P3N-PIPO Interacts with P3 via the Shared N-Terminal Domain To Recruit Viral Replication Vesicles for Cell-to-Cell Movement

ABSTRACT P3N-PIPO, the only dedicated movement protein (MP) of potyviruses, directs cylindrical inclusion (CI) protein from the cytoplasm to the plasmodesma (PD), where CI forms conical structures for intercellular movement. To better understand potyviral cell-to-cell movement, we further characterized P3N-PIPO using Turnip mosaic virus (TuMV) as a model virus. We found that P3N-PIPO interacts with P3 via the shared P3N domain and that TuMV mutants lacking the P3N domain of either P3N-PIPO or P3 are defective in cell-to-cell movement. Moreover, we found that the PIPO domain of P3N-PIPO is sufficient to direct CI to the PD, whereas the P3N domain is necessary for localization of P3N-PIPO to 6K2-labeled vesicles or aggregates. Finally, we discovered that the interaction between P3 and P3N-PIPO is essential for the recruitment of CI to cytoplasmic 6K2-containing structures and the association of 6K2-containing structures with PD-located CI inclusions. These data suggest that both P3N and PIPO domains are indispensable for potyviral cell-to-cell movement and that the 6K2 vesicles in proximity to PDs resulting from multipartite interactions among 6K2, P3, P3N-PIPO, and CI may also play an essential role in this process. IMPORTANCE Potyviruses include numerous economically important viruses that represent approximately 30% of known plant viruses. However, there is still limited information about the mechanism of potyviral cell-to-cell movement. Here, we show that P3N-PIPO interacts with and recruits CI to the PD via the PIPO domain and interacts with P3 via the shared P3N domain. We further report that the interaction of P3N-PIPO and P3 is associated with 6K2 vesicles and brings the 6K2 vesicles into proximity with PD-located CI structures. These results support the notion that the replication and cell-to-cell movement of potyviruses are processes coupled by anchoring viral replication complexes at the entrance of PDs, which greatly increase our knowledge of the intercellular movement of potyviruses.

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Conversion in the Requirement of Coat Protein in Cell-to-Cell Movement Mediated by the Cucumber Mosaic Virus Movement Protein

Journal of Virology ◽

10.1128/jvi.75.17.8045-8053.2001 ◽

2001 ◽

Vol 75 (17) ◽

pp. 8045-8053 ◽

Cited By ~ 47

Author(s):

Hideaki Nagano ◽

Kazuyuki Mise ◽

Iwao Furusawa ◽

Tetsuro Okuno

Keyword(s):

Amino Acids ◽

Coat Protein ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Movement Protein ◽

Cell Movement ◽

Plant Viruses ◽

Brome Mosaic Virus ◽

Viral Movement ◽

Intercellular Movement

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.

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Actin Cytoskeleton Is Involved in Targeting of a Viral Hsp70 Homolog to the Cell Periphery

Journal of Virology ◽

10.1128/jvi.79.22.14421-14428.2005 ◽

2005 ◽

Vol 79 (22) ◽

pp. 14421-14428 ◽

Cited By ~ 62

Author(s):

Alexey I. Prokhnevsky ◽

Valera V. Peremyslov ◽

Valerian V. Dolja

Keyword(s):

