A modified form of the alfalfa mosaic virus movement protein induces stressed phenotypes in transgenic tobacco

1996 ◽  
Vol 74 (6) ◽  
pp. 939-951 ◽  
Author(s):  
O. Rohfritsch ◽  
A. Poirson ◽  
M.-J. Gagey ◽  
C. Stussi-Garaud ◽  
T. Godefroy-Colburn ◽  
...  

A modified form of the movement protein (P3) of alfalfa mosaic virus, lacking amino acids 21 to 34, was transgenically expressed in Nicotiana tabacum cv. Xanthi (genotype nn) and cv. Xanthi nc (genotype NN). The modified protein (designated P3Δ[21–34]) was expressed more strongly than the full-length protein. The localization of P3Δ[21–34] was investigated by subcellular fractionation and immunocytochemistry. Immunolabelling was most frequent in vascular parenchymal cells, mainly in the cytoplasm (endoplasmic reticulum, Golgi, plasma membrane vesicles) but also in the cell wall. In contrast, full-length P3 accumulated almost exclusively in a cell-wall enriched fraction. Transgenically expressed P3Δ[21–34] increased the plasmodesmal gating capacity of epidermal cells, as did transgenically expressed P3. Thus, the plasmodesma-gating domain of P3 does not include amino acids 21 to 34. Plants expressing P3Δ[21–34] at a high level exhibited stressed phenotypes. Phenotype 1, only observed in 'Xanthi' NN lines, was characterized by stunting, small, thick, and hairy leaves, locally high starch accumulation, and occasional necrotic cells, mainly in the bundle sheath and vascular tissue. Phenotype 2, observed in both 'Xanthi' NN and nn lines, was characterized by short internodes, numerous small green leaves, sterile flowers, regular starch accumulation, and absence of necrotic cells. The stress-inducing activity of P3Δ[21–34] may be due to either its molecular conformation or its low efficiency of export toward the cell wall. Keywords: cell to cell movement, stress reaction, plasmodesmata, transgenic plant, ultrastructure, immunocytochemistry.

2001 ◽  
Vol 14 (9) ◽  
pp. 1051-1062 ◽  
Author(s):  
Jesús A. Sánchez-Navarro ◽  
John F. Bol

The movement protein (MP) and coat protein (CP) encoded by Alfalfa mosaic virus (AMV) RNA 3 are both required for virus transport. RNA 3 vectors that expressed nonfused green fluorescent protein (GFP), MP:GPF fusions, or GFP:CP fusions were used to study the functioning of mutant MP and CP in protoplasts and plants. C-terminal deletions of up to 21 amino acids did not interfere with the function of the CP in cell-to-cell movement, although some of these mutations interfered with virion assembly. Deletion of the N-terminal 11 or C-terminal 45 amino acids did not interfere with the ability of MP to assemble into tubular structures on the protoplast surface. Additionally, N- or C-terminal deletions disrupted tubule formation. A GFP:CP fusion was targeted specifically into tubules consisting of a wild-type MP. All MP deletion mutants that showed cell-to-cell and systemic movement in plants were able to form tubular structures on the surface of protoplasts. Brome mosaic virus (BMV) MP did not support AMV transport. When the C-terminal 48 amino acids were replaced by the C-terminal 44 amino acids of the AMV MP, however, the BMV/AMV chimeric protein permitted wild-type levels of AMV transport. Apparently, the C terminus of the AMV MP, although dispensable for cell-to-cell movement, confers specificity to the transport process.


1999 ◽  
Vol 354 (1383) ◽  
pp. 637-643 ◽  
Author(s):  
Vitaly Citovsky

Cell–to–cell movement of tobacco mosaic virus (TMV) is used to illustrate macromolecular traffic through plant intercellular connections, the plasmodesmata. This transport process is mediated by a specialized viral movement protein, P30. In the initially infected cell, P30 is produced by transcription of a subgenomic RNA derived from the invading virus. Presumably, P30 then associates with a certain proportion of the viral RNA molecules, sequestering them from replication and mediating their transport into neighbouring uninfected host cells. This nucleoprotein complex is targeted to plasmodesmata, possibly via interaction with the host cell cytoskeleton. Prior to passage through a plasmodesma, the plasmodesmal channel is dilated by the movement protein. It is proposed that targeting of P30–TMV RNA complexes to plasmodesmata involves binding to a specific cell wall–associated receptor molecule. In addition, a cell wall–associated protein kinase, phosphorylates P30 at its carboxy–terminus and minimizes P30–induced interference with plasmodesmatal permeability during viral infection.


