Intracellular distribution of cowpea mosaic virus movement protein as visualised by green fluorescent protein fusions.
Cowpea mosaic virus (CPMV) derivatives expressing movement protein (MP) green fluorescent protein (GFP) fusions (MP:GFP) were used to study the intracellular targeting and localization of the MP in cowpea protoplasts and plants. In protoplasts, a virus coding for a wild type MP:GFP (MPfGFP) induced the formation of fluorescent tubular structures, which shows that subcellular targeting and tubule formation are not affected by fusion of GFP to the C-terminus of the MP. In plants, MPfGFP infections were mostly confined to single epidermal cells and failed to achieve a systemic infection, probably because the fusion of GFP to the MP interfered with MP-virion interaction. MP:GFP mainly accumulated in fluorescent spots in the cell wall of epidermal cells of inoculated leaves, which may represent short tubular structures in modified plasmodesmata. At the cuticle-side of epidermal cells tubular structures were detected indicating that tubule formation in plants, as in protoplasts, does not require the presence of functional plasmodesmata. Furthermore, results were obtained which indicate that CPMV MP:GFP is able to traffic from cell-to-cell by itself. The possible significance of this finding is discussed.
Laboratory of Molecular Biology, Wageningen University, Wageningen, The Netherlands.
K
K
Gopinath
P
P
Bertens
J
J
Pouwels
H
H
Marks
J
J
Van Lent
J
J
Wellink
A
A
Van Kammen
D017798:Comovirus / Q000737:chemistry*; D007887:Fabaceae / Q000821:virology*; D005816:Genetic Complementation Test; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000032:analysis; D018515:Plant Leaves / Q000821:virology; D053830:Plant Viral Movement Proteins; D011523:Protoplasts / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000032:analysis*; D014764:Viral Proteins / Q000032:analysis*
D008164:Luminescent Proteins; D053830:Plant Viral Movement Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins