Val or Ile resulted in improved TMV movement (Toth et al, 2002;Kawakami et al, 2003). In contrast, substitutions of the equivalent b1 residue abolished virus movement in TRV (this study), and in CPMV, Chinese wheat mosaic virus (CWMV) and Prunus necrotic ringspot virus (PNRSV), as well as tubule formation in CPMV (Bertens et al, 2000, Martínez-Gil et al, 2009Andika et al, 2013), whilst deletion of part of the b1 region prevented TMV movement (Fujiki et al, 2006).…”

Section: Mps Of the 30k Superfamilymentioning

confidence: 55%

Experimental and bioinformatic evidence that raspberry leaf blotch emaravirus P4 is a movement protein of the 30K superfamily

Chen

Karlin

et al. 2013

Journal of General Virology

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Emaravirus is a recently described genus of negative-strand RNA plant viruses. Emaravirus P4 protein localizes to plasmodesmata, suggesting that it could be a viral movement protein (MP). In the current study, we showed that the P4 protein of raspberry leaf blotch emaravirus (RLBV) rescued the cell-to-cell movement of a defective potato virus X (PVX) that had a deletion mutation in the triple gene block 1 movement-associated protein. This demonstrated that RLBV P4 is a functional MP. Sequence analyses revealed that P4 is a distant member of the 30K superfamily of MPs. All MPs of this family contain two highly conserved regions predicted to form b-strands, namely b1 and b2. We explored by alanine mutagenesis the role of two residues of P4 (Ile106 and Asp127) located in each of these strands. We also made the equivalent substitutions in the 29K MP of tobacco rattle virus, another member of the 30K superfamily. All substitutions abolished the ability to complement PVX movement, except for the I106A substitution in the b1 region of P4. This region has been shown to mediate membrane association of 30K MPs; our results show that it is possible to make non-conservative substitutions of a well-conserved aliphatic residue within b1 without preventing the membrane association or movement function of P4.

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Section: Mps Of the 30k Superfamilymentioning

confidence: 55%

Experimental and bioinformatic evidence that raspberry leaf blotch emaravirus P4 is a movement protein of the 30K superfamily

Chen

Karlin

et al. 2013

Journal of General Virology

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“…To construct split yellow fluorescent protein (YFP) plasmids for the bimolecular fluorescence complementation (BiFC) assay, we replaced the GFP gene in pBin.GFP with half-sized YFP fragments excluding the stop codon, YFPn (aa 1 to 155) or YFPc (aa 156 to 239). The resulting plasmids were termed pBin.YFPn and pBin.YFPc, respectively (27,28). The PCR fragments of the N, P, and P mutants were inserted into the BamHI and SalI sites of pBin.YFPn or pBin.YFPc to create pBin.…”

Section: Methodsmentioning

confidence: 99%

Orchid Fleck Virus Structural Proteins N and P Form Intranuclear Viroplasm-Like Structures in the Absence of Viral Infection

Kondō

Chiba

Andika

et al. 2013

J Virol

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bOrchid fleck virus (OFV) has a unique two-segmented negative-sense RNA genome that resembles that of plant nucleorhabdoviruses. In infected plant cells, OFV and nucleorhabdoviruses induce an intranuclear electron-lucent viroplasm that is believed to be the site for virus replication. In this study, we investigated the molecular mechanism by which OFV viroplasms are produced in vivo. Among OFV-encoded proteins, the nucleocapsid protein (N) and the putative phosphoprotein (P) were present in nuclear fractions of OFV-infected Nicotiana benthamiana plants. Transient coexpression of N and P, in the absence of virus infection, was shown to be sufficient for formation of an intranuclear viroplasm-like structure in plant cells. When expressed independently as a fluorescent protein fusion product in uninfected plant cells, N protein accumulated throughout the cell, while P protein accumulated in the nucleus. However, the N protein, when coexpressed with P, was recruited to a subnuclear region to induce a large viroplasm-like focus. Deletion and substitution mutagenesis demonstrated that the P protein contains a nuclear localization signal (NLS). Artificial nuclear targeting of the N-protein mutant was insufficient for formation of viroplasm-like structures in the absence of P. A bimolecular fluorescence complementation assay confirmed interactions between the N and P proteins within subnuclear viroplasm-like foci and interactions of two of the N. benthamiana importin-␣ homologues with the P protein but not with the N protein. Taken together, our results suggest that viroplasm formation by OFV requires nuclear accumulation of both the N and P proteins, which is mediated by P-NLS, unlike nucleorhabdovirus viroplasm utilizing the NLS on protein N. R habdoviruses (family Rhabdoviridae, order Mononegavirales) have a broad range of hosts, including humans, livestock, fish, plants, and insects (1). They have bacilliform or bulletshaped virions containing a nonsegmented negative-sense RNA [(Ϫ)RNA] as the genome (2). All of the well-characterized animal rhabdoviruses replicate in the cytoplasm (1). However, the plant rhabdoviruses are divided into two groups, depending on their sites of replication and morphogenesis. Members of the genus Cytorhabdovirus replicate and undergo morphogenesis in the cytoplasm, whereas members of the genus Nucleorhabdovirus multiply in plant cell nuclei and their virions bud through the inner nuclear envelopes (3, 4).Orchid fleck virus (OFV), a Brevipalpus (Acari: Tenuipalpidae) mite-transmitted virus, has a unique two-segmented (Ϫ)RNA genome that resembles that of plant rhabdoviruses (5, 6). OFV is distributed worldwide and infects many kinds of orchids (5,7,8). OFV RNA1 (GenBank accession number AB244417; 6,413 nucleotides [nt]) contains five genes in the order 3=-N-P-3-M-G-5=, in which N encodes the nucleocapsid protein (open reading frame 1 [ORF1]), P encodes the phosphoprotein (ORF2), gene 3 encodes the candidate movement protein (ORF3), M encodes the matrix protein (ORF4), and G encodes the can...

