Evidence for the Involvement of the 58K and 48K Proteins in the Intercellular Movement of Cowpea Mosaic Virus

Lent, J. van; Wellink, J.; Goldbach, Rob

doi:10.1099/0022-1317-71-1-219

Cited by 136 publications

(80 citation statements)

References 21 publications

(19 reference statements)

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“…Furthermore, immunofluorescence studies on the location of the 58K/48K movement proteins show that these proteins are also present over the whole length of the tubule. These results agree with our present and earlier observations from immunogold labelling of ultrathin sections of protoplasts and plant tissues (van Lent et al, 1990b). The exact location of the movement proteins could not be determined by labelling of negatively stained preparations because tubules were only labelled with antibodies against the 58K/48K proteins when the structure of the tubule had apparently disintegrated or was absent.…”

Section: Discussionsupporting

confidence: 83%

“…Recently, we have shown that the 58K/48K movement proteins of CPMV are localized to the tubular structures extending from plasmodesmata of infected cells, suggesting that these structures are involved in cell-to-cell movement (van Lent et al, 1990b). The limited ultrastructure detectable in ultrathin sections of tissue embedded at low temperature and the poor resolution of immunogold labelling make it difficult to determine the exact location of proteins.…”

Section: Introductionmentioning

confidence: 99%

“…Cowpea plants were inoculated with CPMV by differential temperature treatment (DTT; Dawson & Schlegel, 1976), and 24 and 48 h later leaf samples were processed for electron microscopy as described previously (van Lent et al, 1990b). Protoplasts were fixed, dehydrated and embedded in LR Gold at 12, 21, 29 and 40 h post-inoculation (p.i.…”

Section: Methodsmentioning

confidence: 99%

“…It has been shown previously that the overlapping 58K and 48K movement proteins of CPMV are associated with the tubular structures (van Lent et al, 1990b), implicating these structures in cell-to-cell movement. As it was assumed that the tubules were of viral origin we investigated whether they were also formed in CPMVinfected cowpea protoplasts in the absence of cell walls and, consequently, in the absence of plasmodesmata.…”

Section: Ultrastructure O F Virus-containing Tubules In Cowpea Plant mentioning

confidence: 99%

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Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts

Lent¹,

Storms²,

Meer³

et al. 1991

Journal of General Virology

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166

104

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In cowpea plant cells infected with cowpea mosaic virus, tubular structures containing virus particles are formed in the plasmodesmata between adjacent cells; these structures are supposedly involved in cell-to-cell spread of the virus. Here we show that similar tubular structures are also formed in cowpea protoplasts, from which the cell wall and plasmodesmata are absent. Between 12 and 21 h post-inoculation, tubule formation starts in the periphery of the protoplast at the level of the plasma membrane. Upon assembly, the viruscontaining tubule is enveloped by the plasma membrane and extends into the culture medium. This suggests that the tubule has functional polarity and makes it likely that a tubule 'grows' into a neighbouring cell in vivo. On average, 75 % of infected protoplasts were shown to possess tubular structures extending from their surface. The tubule wall was 3 to 4 nm thick and they were up to 20 gm in length, as shown by fluorescent light microscopy and negative staining electron microscopy. By analogy to infected plant cells, both the viral 58K/48K movement and capsid proteins were located in these tubules, as determined by immunofluorescent staining and immunogold labelling using specific antisera against these proteins. These results demonstrate that the formation of tubules is not necessarily dependent on the presence of plasmodesmata or the cell wall, and that they are composed, at least in part, of virus-encoded components.

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Section: Discussionsupporting

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Ultrastructure O F Virus-containing Tubules In Cowpea Plant mentioning

confidence: 99%

See 2 more Smart Citations

Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts

Lent¹,

Storms²,

Meer³

et al. 1991

Journal of General Virology

Self Cite

166

104

View full text Add to dashboard Cite

show abstract

“…Immunogold labelling has shown that the CPMV MP is present in these tubular structures (van Lent et al, 1990). On the surface of CPMV-infected protoplasts, similar tubular structures are formed, which protrude up to 20 mm into the culture medium, are tightly surrounded by the plasma membrane and have the same ultrastructure as tubules in plant tissue (van Lent et al, 1991).…”

Section: Introductionmentioning

confidence: 86%

Studies on the origin and structure of tubules made by the movement protein of Cowpea mosaic virus

