Functional Analysis of a DNA-Shuffled Movement Protein Reveals That Microtubules Are Dispensable for the Cell-to-Cell Movement of <i>Tobacco mosaic virus</i>

Gillespie, Trudi; Boevink, Petra C.; Haupt, Sophie; Roberts, Alison G.; Toth, Rachel; Valentine, Tracy A.; Chapman, Sean; Oparka, Karl J.

doi:10.1105/tpc.002303

Cited by 160 publications

(188 citation statements)

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“…Tobacco mosaic virus (TMV), a virus with a (ϩ) single-stranded RNA genome encodes one of the most extensively studied movement proteins, MP30. Several biochemical and cell biological features of MP30 were revealed: MP30 binds cooperatively single-stranded nucleic acids (Citovsky et al, 1990), interacts with the tubulin-and actin-based cytoskeleton (Heinlein et al, 1995;McLean et al, 1995), associates to the endoplasmic reticulum (ER; Heinlein et al, 1998;Reichel and Beachy, 1998;Mas and Beachy, 1999;Gillespie et al, 2002), and increases the size exclusion limit of plasmodesmata (Wolf et al, 1989;Waigmann et al, 1994). In the current model, MP30 is proposed to form complexes with genomic TMV-RNA (vRNA) and to move these vRNA-protein complexes from ER-associated sites of synthesis throughout the cell via the cytoskeletal network toward plasmodesmata.…”

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confidence: 99%

MPB2C, a Microtubule-Associated Plant Protein Binds to and Interferes with Cell-to-Cell Transport of Tobacco Mosaic Virus Movement Protein

et al. 2003

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The movement protein of tobacco mosaic virus, MP30, mediates viral cell-to-cell transport via plasmodesmata. The complex MP30 intra-and intercellular distribution pattern includes localization to the endoplasmic reticulum, cytoplasmic bodies, microtubules, and plasmodesmata and likely requires interaction with plant endogenous factors. We have identified and analyzed an MP30-interacting protein, MPB2C, from the host plant Nicotiana tabacum. MPB2C constitutes a previously uncharacterized microtubule-associated protein that binds to and colocalizes with MP30 at microtubules. In vivo studies indicate that MPB2C mediates accumulation of MP30 at microtubules and interferes with MP30 cell-to-cell movement. In contrast, intercellular transport of a functionally enhanced MP30 mutant, which does not accumulate and colocalize with MP30 at microtubules, is not impaired by MPB2C. Together, these data support the concept that MPB2C is not required for MP30 cell-to-cell movement but may act as a negative effector of MP30 cell-to-cell transport activity.Plant viruses produce movement proteins required to mediate cell-to-cell spread of the viral genomic information via plasmodesmata (Lucas, 1995;Lazarowitz and Beachy, 1999;McLean and Zambryski, 2000;Tzfira et al., 2000;Haywood et al., 2002;Heinlein, 2002). Tobacco mosaic virus (TMV), a virus with a (ϩ) single-stranded RNA genome encodes one of the most extensively studied movement proteins, MP30. Several biochemical and cell biological features of MP30 were revealed: MP30 binds cooperatively single-stranded nucleic acids (Citovsky et al., 1990), interacts with the tubulin-and actin-based cytoskeleton (Heinlein et al., 1995;McLean et al., 1995), associates to the endoplasmic reticulum (ER; Heinlein et al., 1998;Reichel and Beachy, 1998;Mas and Beachy, 1999;Gillespie et al., 2002), and increases the size exclusion limit of plasmodesmata (Wolf et al., 1989;Waigmann et al., 1994). In the current model, MP30 is proposed to form complexes with genomic TMV-RNA (vRNA) and to move these vRNA-protein complexes from ER-associated sites of synthesis throughout the cell via the cytoskeletal network toward plasmodesmata. There, MP30 induces an increase in plasmodesmal size exclusion limit and facilitates the transport of the vRNA-MP30 complex through the enlarged plasmodesmal channels into adjacent cells (Lazarowitz and Beachy, 1999;Tzfira et al., 2000;Haywood et al., 2002;Heinlein, 2002). In line with this model, microtubules have been functionally implicated in guiding vRNA-MP30 complexes toward plasmodesmata (Mas and Beachy, 2000; Boyko et al., 2000a Boyko et al., , 2000b. However, an alternate, non-movement-promoting role of microtubules in providing a route toward an MP30 degradation site was suggested (Padgett et al., 1996;Mas and Beachy, 1999;Reichel and Beachy, 2000). This model is supported by the observation that TMV was able to spread in tissues with functionally impaired microtubules and by studies performed with a mutant variant of MP30, MP R3 with strongly decreased affinity...

