Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2002 Apr;76(8):3974-80.
doi: 10.1128/jvi.76.8.3974-3980.2002.

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, Elena Suslova, Jacqueline Ferralli, Oliver Sterthaus, Carl M Deom, Manfred Heinlein
Affiliations

Intramolecular complementing mutations in tobacco mosaic virus movement protein confirm a role for microtubule association in viral RNA transport

Vitaly Boyko et al. J Virol. 2002 Apr.

Abstract

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.

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
Local lesion assay on leaves of N. tabacum cv. Xanthi NN to test for infectivity and cell-to-cell movement functions of TMV-MPP81S-GFP and TMV-MPP81S;T104I;R167K-GFP. (a) TMV-MPP81S-GFP does not give rise to local lesions. (b) TMV-MPP81S-GFP produces local lesions on plants that are transgenic for MP. (c) TMV-MPP81S;T104I;R167K-GFP triggers the formation of small local lesions on wild-type plants.
FIG. 2.
FIG. 2.
Patterns of subcellular distribution of MPP81S-GFP and MPP81S;T104I;R167K-GFP observed in infected protoplasts at 20 hpi. (A) TMV-MPP81S-GFP-infected protoplast. MPP81S-GFP does not localize to any discernible intracellular structure. (B) TMV-MPP81S;T104I;R167K-GFP-infected protoplasts. MPP81S;T104I;R167K-GFP localizes to inclusion bodies (a), microtubules (b), peripheral puncta (c), and hair-like protrusions (d). Bar, 10 μm.
FIG. 3.
FIG. 3.
Subcellular distribution of MPP81S-GFP and MPP81S;T104I;R167K-GFP in leaf epidermal cells of N. benthamiana. (a) Infection site of TMV-MPP81S-GFP on MP-transgenic plant. (b and c) MPP81S-GFP accumulates only in Pd. (d) Infection site of TMV-MPP81S;T104I;R167K-GFP. (e) MPP81S;T104I;R167K-GFP in association with inclusion bodies and Pd (arrow). (f) MPP81S;T104I;R167K-GFP associated with microtubules and Pd (arrows). Bars, 1 mm (a and d) and 10 μm (b, c, e, and f).
FIG. 4.
FIG. 4.
RNA-binding properties of wild-type and mutant MPs purified from E. coli. 32P-labeled RNA (10 nM; see Materials and Methods) was incubated with increasing amounts of MP:His6wt (a), MP:His6P81S (b), and MP:His6P81S;T104I;R167K (c) as indicated. Resulting protein-RNA complexes were visualized by nondenaturing electrophoresis and autoradiography.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Aaziz, R., S. Dinant, and B. L. Epel. 2001. Plasmodesmata and plant cytoskeleton. Trends Plant Sci. 6:326-330. - PubMed
    1. Atkins, D., R. Hull, B. Wells, K. Roberts, P. Moore, and R. N. Beachy. 1991. The tobacco mosaic virus 30K movement protein in transgenic tobacco plants is localized to plasmodesmata. J. Gen. Virol. 72:209-211. - PubMed
    1. Bassell, G., and R. H. Singer. 1997. mRNA and cytoskeletal elements. Curr. Opin. Cell Biol. 9:109-115. - PubMed
    1. Beachy, R. N., and M. Heinlein. 2000. Role of P30 in replication and spread of TMV. Traffic 1:540-544. - PubMed
    1. Berna, A., R. Gafny, S. Wolf, W. J. Lucas, C. A. Holt, and R. N. Beachy. 1991. The TMV movement protein: role of the C-terminal 73 amino acids in subcellular localization and function. Virology 182:682-689. - PubMed

MeSH terms

LinkOut - more resources