Role of Microtubules in the Intracellular Distribution of Tobacco Mosaic Virus Movement Protein

Despite its central role in virus infection, little is known about the mechanisms of intracellular trafficking of virus components within infected cells. In this study, we followed the dynamics of tobacco mosaic virus movement protein (MP) distribution in living protoplasts after disruption of micro...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2000-10, Vol.97 (22), p.12345-12349
Main Authors: Mas, Paloma, Beachy, Roger N.
Format: Article
Language:English
Subjects:
Antibodies
Biological Sciences
Cells
Cytoskeleton
Fluorescence
Fluorescent Antibody Technique
Green Fluorescent Proteins
In Situ Hybridization
Infections
Luminescent Proteins - genetics
Microscopy, Confocal
Microtubules - physiology
movement protein
Mutation
Nicotiana - ultrastructure
Nicotiana - virology
Plant cells
Plant Viral Movement Proteins
Plants, Toxic
Proteins
Protoplasts
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
RNA
RNA, Viral - metabolism
Tobacco
Tobacco mosaic virus
Viral Proteins - genetics
Viral Proteins - metabolism
Virus replication
Viruses
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Summary:Despite its central role in virus infection, little is known about the mechanisms of intracellular trafficking of virus components within infected cells. In this study, we followed the dynamics of tobacco mosaic virus movement protein (MP) distribution in living protoplasts after disruption of microtubules (MTs) by cold treatment and subsequent rewarming to 29 degrees C. At early stages of infection, cold treatment (4 degrees C) caused the accumulation of MP fused to green fluorescent protein (GFP) in large virus replication bodies that localized in perinuclear positions, whereas at midstages of infection, the association of MP:GFP with MTs was disrupted. Rewarming the protoplasts to 29 degrees C reestablished the association of MTs with the replication bodies that subsequently spread throughout the cytoplasm and to the periphery of the cell. The role of MTs in the intracellular distribution of the MP also was analyzed by examining the distribution pattern of a nonfunctional mutant of MP (TAD5). Like MP:GFP, TAD5:GFP interacted with the endoplasmic reticulum membranes and colocalized with its viral RNA but did not colocalize with MTs. The involvement of MTs in the intracellular distribution of tobacco mosaic virus MP is discussed.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.97.22.12345