Alanine scanning mutagenesis of a plant virus movement protein identifies three functional domains.
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.
Giesman-Cookmeyer
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Department of Plant Pathology, North Carolina State University, Raleigh 27695-7616.
Lommel
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SA
D000595:Amino Acid Sequence; D002465:Cell Movement / Q000235:genetics; D005814:Genes, Viral; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D016297:Mutagenesis, Site-Directed; D053830:Plant Viral Movement Proteins; D017434:Protein Structure, Tertiary; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D053830:Plant Viral Movement Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D014764:Viral Proteins