Pouwels¹,

Velden²,

Willemse³

et al. 2004

Journal of General Virology

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Cowpea mosaic virus (CPMV) moves from cell to cell by transporting virus particles via tubules formed through plasmodesmata by the movement protein (MP). On the surface of protoplasts, a fusion between the MP and the green fluorescent protein forms similar tubules and peripheral punctate spots. Here it was shown by time-lapse microscopy that tubules can grow out from a subset of these peripheral punctate spots, which are dynamic structures that seem anchored to the plasma membrane. Fluorescence resonance energy transfer experiments showed that MP subunits interacted within the tubule, where they were virtually immobile, confirming that tubules consist of a highly organized MP multimer. Fluorescence recovery after photobleaching experiments with protoplasts, transiently expressing fluorescent plasma membrane-associated proteins of different sizes, indicated that tubules made by CPMV MP do not interact directly with the surrounding plasma membrane. These experiments indicated an indirect interaction between the tubule and the surrounding plasma membrane, possibly via a host plasma membrane protein. INTRODUCTIONFor successful systemic infection, a plant virus must spread throughout the plant, a process that starts with transport from the initially infected cell to neighbouring uninfected cells (cell-to-cell movement) and is followed by transport through the phloem into roots and young developing leaves (systemic movement). Since plant viruses cannot pass through the rigid cell wall, they have evolved ways of exploiting plasmodesmata, the naturally occurring transport channels present between plant cells (Haywood et al., 2002). Normally, only small molecules are able to pass through plasmodesmata, but plant viruses encode one or more proteins, the so-called movement proteins (MPs), that modify the structure of plasmodesmata in such a way that viral transport is enabled. So far, two basic principles for cell-to-cell movement of plant viruses have been described: tubule-guided movement of virus particles and movement as ribonucleoprotein complexes (Lazarowitz & Beachy, 1999).Cowpea mosaic virus (CPMV), a positive-stranded, bipartite RNA virus belonging to the family Comoviridae, moves from cell to cell by transporting virus particles using tubular structures, which connect the infected cell to the neighbouring uninfected cell (Pouwels et al., 2002a). Immunogold labelling has shown that the CPMV MP is present in these tubular structures (van Lent et al., 1990). On the surface of CPMV-infected protoplasts, similar tubular structures are formed, which protrude up to 20 mm into the culture medium, are tightly surrounded by the plasma membrane and have the same ultrastructure as tubules in plant tissue (van Lent et al., 1991). Remarkably, tubules are also formed on protoplasts transiently expressing MP , showing that MP is the only viral protein required for tubule formation. So far, protoplasts have proved extremely useful as a model system for studying targeting and assembly of both wt and mutant MPs (Bertens et al., 20...

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Plant Virus Movement

Tilsner

Taliansky

Torrance

2014

Encyclopedia of Life Sciences

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Plant virus movement is the process of the spread of virus genetic material from the initially infected cells to the rest of the plant. There are several distinct stages including intracellular movement from sites of virus replication to plasmodesmata (PD) (plant‐specific intercellular nanopores), cell‐to‐cell trafficking through PD and long‐distance movement between organs through the phloem (the specialized vascular system used by plants for the transport of assimilates and macromolecules). Transport is mediated by virus‐encoded ‘movement’ proteins. Several different types of movement proteins can be distinguished, which share properties such as nucleic acid binding, targeting and dilation of PD, and intercellular movement. These functions can be distributed amongst several movement proteins. Some movement proteins form a hollow tubule through PD which allows the passage of virus particles, but for the majority of viruses, the precise mechanism of transport through PD is still unknown. Key Concepts: Plant viruses need to overcome the barrier of the cell wall to spread from the initially infected cells to the rest of the plant. Local cell‐to‐cell movement occurs through PD which are plant‐specific membrane‐lined nanopores that connect adjoining cells. Long‐distance movement between plant organs occurs through the phloem, one of the two components of the plant's vasculature. Viral cell‐to‐cell movement constitutes an extreme population bottleneck and needs to stay ahead of plant defence signals trafficking through the same channels. Movement is mediated by one or several virus‐encoded movement proteins, which facilitate transport of virus genomes or virus particles between cells. The transported form of the viral genome is usually either a virus particle that can be modified, or a nonvirion ribonucleoprotein complex containing RNA and movement proteins. Local and long‐distance movement forms can be different. Two different movement mechanisms are observed: insertion of a MP‐comprised tubule into PD that forms a conduit for virus particles, or tubule‐less movement by an unknown mechanism. Mechanisms contributing to PD dilation may include breakdown of the polysaccharide callose in the surrounding cell wall and severing of actin filaments. Viruses use elements of the cell's cytoskeleton and/or endomembrane system to get from replication sites to PD, or replication and movement can be spatially linked at PD. Nuclear host factors are required for long‐distance movement of some viruses.

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Evidence for the Involvement of the 58K and 48K Proteins in the Intercellular Movement of Cowpea Mosaic Virus

Cited by 136 publications

References 21 publications

Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts

Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts

Studies on the origin and structure of tubules made by the movement protein of Cowpea mosaic virus

Plant Virus Movement

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