Virology ◽  
2005 ◽  
Vol 333 (1) ◽  
pp. 10-21 ◽  
Author(s):  
Douglas Tremblay ◽  
Andrew A. Vaewhongs ◽  
Katherine A. Turner ◽  
Tim L. Sit ◽  
Steven A. Lommel

2001 ◽  
Vol 75 (17) ◽  
pp. 8045-8053 ◽  
Author(s):  
Hideaki Nagano ◽  
Kazuyuki Mise ◽  
Iwao Furusawa ◽  
Tetsuro Okuno

ABSTRACT Plant viruses have movement protein (MP) gene(s) essential for cell-to-cell movement in hosts. Cucumber mosaic virus (CMV) requires its own coat protein (CP) in addition to the MP for intercellular movement. Our present results using variants of both CMV and a chimeric Brome mosaic virus with the CMV MP gene revealed that CMV MP truncated in its C-terminal 33 amino acids has the ability to mediate viral movement independently of CP. Coexpression of the intact and truncated CMV MPs extremely reduced movement of the chimeric viruses, suggesting that these heterogeneous CMV MPs function antagonistically. Sequential deletion analyses of the CMV MP revealed that the dispensability of CP occurred when the C-terminal deletion ranged between 31 and 36 amino acids and that shorter deletion impaired the ability of the MP to promote viral movement. This is the first report that a region of MP determines the requirement of CP in cell-to-cell movement of a plant virus.


2013 ◽  
Vol 94 (3) ◽  
pp. 677-681 ◽  
Author(s):  
Thor V. M. Fajardo ◽  
Ana Peiró ◽  
Vicente Pallás ◽  
Jesús Sánchez-Navarro

We previously showed that the movement protein (MP) gene of Alfalfa mosaic virus (AMV) is functionally exchangeable for the cell-to-cell transport of the corresponding genes of Tobacco mosaic virus (TMV), Brome mosaic virus, Prunus necrotic ringspot virus, Cucumber mosaic virus and Cowpea mosaic virus. We have analysed the capacity of the heterologous MPs to systemically transport the corresponding chimeric AMV genome. All MPs were competent in systemic transport but required the fusion at their C terminus of the coat protein-interacting C-terminal 44 aa (A44) of the AMV MP. Except for the TMV MP, the presence of the hybrid virus in upper leaves correlated with the capacity to move locally. These results suggest that all the MPs assigned to the 30K superfamily should be exchangeable not only for local virus movement but also for systemic transport when the A44 fragment is present.


Virology ◽  
1992 ◽  
Vol 188 (2) ◽  
pp. 896-899 ◽  
Author(s):  
Fabrice Schoumacher ◽  
Claude Erny ◽  
Anne Berna ◽  
Therese Godefroy-Colburn ◽  
Christiane Stussi-Garaud

2003 ◽  
Vol 77 (20) ◽  
pp. 10790-10798 ◽  
Author(s):  
A. Corina Vlot ◽  
Sebastiaan M. Laros ◽  
John F. Bol

ABSTRACT RNAs 1 and 2 of the tripartite genome of alfalfa mosaic virus encode the replicase proteins P1 and P2, respectively, whereas RNA 3 encodes the movement protein and coat protein. Transient expression of wild-type (wt) and mutant viral RNAs and proteins by agroinfiltration of plant leaves was used to study cis- and trans-acting functions of the helicase-like domain in P1 and the polymerase-like domain in P2. Three mutations in conserved motifs of the helicase-like domain of P1 affected one or more steps leading to synthesis of minus-strand RNAs 1, 2, and 3. In leaves containing transiently expressed P1 and P2, replication of wt but not mutant RNA 1 was observed. Apparently, the transiently expressed P1 could not complement the defect in replication of the RNA 1 mutant. Moreover, the transiently expressed wt replicase supported replication of RNA 2, but this replication was blocked in trans by coexpression of mutant RNA 1. However, expression of mutant RNA 1 did not interfere with the replication of RNA 3 by the wt replicase. Similarly, a mutation in the GDD motif encoded by RNA 2 could not be complemented in trans and affected the replication of RNA 1 by a wt replicase, while replication of RNA 3 remained unaffected. In competition assays, the transient wt replicase preferentially replicated RNA 3 over RNAs 1 and 2. The results indicate that one or more functions of P1 and P2 act in cis and point to the existence of a mechanism that coordinates the replication of RNAs 1 and 2.


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