The involvement of cowpea mosaic virus M RNA-encoded proteins in tubule formation

Kasteel, D.; Wellink, J.; Verver, J.; Lent, J.W.M. van; Goldbach, Rob; Kammen, A. van

doi:10.1099/0022-1317-74-8-1721

Cited by 57 publications

(35 citation statements)

References 13 publications

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“…; 4 h after embedding), MP-GFP had clearly accumulated in peripheral punctate spots of various sizes and in tubules, as observed previously (Gopinath et al, 2003). Since CPMV RNA2 is translated into two overlapping polyproteins due to the presence of two in-frame start codons (Pouwels et al, 2002a), GFP is also fused to the cofactor for RNA2 replication, which causes fluorescence to accumulate in the nucleus as well (Gopinath et al, 2000;Kasteel et al, 1993;Pouwels et al, 2002b). During the course of the experiment, at the site of some, mostly small, peripheral punctate spots, a tubule was formed ( Fig.…”

Section: Resultsmentioning

confidence: 67%

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: Resultsmentioning

confidence: 67%

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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“…Another mechanism involves the movement of viral genomes in the form of complete virions along the newly formed tubular structures, as shown for comoviruses, caulimoviruses, and nepoviruses (16)(17)(18)(19)(20)(21)(22).…”

Section: Discussionmentioning

confidence: 99%

“…A model proposed for TSWV movement predicts a tubule-guided mechanism to transport nucleocapsids, infectious subviral particles of viral genomic RNA complexed with nucleocapsid protein (N), and the viral polymerase (15). Involvement of tubular structures has been described for como-, nepo-, and caulimoviruses (16)(17)(18)(19)(20)(21)(22). The NSm protein shows some typical characteristics of viral movement proteins such as expression only at early stages of infection, intracellular localization of NSm close to plasmodesmata and nucleocapsid associations in the cytoplasm, and a tubule-forming capacity (15,23).…”

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confidence: 99%

The movement protein NSm of tomato spotted wilt tospovirus (TSWV): RNA binding, interaction with the TSWV N protein, and identification of interacting plant proteins

Soellick¹,

Uhrig²,

Bucher³

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

179

129

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The nonstructural NSm protein of tomato spotted wilt tospovirus (TSWV) represents a putative viral movement protein involved in cell-to-cell movement of nonenveloped ribonucleocapsid structures. To study the molecular basis of NSm function, we expressed the protein in Escherichia coli and investigated protein-protein and protein-RNA interactions of NSm protein in vitro. NSm specifically interacts with TSWV N protein and binds single-stranded RNA in a sequence-nonspecific manner. Using NSm as a bait in a yeast two-hybrid screen, we identified two homologous NSm-binding proteins of the DnaJ family from Nicotiana tabacum and Arabidopsis thaliana.

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“…Cell-to-cell movement of CPMV is characterized by transport of mature virions through tubules that are assembled inside the plasmodesmal pore and which contain the RNA-2-coded 48 kDa movement protein (MP) (Wellink & Van Kammen, 1989;Kasteel et al, 1993;Van Lent et al, 1990). Such a tubule-guided cell-to-cell transport system has been described for viruses of different genera including Caulimovirus, Nepovirus, Bromovirus and Tospovirus (Perbal et al, 1993;Wieczorek & Sanfacon, 1993;Kasteel et al, 1997;Storms et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

The C-terminal region of the movement protein of Cowpea mosaic virus is involved in binding to the large but not to the small coat protein

Carvalho

Wellink²,

Ribeiro

et al. 2003

Journal of General Virology

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Cowpea mosaic virus (CPMV) moves from cell to cell as virus particles which are translocated through a plasmodesmata-penetrating transport tubule made up of viral movement protein (MP) copies. To gain further insight into the roles of the viral MP and capsid proteins (CP) in virus movement, full-length and truncated forms of the MP were expressed in insect cells using the baculovirus expression system. Using ELISA and blot overlay assays, affinity purified MP was shown to bind specifically to intact CPMV virions and to the large CP, but not to the small CP. This binding was not observed with a C-terminal deletion mutant of the MP, although this mutant retained the capacity to bind to other MP molecules and to form tubules. These results suggest that the C-terminal 48 amino acids constitute the virion-binding domain of the MP.

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The involvement of cowpea mosaic virus M RNA-encoded proteins in tubule formation

Cited by 57 publications

References 13 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

The movement protein NSm of tomato spotted wilt tospovirus (TSWV): RNA binding, interaction with the TSWV N protein, and identification of interacting plant proteins

The C-terminal region of the movement protein of Cowpea mosaic virus is involved in binding to the large but not to the small coat protein

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