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THE PLANT CELL, Vol 4, Issue 4 397-411, Copyright © 1992 by American Society of Plant Biologists
Visualization and Characterization of Tobacco Mosaic Virus Movement Protein Binding to Single-Stranded Nucleic Acids
V. Citovsky, M. L. Wong, A. L. Shaw, BVV. Prasad and P. Zambryski
Department of Plant Biology, University of California, Berkeley, California 94720
Cell-to-cell spread of tobacco mosaic virus (TMV) is presumed to occur
through plant intercellular connections, the plasmodesmata. Viral movement
is an active process mediated by a specific virus-encoded P30 protein. P30
has at least two functions, to cooperatively bind single-stranded nucleic
acids and to increase plasmodesmatal permeability. Here, we visualized P30
complexes with single-stranded DNA and RNA. These complexes are long,
unfolded, and very thin (1.5 to 2.0 nm in diameter). Unlike TMV virions
(300 x 18 nm), the complexes are compatible in size with the P30-induced
increase in plasmodesmatal permeability (2.4 to 3.1 nm), making them likely
candidates for the structures involved in the cell-to-cell movement of TMV.
Mutational analysis using single and double deletion mutants of P30
revealed three regions potentially important for the protein function.
Amino acid residues 65 to 86 possibly are required for correct folding of
the active protein, and the regions between amino acid residues 112 to 185
and 185 to 268 potentially contain two independently active single-stranded
nucleic acid binding domains designated binding domains A and B,
respectively.
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