Identification of distinct steps during tubule formation by the movement protein of Cowpea mosaic virus

Pouwels, Jeroen; Kornet, Noortje; Bers, Nikkie van; Guighelaar, Teun; Lent, J. van; Bisseling, Ton; Wellink, J.

doi:10.1099/vir.0.19553-0

Cited by 30 publications

(22 citation statements)

References 27 publications

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Section: Introductionmentioning

confidence: 52%

“…In protoplasts, CPMV MP is transported to the plasma membrane where it quickly accumulates in peripheral punctate spots and tubules (Gopinath et al, 2003;Pouwels et al, 2002bPouwels et al, , 2003. Using TLM and FRAP, we have shown that peripheral punctate spots are dynamic structures, which constantly exchange MP-GFP subunits.…”

Section: Discussionmentioning

confidence: 99%

“…Since tubules in plant tissue and on protoplasts are morphologically very similar (van Lent et al, 1991) and the behaviour of mutant MPs is comparable in plant tissue and protoplasts (Gopinath et al, 2003;Pouwels et al, 2003), it is highly unlikely that tubules assemble differently in protoplasts and infected leaf cells. However, we cannot rule out the possibility that the interaction of the tubule with the surrounding plasma membrane (through a host protein) is stronger in infected plant tissue.…”

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…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, 2000Gopinath et al, 2003;Kasteel et al, 1997; Pouwels et al, 2002bPouwels et al, , 2003.Recently, a CPMV variant encoding a fusion between MP and the N terminus of the green fluorescent protein (MP-GFP) was made, which, similar to non-fused MP, accumulated in the cell wall of infected leaf tissue and formed tubules on protoplasts (Gopinath et al, 2003; Pouwels et al, 2002b). Electron microscopy analysis revealed that these tubules were morphologically indistinguishable from tubules made by non-fused MP, except that GFP, which was fused to the C terminus of MP and thus present inside the tubule (Carvalho et al, 2003), prevented the incorporation of virus particles (Gopinath et al, 2003).…”

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confidence: 99%

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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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Section: Introductionmentioning

confidence: 52%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

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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Morphological Characterization of the Self‐Assembly of Virus Movement Proteins into Nanotubes in the Absence of Virus Particles

et al. 2017

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One infection mechanism of plant viruses is the generation of nanotubes by viral movement proteins, allowing cell‐to‐cell virus particle transport. Previously, it was assumed that viral nanotubes extend directly from the host‐cell plasma membrane. In virus‐infected plants, these nanotubes reach an extraordinary diameter:length ratio (≈100 nm:µm or mm range). Here, viral nanotubes are produced in a transient protoplast system; the coding sequence for alfalfa mosaic virus movement protein is translationally fused to green fluorescent protein. The maximum extension of viral nanotubes into the culture medium is achieved 24–48 h posttransfection, with lengths in the micro‐ and millimeter ranges. Scanning electron microscopy and transmission electron microscopy show that strong inhomogeneous viral nanotubes are formed compared to particle‐filled systems. The nanotubes have similar length, but fluctuating wall thickness and diameter and are susceptible to entanglement and recombination. Indirect methods demonstrate that movement proteins assemble independently at the top of the nanotube. These viral nanotubes grow distinctly from previously known natural particle‐filled systems and are a unique biological tubular nanomaterial that has the potential for micro‐ or nanoapplications as a mechanically stable structural component.

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Tubule-Guided Movement of Plant Viruses

Ritzenthaler

Hofmann

Viral Transport in Plants

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Identification of distinct steps during tubule formation by the movement protein of Cowpea mosaic virus

Cited by 30 publications

References 27 publications

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

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

Morphological Characterization of the Self‐Assembly of Virus Movement Proteins into Nanotubes in the Absence of Virus Particles

Tubule-Guided Movement of Plant Viruses

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