Intracellular distribution, cell-to-cell trafficking and tubule-inducing activity of the 50 kDa movement protein of fused to green fluorescent protein Free

Abstract

The 50 kDa protein (50KP) encoded by ORF2 of (ACLSV) fused to green fluorescent protein (GFP) was expressed transiently in cells of and leaves. Its intracellular distribution, cell-to-cell trafficking in leaf epidermis and tubule formation on the surface of protoplasts were analysed. The 50KP–GFP fluorescence was distributed as small irregular spots or a fibrous network structure on the periphery of epidermal cells and protoplasts of both plant species. In leaf epidermis of , the protein spread from the cells that produced it into neighbouring cells in both young and mature leaves and targetted plasmodesmata in these cells. In contrast, GFP was restricted to single cells in most cases in mature leaves. When 50KP and GFP were co-expressed in leaf epidermis of , GFP spread more widely from the initial cells that produced it than when GFP was expressed alone, suggesting that 50KP facilitated the cell-to-cell trafficking of GFP. 50KP–GFP was able to complement local spread of 50KP-deficient virus when expressed transiently in leaf epidermis of . Expression of 50KP–GFP in protoplasts resulted in the production of tubular structures protruding from the surface. Mutational analyses showed that the C-terminal region (aa 287–457) was not essential for localization to plasmodesmata, cell-to-cell trafficking, complementation of movement of 50KP-deficient virus or tubule formation on protoplasts. In contrast, deletions in the N-terminal region resulted in the complete disruption of all these activities.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-81-8-2085
2000-08-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/81/8/0812085a.html?itemId=/content/journal/jgv/10.1099/0022-1317-81-8-2085&mimeType=html&fmt=ahah