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confidence: 99%

MPB2C, a Microtubule-Associated Plant Protein Binds to and Interferes with Cell-to-Cell Transport of Tobacco Mosaic Virus Movement Protein

et al. 2003

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“…Several studies (Wright et al, 2007;GuenouneGelbart et al, 2008;Epel, 2009) support a model in which targeting of MP to PD during infection is mediated by the actin/ER network in the absence of an intact microtubule (MT) cytoskeleton. Some reports suggest that the MT cytoskeleton is involved in the degradation pathway of MP (Gillespie et al, 2002;Mas & Beachy, 1999;. On the other hand, as has been reported by Ashby et al (2006), MT-associated MP is not ubiquitinated, and the ER/actin network may act together with MT in intracellular trafficking of MP-containing RNA transport particles (Sambade et al, 2008).…”

Section: Introductionmentioning

confidence: 79%

Binding of monoclonal antibodies to the movement protein (MP) of Tobacco mosaic virus: influence of subcellular MP localization and phosphorylation

Tyulkina

Karger

Sheveleva

et al. 2010

Journal of General Virology

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Monoclonal antibodies (mAbs) to recombinant movement protein (MP REC ) of Tobacco mosaic virus (TMV) were used to reveal the dependence of MP epitope accessibility to mAbs on subcellular MP localization and post-translational MP phosphorylation. Leaves of Nicotiana benthamiana or N. tabacum were inoculated mechanically with TMV or agroinjected with an MP expression vector. At different time post-inoculation, ER membrane-and cell wall-enriched fractions (ER-MP and CW-MP, respectively) were isolated and analysed. The N-terminal region (residues 1-30) as well as regions 186-222 and 223-257 of MP from the CW and ER fractions were accessible for interaction with mAbs. By contrast, the MP regions including residues 76-89 and 98-129 were not accessible. The C-terminal TMV MP region (residues 258-268) was inaccessible to mAbs not only in CW-MP, but also in ER-MP fractions. Evidence is presented that phosphorylation of the majority of TMV MP C-terminal sites occurred on ER membranes at an early stage of virus infection, i.e. not after, but before reaching the cell wall. C-terminal phosphorylation of purified MP REC abolished recognition of C-proximal residues 258-268 by specific mAbs, which could be restored by MP dephosphorylation. Likewise, accessibility to mAbs of the C-terminal MP epitope in ER-MP and CW-MP leaf fractions was restored by dephosphorylation. Substitution of three or four C-terminal Ser/Thr residues with nonphosphorylatable Ala also resulted in abolition of interaction of mAbs with MP. INTRODUCTIONThe movement protein (MP) of Tobacco mosaic virus (TMV) facilitates movement of the virus genome within and between cells (for reviews, see Citovsky, 1999;Heinlein & Epel, 2004;Lucas, 2006). Polyfunctional TMV MP plays a key role in virus infection. It increases the size exclusion limit of plasmodesmata (PD) (Tomenius et al., 1987;Deom et al., 1987;Wolf et al., 1989), serves as an anchor of the viral genomic RNA to the ER (Mas & Beachy, 1999) and plays an important role in intracellular transport of replication complexes Kawakami et al., 2004). Unlike other TMV proteins, the MP is produced transiently in the early stages of virus infection (Watanabe et al., 1984;Lehto et al., 1990). Within the infected cell, MP is synthesized at the leading edge and degraded at the trailing edge of the infection site (Padgett et al., 1996). Polyubiquitination of MP and subsequent degradation by the 26S proteasome pathway occurs after the early stages of infection (Reichel & Beachy, 2000). However, the mechanisms responsible for the regulation of transient MP expression are not clear.Post-translational modifications, in particular phosphorylation/dephosphorylation, are important in regulation of gene expression and protein activity. It is known that, during virus infection, TMV MP undergoes phosphorylation by a cellular protein kinase(s). It has been shown that multiple phosphorylation events in internal regions of TMV MP occur in TMV-infected protoplasts (Haley et al., 1995). On the other hand, only three C-terminal residues o...

