Abstract
Tobacco mosaic virus movement protein P30 complexes with genomic viral RNA for transport through plasmodesmata, the plant intercellular connections. Although most research with P30 focuses on its targeting to and gating of plasmodesmata, the mechanisms of P30 intracellular movement to plasmodesmata have not been defined. To examine P30 intracellular localization, we used tobacco protoplasts, which lack plasmodesmata, for transfection with plasmids carrying P30 coding sequences under a constitutive promoter and for infection with tobacco mosaic virus particles. In both systems, P30 appears as filaments that colocalize primarily with microtubules. To a lesser extent, P30 filaments colocalize with actin filaments, and in vitro experiments suggested that P30 can bind directly to actin and tubulin. This association of P30 with cytoskeletal elements may play a critical role in intracellular transport of the P30-viral RNA complex through the cytoplasm to and possibly through plasmodesmata.
Full Text
The Full Text of this article is available as a PDF (4.9 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Atkinson S. J., Doberstein S. K., Pollard T. D. Moving off the beaten track. Curr Biol. 1992 Jun;2(6):326–328. doi: 10.1016/0960-9822(92)90896-i. [DOI] [PubMed] [Google Scholar]
- Bassell G. J., Taneja K. L., Kislauskis E. H., Sundell C. L., Powers C. M., Ross A., Singer R. H. Actin filaments and the spatial positioning of mRNAS. Adv Exp Med Biol. 1994;358:183–189. doi: 10.1007/978-1-4615-2578-3_17. [DOI] [PubMed] [Google Scholar]
- Blum H., Gross H. J., Beier H. The expression of the TMV-specific 30-kDa protein in tobacco protoplasts is strongly and selectively enhanced by actinomycin. Virology. 1989 Mar;169(1):51–61. doi: 10.1016/0042-6822(89)90040-8. [DOI] [PubMed] [Google Scholar]
- Chalfie M., Tu Y., Euskirchen G., Ward W. W., Prasher D. C. Green fluorescent protein as a marker for gene expression. Science. 1994 Feb 11;263(5148):802–805. doi: 10.1126/science.8303295. [DOI] [PubMed] [Google Scholar]
- Citovsky V., Knorr D., Schuster G., Zambryski P. The P30 movement protein of tobacco mosaic virus is a single-strand nucleic acid binding protein. Cell. 1990 Feb 23;60(4):637–647. doi: 10.1016/0092-8674(90)90667-4. [DOI] [PubMed] [Google Scholar]
- Citovsky V., McLean B. G., Zupan J. R., Zambryski P. Phosphorylation of tobacco mosaic virus cell-to-cell movement protein by a developmentally regulated plant cell wall-associated protein kinase. Genes Dev. 1993 May;7(5):904–910. doi: 10.1101/gad.7.5.904. [DOI] [PubMed] [Google Scholar]
- Citovsky V., Wong M. L., Shaw A. L., Prasad B. V., Zambryski P. Visualization and characterization of tobacco mosaic virus movement protein binding to single-stranded nucleic acids. Plant Cell. 1992 Apr;4(4):397–411. doi: 10.1105/tpc.4.4.397. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Citovsky V., Zambryski P. How do plant virus nucleic acids move through intercellular connections? Bioessays. 1991 Aug;13(8):373–379. doi: 10.1002/bies.950130802. [DOI] [PubMed] [Google Scholar]
- Citovsky V., Zambryski P. Transport of nucleic acids through membrane channels: snaking through small holes. Annu Rev Microbiol. 1993;47:167–197. doi: 10.1146/annurev.mi.47.100193.001123. [DOI] [PubMed] [Google Scholar]
- Citovsky V., Zupan J., Warnick D., Zambryski P. Nuclear localization of Agrobacterium VirE2 protein in plant cells. Science. 1992 Jun 26;256(5065):1802–1805. doi: 10.1126/science.1615325. [DOI] [PubMed] [Google Scholar]
- Cooper J. A. Effects of cytochalasin and phalloidin on actin. J Cell Biol. 1987 Oct;105(4):1473–1478. doi: 10.1083/jcb.105.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deom C. M., Lapidot M., Beachy R. N. Plant virus movement proteins. Cell. 1992 Apr 17;69(2):221–224. doi: 10.1016/0092-8674(92)90403-y. [DOI] [PubMed] [Google Scholar]
- Deom C. M., Oliver M. J., Beachy R. N. The 30-kilodalton gene product of tobacco mosaic virus potentiates virus movement. Science. 1987 Jul 24;237(4813):389–394. doi: 10.1126/science.237.4813.389. [DOI] [PubMed] [Google Scholar]
- Deom C. M., Schubert K. R., Wolf S., Holt C. A., Lucas W. J., Beachy R. N. Molecular characterization and biological function of the movement protein of tobacco mosaic virus in transgenic plants. Proc Natl Acad Sci U S A. 1990 May;87(9):3284–3288. doi: 10.1073/pnas.87.9.3284. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ding B., Haudenshield J. S., Hull R. J., Wolf S., Beachy R. N., Lucas W. J. Secondary plasmodesmata are specific sites of localization of the tobacco mosaic virus movement protein in transgenic tobacco plants. Plant Cell. 1992 Aug;4(8):915–928. doi: 10.1105/tpc.4.8.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fromm M., Taylor L. P., Walbot V. Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5824–5828. doi: 10.1073/pnas.82.17.5824. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hesketh J. Translation and the cytoskeleton: a mechanism for targeted protein synthesis. Mol Biol Rep. 1994 May;19(3):233–243. doi: 10.1007/BF00986965. [DOI] [PubMed] [Google Scholar]
- Howard E. A., Zupan J. R., Citovsky V., Zambryski P. C. The VirD2 protein of A. tumefaciens contains a C-terminal bipartite nuclear localization signal: implications for nuclear uptake of DNA in plant cells. Cell. 1992 Jan 10;68(1):109–118. doi: 10.1016/0092-8674(92)90210-4. [DOI] [PubMed] [Google Scholar]
- Karasev A. V., Kashina A. S., Gelfand V. I., Dolja V. V. HSP70-related 65 kDa protein of beet yellows closterovirus is a microtubule-binding protein. FEBS Lett. 1992 Jun 8;304(1):12–14. doi: 10.1016/0014-5793(92)80578-5. [DOI] [PubMed] [Google Scholar]
- Koonin E. V., Dolja V. V. Evolution and taxonomy of positive-strand RNA viruses: implications of comparative analysis of amino acid sequences. Crit Rev Biochem Mol Biol. 1993;28(5):375–430. doi: 10.3109/10409239309078440. [DOI] [PubMed] [Google Scholar]
- Langford G. M. Actin- and microtubule-dependent organelle motors: interrelationships between the two motility systems. Curr Opin Cell Biol. 1995 Feb;7(1):82–88. doi: 10.1016/0955-0674(95)80048-4. [DOI] [PubMed] [Google Scholar]
- Lehto K., Bubrick P., Dawson W. O. Time course of TMV 30K protein accumulation in intact leaves. Virology. 1990 Jan;174(1):290–293. doi: 10.1016/0042-6822(90)90077-5. [DOI] [PubMed] [Google Scholar]
- Luby-Phelps K. Physical properties of cytoplasm. Curr Opin Cell Biol. 1994 Feb;6(1):3–9. doi: 10.1016/0955-0674(94)90109-0. [DOI] [PubMed] [Google Scholar]
- McLean B. G., Waigmann E., Citovsky V., Zambryski P. Cell-to-cell movement of plant viruses. Trends Microbiol. 1993 Jun;1(3):105–109. doi: 10.1016/0966-842x(93)90116-9. [DOI] [PubMed] [Google Scholar]
- Melki R., Gaudin Y., Blondel D. Interaction between tubulin and the viral matrix protein of vesicular stomatitis virus: possible implications in the viral cytopathic effect. Virology. 1994 Jul;202(1):339–347. doi: 10.1006/viro.1994.1350. [DOI] [PubMed] [Google Scholar]
