Intramolecular Complementing Mutations in Tobacco Mosaic Virus Movement Protein Confirm a Role for Microtubule Association in Viral RNA Transport
| Autor: | Jacqueline Ferralli, Jamie Ashby, Manfred Heinlein, Elena Suslova, Carl M. Deom, Vitaly Boyko, Oliver Sterthaus |
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| Jazyk: | angličtina |
| Rok vydání: | 2002 |
| Předmět: |
Recombinant Fusion Proteins
Immunology Green Fluorescent Proteins Nicotiana benthamiana Plasmodesma Biology medicine.disease_cause Microbiology Microtubules Inclusion bodies Viral Proteins Virology Tobacco medicine Tobacco mosaic virus Movement protein Mutation Protoplasts fungi Genetic Complementation Test RNA Biological Transport biology.organism_classification Molecular biology Virus-Cell Interactions Complementation Plant Leaves Plant Viral Movement Proteins Tobacco Mosaic Virus Luminescent Proteins Insect Science RNA Viral Subcellular Fractions |
| Popis: | The movement protein (MP) of Tobacco mosaic virus (TMV) facilitates the cell-to-cell transport of the viral RNA genome through plasmodesmata (Pd). A previous report described the functional reversion of a dysfunctional mutation in MP (Pro81Ser) by two additional amino acid substitution mutations (Thr104Ile and Arg167Lys). To further explore the mechanism underlying this intramolecular complementation event, the mutations were introduced into a virus derivative expressing the MP as a fusion to green fluorescent protein (GFP). Microscopic analysis of infected protoplasts and of infection sites in leaves of MP-transgenic Nicotiana benthamiana indicates that MP P81S -GFP and MP P81S;T104I;R167K -GFP differ in subcellular distribution. MP P81S -GFP lacks specific sites of accumulation in protoplasts and, in epidermal cells, exclusively localizes to Pd. MP P81S;T104I;R167K -GFP, in contrast, in addition localizes to inclusion bodies and microtubules and thus exhibits a subcellular localization pattern that is similar, if not identical, to the pattern reported for wild-type MP-GFP. Since accumulation of MP to inclusion bodies is not required for function, these observations confirm a role for microtubules in TMV RNA cell-to-cell transport. |
| Databáze: | OpenAIRE |
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