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“…I-class b-1,3-glucanase limits callose accumulation near PD which is induced by viral infection due to the diffusion of MP-vRNA-replication complex in ER-desmotubules membranes (Fig. 1) (Gillespie et al 2002;Kawakami et al 2004;Liu et al 2005). GuenouneGelbart et al (2008) suggested that the cytoskeleton is not directly involved.…”

Section: Mp Tmv and Its Partners In Movementmentioning

confidence: 99%

“…Kragler et al (2003) stressed the specific interaction between MP TMV and microtubule associated protein in tobacco tissues-MPB2C (movement protein binding 2C). This endogenous plant agent may play a role of a negative effector in MP TMV cell-to-cell movement (Gillespie et al 2002). Waigmann et al (2004) postulated only indirect involvement of MPB2C in TMV cell-to-cell transport.…”

Section: Mp Tmv and Its Partners In Movementmentioning

confidence: 99%

“…Actins and myosins are present inside PD and are responsible for the ER membrane translocation (Blackman and Overall 1998;Yokota et al 2009). Gillespie et al (2002) and Heinlein and Epel (2004) postulated that PD broadening in induced MP may enable an active transport of vesicles from ER (with MP-vRNA) due to microfilaments. It was estimated that cell-to-cell transport of the vesicles (with virus complex) with the cytoskeleton involved in I-stage infection cells reaches 160 nm/s (Kawakami et al 2004).…”

Section: Mp Tmv and Its Partners In Movementmentioning

confidence: 99%

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Cell-to-cell movement of three genera (+) ss RNA plant viruses

Otulak

Garbaczewska

2010

Acta Physiol Plant

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The current investigations of three genera plant virus cell-to-cell movement were presented. Viruses reveal different local transport strategies, but all of them are the results of virus factors-host components interactions. The Tobacco mosaic virus (TMV) does not require capsid protein for translocation through plasmodesmata but 30 K movement protein participates in this process. It was found direct or indirect TMV movement proteins host partners in Tobamovirus movement like: pectin methylesterase, movement protein binding 2C, chaperones or cytoskeleton components and endoplasmatic reticulum membranes. The Potex-and Potyvirus cell-to-cell movement is closely related to replication network. The PVX capsid protein and triple gene block protein system are responsible for efficient local transport. Potyviruses move through the plasmodesmata by involving viral encoded proteins but not specific movement proteins. While the Potyvirus is the biggest known plant virus genus, host components participating in or regulating directly its plasmodesmata-movement are still not clear.

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Functional Analysis of a DNA-Shuffled Movement Protein Reveals That Microtubules Are Dispensable for the Cell-to-Cell Movement of Tobacco mosaic virus

Abstract: Microtubules interact strongly with the viral movement protein (MP) of

Cited by 160 publications

References 61 publications

MPB2C, a Microtubule-Associated Plant Protein Binds to and Interferes with Cell-to-Cell Transport of Tobacco Mosaic Virus Movement Protein

MPB2C, a Microtubule-Associated Plant Protein Binds to and Interferes with Cell-to-Cell Transport of Tobacco Mosaic Virus Movement Protein

Binding of monoclonal antibodies to the movement protein (MP) of Tobacco mosaic virus: influence of subcellular MP localization and phosphorylation

Cell-to-cell movement of three genera (+) ss RNA plant viruses

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