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Journal of Virology, April 2002, p. 3974-3980, Vol. 76, No. 8
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.8.3974-3980.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Intramolecular Complementing Mutations in Tobacco Mosaic Virus Movement Protein Confirm a Role for Microtubule Association in Viral RNA Transport

Vitaly Boyko,1 Jamie Alan Ashby,1 Elena Suslova,1 Jacqueline Ferralli,1 Oliver Sterthaus,1 Carl M. Deom,2 and Manfred Heinlein1*

Friedrich Miescher Institute for Biomedical Research, Novartis Research Foundation, CH-4058 Basel, Switzerland,1 Department of Plant Pathology, The University of Georgia, Athens, Georgia 306022

Received 2 November 2001/ Accepted 22 January 2002

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 MPP81S-GFP and MPP81S;T104I;R167K-GFP differ in subcellular distribution. MPP81S-GFP lacks specific sites of accumulation in protoplasts and, in epidermal cells, exclusively localizes to Pd. MPP81S;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.

* Corresponding author. Mailing address: Friedrich Miescher Institute for Biomedical Research, Novartis Research Foundation, Maulbeerstrasse 66, CH-4058 Basel, Switzerland. Phone: 41 61 697 85 17. Fax: 41 61 697 39 76. E-mail: heinlein{at}fmi.ch.

Journal of Virology, April 2002, p. 3974-3980, Vol. 76, No. 8
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.8.3974-3980.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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