Actin Cytoskeleton ◽

Tobacco Mosaic Virus ◽

Mosaic Virus ◽

Movement Protein ◽

Fluorescent Proteins ◽

Cell Movement ◽

Plant Viruses ◽

Cell Periphery ◽

Virus Particles ◽

Shock Proteins

ABSTRACT The cell-to-cell movement of plant viruses involves translocation of virus particles or nucleoproteins to and through the plasmodesmata (PDs). As we have shown previously, the movement of the Beet yellows virus requires the concerted action of five viral proteins including a homolog of cellular ∼70-kDa heat shock proteins (Hsp70h). Hsp70h is an integral component of the virus particles and is also found in PDs of the infected cells. Here we investigate subcellular distribution of Hsp70h using transient expression of Hsp70h fused to three spectrally distinct fluorescent proteins. We found that fluorophore-tagged Hsp70h forms motile granules that are associated with actin microfilaments, but not with microtubules. In addition, immobile granules were observed at the cell periphery. A pairwise appearance of these granules at the opposite sides of cell walls and their colocalization with the movement protein of Tobacco mosaic virus indicated an association of Hsp70h with PDs. Treatment with various cytoskeleton-specific drugs revealed that the intact actomyosin motility system is required for trafficking of Hsp70h in cytosol and its targeting to PDs. In contrast, none of the drugs interfered with the PD localization of Tobacco mosaic virus movement protein. Collectively, these findings suggest that Hsp70h is translocated and anchored to PDs in association with the actin cytoskeleton.

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Stability in vitro of the 69K movement protein of Turnip yellow mosaic virus is regulated by the ubiquitin-mediated proteasome pathway

Journal of General Virology ◽

10.1099/0022-1317-83-12-3187 ◽

2002 ◽

Vol 83 (12) ◽

pp. 3187-3197 ◽

Cited By ~ 37

Author(s):

Gabrièle Drugeon ◽

Isabelle Jupin

Keyword(s):

Mosaic Virus ◽

Movement Protein ◽

Cell Movement ◽

Plant Viruses ◽

Yellow Mosaic Virus ◽

Turnip Yellow Mosaic Virus ◽

Movement Proteins ◽

Transient Nature ◽

Reticulocyte Lysate

Plant viruses move to adjacent cells with the use of virus-encoded cell-to-cell movement proteins. Using proteins produced by in vitro translation, we present evidence that the ‘69K’ movement protein of Turnip yellow mosaic virus (TYMV) is recognized as a substrate for the attachment of polyubiquitin chains and for subsequent rapid and selective proteolysis by the proteasome, the ATP-dependent proteolytic system present in reticulocyte lysate. Truncation of the 69K protein suggests the existence of two degradation signals within its sequence. We propose that selective degradation of virus movement proteins may contribute to the previously reported transient nature of their accumulation during infection.

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Cylindrical Inclusion Protein of Turnip Mosaic Virus Serves as a Docking Point for the Intercellular Movement of Viral Replication Vesicles

PLANT PHYSIOLOGY ◽

10.1104/pp.17.01484 ◽

2017 ◽

Vol 175 (4) ◽

pp. 1732-1744 ◽

Cited By ~ 17

Author(s):

Nooshin Movahed ◽

Camilo Patarroyo ◽

Jiaqi Sun ◽

Hojatollah Vali ◽

Jean-François Laliberté ◽

...

Keyword(s):

Viral Replication ◽

Mosaic Virus ◽

Turnip Mosaic Virus ◽

Cylindrical Inclusion ◽

Inclusion Protein ◽

Intercellular Movement ◽

Cylindrical Inclusion Protein

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Faculty Opinions recommendation of Remorin interacting with PCaP1 impairs Turnip mosaic virus intercellular movement but is antagonised by VPg.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.736814198.793571748 ◽

2020 ◽

Author(s):

Christiaan van der Schoot

Keyword(s):

Mosaic Virus ◽

Turnip Mosaic Virus ◽

Intercellular Movement

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Tobacco mosaic virus: a pioneer to cell–to–cell movement

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.1999.0415 ◽

1999 ◽

Vol 354 (1383) ◽

pp. 637-643 ◽

Cited By ~ 39

Author(s):

Vitaly Citovsky

Keyword(s):

Cell Wall ◽

Tobacco Mosaic Virus ◽

Mosaic Virus ◽

Movement Protein ◽

Cell Movement ◽

Host Cells ◽

Specific Cell ◽

Viral Movement Protein ◽

Rna Molecules ◽

Rna Complexes

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.

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