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THE PLANT CELL, Vol 5, Issue 8 973-982, Copyright © 1993 by American Society of Plant Biologists
Alanine Scanning Mutagenesis of a Plant Virus Movement Protein Identifies Three Functional Domains
D. Giesman-Cookmeyer and S. A. Lommel
Department of Plant Pathology, Box 7616, North Carolina State University, Raleigh, North Carolina 27695-7616
Alanine scanning mutagenesis was performed on the red clover necrotic
mosaic virus (RCNMV) movement protein (MP), and 12 mutants were assayed in
vitro for RNA binding characteristics and in vivo for their ability to
potentiate RCNMV cell-to-cell movement. The mutant phenotypes that were
identified in vitro and in vivo suggest both that cooperative RNA binding
is not necessary for cell-to-cell movement in vivo and that only a fraction
of the wild-type RNA binding may be required. The MP mutants defined at
least three distinct functional regions in the MP: an RNA binding domain, a
cooperative RNA binding domain, and a third domain that is necessary for
cell-to-cell movement in vivo. This third domain may be required for
targeting the MP to cell walls and plasmodesmata, interacting with host
proteins, folding, or possibly binding RNA into a functional
ribonucleoprotein complex capable of cell-to-cell movement.
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