Domains of the TMV movement protein involved in subcellular localization

• Published in: The Plant journal : for cell and molecular biology Vol. 15; no. 1; pp. 15 - 25
• Main Authors: Kahn, Theodore W., Lapidot, Moshe, Heinlein, Manfred, Reichel, Christoph, Cooper, Bret, Gafny, Ron, Beachy, Roger N.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science, Ltd 01.07.1998
• Subjects: Amino Acid Sequence; Biological Transport; Cytoplasm - virology; Genetic Complementation Test; Green Fluorescent Proteins; Luminescent Proteins - genetics; Microscopy, Fluorescence; Microtubules - virology; Molecular Sequence Data; Nicotiana - virology; Nitrous Acid - pharmacology; Plant Viral Movement Proteins; Plants, Genetically Modified; Plants, Toxic; Protein Structure, Tertiary; Protoplasts; Recombinant Fusion Proteins; RNA, Viral - drug effects; Sequence Deletion; Tobacco Mosaic Virus - genetics; Tobacco Mosaic Virus - metabolism; Viral Proteins - chemistry; Viral Proteins - genetics; Viral Proteins - metabolism
• ISSN: 0960-7412; 1365-313X
• DOI: 10.1046/j.1365-313X.1998.00172.x

Summary To identify and map functionally important regions of the tobacco mosaic virus movement protein, deletions of three amino acids were introduced at intervals of 10 amino acids throughout the protein. Mutations located between amino acids 1 and 160 abolished the capacity of the protein to transport virus from cell to cell, while some of the mutations in the C‐terminal third of the protein permitted function. Despite extensive tests, no examples were found of intermolecular complementation between mutants, suggesting that function requires each movement protein molecule to be fully competent. Many of the mutants were fused to green fluorescent protein, and their subcellular localizations were determined by fluorescence microscopy in infected plants and protoplasts. Most mutants lost the ability to accumulate in one or more of the multiple subcellular sites targeted by wild‐type movement protein, suggesting that specific functional domains were disrupted. The order in which accumulation at subcellular sites occurs during infection does not represent a targeting pathway. Association of the movement protein with microtubules or with plasmodesmata can occur in the absence of other associations. The region of the protein around amino acids 9–11 may be involved in targeting the protein to cortical bodies (probably associated with the endoplasmic reticulum) and to plasmodesmata. The region around residues 49–51 may be involved in co‐alignment of the protein with microtubules. The region around residues 88–101 appears to play a role in targeting to both the cortical bodies and microtubules. Thus, the movement protein contains independently functional domains.