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“…Hence, plant viruses encode movement proteins (MPs) that facilitate trafficking through the host cell wall by altering the size exclusion limit (SEL) or the structure of plasmodesmata (Lucas 2006). PME binds to the MP of TMV, Turnip vein clearing virus (TVCV), Cauliflower mosaic virus and Chinese wheat mosaic virus, and this interaction is essential for cell-to-cell movement of TMV (Chen et al 2000;Andika et al 2013), Moreover, antisense suppression of PME, prevalently in the vasculature of transgenic tobacco plants causes a significant delay of the onset of TMV-GFP systemic movement due to a reduced phloem unloading of the virus. However, once started, TMV systemic movement proceeds in these plants with a rate comparable to that of wild-type plants (Chen and Citovsky 2003).…”

Section: Introductionmentioning

confidence: 99%

Virus-induced gene silencing of pectin methylesterase protects Nicotiana benthamiana from lethal symptoms caused by Tobacco mosaic virus

et al. 2014

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Plant viruses are obligate parasites that exploit host components for replication and spread inside the host. Transport of the viral genome is enabled by movement proteins (MPs) targeting the cell periphery to mediate passage throughout plasmodesmata (PD). Pectin methylesterase (PME) is one of the critical host factors facilitating MPs in PD gating, and a direct interaction of PME with Tobacco mosaic virus (TMV) MP is required for viral movement and in turn for virus viability. PME is a critical enzyme for host development and defence, acting via complex mechanisms involving multigenic and tissue specific isoforms and endogenous inhibitors. This composite activity of PME suggests that level and timing of protein accumulation, with respect to virus inoculation and MP expression, can be critical for the functional outcome of the PME-MP interaction and in turn for the success of a viral infection. Based on this notion, we tested different experimental conditions to evaluate the beneficial effect of the downregulation of PME gene expression on the development of TMVinduced disease and on plant protection. We used virus induced gene silencing technology (VIGS) to downregulate PME gene expression, which resulted in a 30-45 % reduction of TMV symptom severity and, correspondingly, to a 60 % reduction of TMV RNA accumulation in systemic leaves. VIGS proved to be a rapid and effective technology for PME gene silencing in functional assays and for plant defence from viral infection. Our findings indicate that N. benthamiana plants with hindered expression of PME survive a TMV infection, which kills non-silenced plants within a week.

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Endoplasmic reticulum export and vesicle formation of the movement protein of Chinese wheat mosaic virus are regulated by two transmembrane domains and depend on the secretory pathway

Cited by 51 publications

References 64 publications

Experimental and bioinformatic evidence that raspberry leaf blotch emaravirus P4 is a movement protein of the 30K superfamily

Experimental and bioinformatic evidence that raspberry leaf blotch emaravirus P4 is a movement protein of the 30K superfamily

Orchid Fleck Virus Structural Proteins N and P Form Intranuclear Viroplasm-Like Structures in the Absence of Viral Infection

Virus-induced gene silencing of pectin methylesterase protects Nicotiana benthamiana from lethal symptoms caused by Tobacco mosaic virus

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