- Meshi T., Watanabe Y., Saito T., Sugimoto A., Maeda T., Okada Y. Function of the 30 kd protein of tobacco mosaic virus: involvement in cell-to-cell movement and dispensability for replication. EMBO J. 1987 Sep;6(9):2557–2563. doi: 10.1002/j.1460-2075.1987.tb02544.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morozov SYu, Dolja V. V., Atabekov J. G. Probable reassortment of genomic elements among elongated RNA-containing plant viruses. J Mol Evol. 1989 Jul;29(1):52–62. doi: 10.1007/BF02106181. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pasick J. M., Kalicharran K., Dales S. Distribution and trafficking of JHM coronavirus structural proteins and virions in primary neurons and the OBL-21 neuronal cell line. J Virol. 1994 May;68(5):2915–2928. doi: 10.1128/jvi.68.5.2915-2928.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Restrepo M. A., Freed D. D., Carrington J. C. Nuclear transport of plant potyviral proteins. Plant Cell. 1990 Oct;2(10):987–998. doi: 10.1105/tpc.2.10.987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shibaoka H., Nagai R. The plant cytoskeleton. Curr Opin Cell Biol. 1994 Feb;6(1):10–15. doi: 10.1016/0955-0674(94)90110-4. [DOI] [PubMed] [Google Scholar]
- Singer R. H. The cytoskeleton and mRNA localization. Curr Opin Cell Biol. 1992 Feb;4(1):15–19. doi: 10.1016/0955-0674(92)90053-f. [DOI] [PubMed] [Google Scholar]
- Smith D. E., Fisher P. A. Identification, developmental regulation, and response to heat shock of two antigenically related forms of a major nuclear envelope protein in Drosophila embryos: application of an improved method for affinity purification of antibodies using polypeptides immobilized on nitrocellulose blots. J Cell Biol. 1984 Jul;99(1 Pt 1):20–28. doi: 10.1083/jcb.99.1.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Staiger C. J., Lloyd C. W. The plant cytoskeleton. Curr Opin Cell Biol. 1991 Feb;3(1):33–42. doi: 10.1016/0955-0674(91)90163-s. [DOI] [PubMed] [Google Scholar]
- Topp K. S., Meade L. B., LaVail J. H. Microtubule polarity in the peripheral processes of trigeminal ganglion cells: relevance for the retrograde transport of herpes simplex virus. J Neurosci. 1994 Jan;14(1):318–325. doi: 10.1523/JNEUROSCI.14-01-00318.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vale R. D. Intracellular transport using microtubule-based motors. Annu Rev Cell Biol. 1987;3:347–378. doi: 10.1146/annurev.cb.03.110187.002023. [DOI] [PubMed] [Google Scholar]
- Waigmann E., Zambryski P. Tobacco mosaic virus movement protein-mediated protein transport between trichome cells. Plant Cell. 1995 Dec;7(12):2069–2079. doi: 10.1105/tpc.7.12.2069. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Watanabe Y., Emori Y., Ooshika I., Meshi T., Ohno T., Okada Y. Synthesis of TMV-specific RNAs and proteins at the early stage of infection in tobacco protoplasts: transient expression of the 30K protein and its mRNA. Virology. 1984 Feb;133(1):18–24. doi: 10.1016/0042-6822(84)90421-5. [DOI] [PubMed] [Google Scholar]
- Wilhelm J. E., Vale R. D. RNA on the move: the mRNA localization pathway. J Cell Biol. 1993 Oct;123(2):269–274. doi: 10.1083/jcb.123.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Williamson R. E. Organelle Movements along Actin Filaments and Microtubules. Plant Physiol. 1986 Nov;82(3):631–634. doi: 10.1104/pp.82.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wolf S., Deom C. M., Beachy R. N., Lucas W. J. Movement protein of tobacco mosaic virus modifies plasmodesmatal size exclusion limit. Science. 1989 Oct 20;246(4928):377–379. doi: 10.1126/science.246.4928.377. [DOI] [PubMed] [Google Scholar]
- Wolf S., Deom C. M., Beachy R., Lucas W. J. Plasmodesmatal function is probed using transgenic tobacco plants that express a virus movement protein. Plant Cell. 1991 Jun;3(6):593–604. doi: 10.1105/tpc.3.6.593. [DOI] [PMC free article] [PubMed] [Google Scholar]