References

  1. Atkins D., Hull R., Wells B., Roberts K., Moore P., Beachy R. N. 1991; The tobacco mosaic virus 30K movement protein in transgenic tobacco plants is localized to plasmodesmata. Journal of General Virology 72:209–211
    [Google Scholar]
  2. Canto T., Palukaitis P. 1999; Are tubules generated by the 3a protein necessary for cucumber mosaic virus movement?Molecular Plant–Microbe Interactions. 12985–993
  3. Carrington J. C., Kasschau K. D., Mahajan S. K., Schaad M. C. 1996; Cell-to-cell and long-distance transport of viruses in plants. Plant Cell 8:1669–1681
    [Google Scholar]
  4. Citovsky V., Knorr D., Schuster G., Zambryski P. 1990; The P30 movement protein of tobacco mosaic virus is a single-strand nucleic acid binding protein. Cell 60:637–647
    [Google Scholar]
  5. Derrick P. M., Barker H., Oparka K. J. 1992; Increase in plasmodesmatal permeability during cell-to-cell spread of tobacco rattle virus from individually inoculated cells. Plant Cell 4:1405–1412
    [Google Scholar]
  6. Ding B., Haudenshield J. S., Hull R. J., Wolf S., Beachy R. N., Lucas W. J. 1992; Secondary plasmodesmata are specific sites of localization of the tobacco mosaic virus movement protein in transgenic tobacco plants. Plant Cell 4:915–928
    [Google Scholar]
  7. Fujiwara T., Giesman-Cookmeyer D., Ding B., Lommel S. A., Lucas W. J. 1993; Cell-to-cell trafficking of macromolecules through plasmodesmata potentiated by the red clover necrotic mosaic virus movement protein. Plant Cell 5:1783–1794
    [Google Scholar]
  8. Heinlein M., Epel B. L., Padgett H. S., Beachy R. N. 1995; Interaction of tobamovirus movement proteins with the plant cytoskeleton. Science 270:1983–1985
    [Google Scholar]
  9. Heinlein M., Padgett H. S., Gens J. S., Pickard B. G., Casper S. J., Epel B. L., Beachy R. N. 1998; Changing patterns of localization of the tobacco mosaic virus movement protein and replicase to the endoplasmic reticulum and microtubules during infection. Plant Cell 10:1107–1120
    [Google Scholar]
  10. Huang M., Zhang L. 1999; Association of the movement protein of alfalfa mosaic virus with the endoplasmic reticulum and its trafficking in epidermal cells of onion bulb scales. Molecular Plant–Microbe Interactions 12:680–690
    [Google Scholar]
  11. Itaya A., Hickman H., Bao Y., Nelson R., Ding B. 1997; Cell-to-cell trafficking of cucumber mosaic virus movement protein: green fluorescent protein fusion produced by biolistic gene bombardment in tobacco. Plant Journal 12:1221–1230
    [Google Scholar]
  12. Kaplan I. B., Shintaku M. H., Li Q., Zhang L., Marsh L. E., Palukaitis P. 1995; Complementation of virus movement in transgenic tobacco expressing the cucumber mosaic virus 3a gene. Virology 209:188–199
    [Google Scholar]
  13. Kasteel D. T. J., Perbal M.-C., Boyer J.-C., Wellink J., Goldbach R. W., Maule A. J., van Lent J. W. M. 1996; The movement proteins of cowpea mosaic virus and cauliflower mosaic virus induce tubular structures in plant and insect cells. Journal of General Virology 77:2857–2864
    [Google Scholar]
  14. Kasteel D. T. J., van der Wel N. N., Jansen K. A. J., Goldbach R. W., van Lent J. W. M. 1997; Tubule-forming capacity of the movement proteins of alfalfa mosaic virus and brome mosaic virus. Journal of General Virology 78:2089–2093
    [Google Scholar]
  15. Koonin E. V., Dolja V. V. 1993; Evolution and taxonomy of positive-strand RNA viruses: implications of comparative analysis of amino acid sequences. Critical Reviews in Biochemistry and Molecular Biology 28:375–430
    [Google Scholar]
  16. Lucas W. J., Gilbertson R. L. 1994; Plasmodesmata in relation to viral movement within leaf tissues. Annual Review of Phytopathology 32:387–411
    [Google Scholar]
  17. McLean B. G., Zupan J., Zambryski P. C. 1995; Tobacco mosaic virus movement protein associates with the cytoskeleton in tobacco cells. Plant Cell 7:2101–2114
    [Google Scholar]
  18. Mitsuhara I., Ugaki M., Hirochika H., Ohshima M., Murakami T., Gotoh Y., Katayose Y., Nakamura S., Honkura R., Nishimiya S., Ueno K., Mochizuki A., Tanimoto H., Tsugawa H., Otsuki Y., Ohashi Y. 1996; Efficient promoter cassettes for enhanced expression of foreign genes in dicotyledonous and monocotyledonous plants. Plant and Cell Physiology 37:49–59
    [Google Scholar]
  19. Oparka K. J., Prior D. A. M., Santa Cruz S., Padgett H. S., Beachy R. N. 1997; Gating of epidermal plasmodesmata is restricted to the leading edge of expanding infection sites of tobacco mosaic virus (TMV. Plant Journal 12:781–789
    [Google Scholar]
  20. Oparka K. J., Roberts A. G., Boevink P., Santa Cruz S., Roberts I., Pradel K. S., Imlau A., Kotlizky G., Sauer N., Epel B. 1999; Simple, but not branched, plasmodesmata allow the nonspecific trafficking of proteins in developing tobacco leaves. Cell 97:743–754
    [Google Scholar]
  21. Padgett H. S., Epel B. L., Kahn T. W., Heinlein M., Watanabe Y., Beachy R. N. 1996; Distribution of tobamovirus movement protein in infected cells and implications for cell-to-cell spread of infection. Plant Journal 10:1079–1088
    [Google Scholar]
  22. Poirson A., Turner A. P., Giovane C., Berna A., Roberts K., Godefroy-Colburn T. 1993; Effect of the alfalfa mosaic virus movement protein expressed in transgenic plants on the permeability of plasmodesmata. Journal of General Virology 74:2459–2461
    [Google Scholar]
  23. Reichel C., Más P., Beachy R. N. 1999; The role of the ER and cytoskeleton in plant viral trafficking. Trends in Plant Science 4:458–462
    [Google Scholar]
  24. Sato K., Yoshikawa N., Takahashi T. 1993; Complete nucleotide sequence of the genome of an apple isolate of apple chlorotic leaf spot virus. Journal of General Virology 74:1927–1931
    [Google Scholar]
  25. Sato K., Yoshikawa N., Takahashi T., Taira H. 1995; Expression, subcellular location and modification of the 50 kDa protein encoded by ORF2 of the apple chlorotic leaf spot trichovirus genome. Journal of General Virology 76:1503–1507
    [Google Scholar]
  26. Satoh H., Yoshikawa N., Takahashi T. 1999; Construction and biolistic inoculation of an infectious cDNA clone of apple chlorotic leaf spot trichovirus. Annals of the Phytopathological Society of Japan 65:301–304
    [Google Scholar]
  27. Storms M. M. H., Kormelink R., Peters D., van Lent J. W. M., Goldbach R. W. 1995; The nonstructural NSm protein of tomato spotted wilt virus induces tubular structures in plant and insect cells. Virology 214:485–493
    [Google Scholar]
  28. Tomenius K., Clapham D., Meshi T. 1987; Localization by immunogold cytochemistry of the virus-coded 30K protein in plasmodesmata of leaves infected with tobacco mosaic virus. Virology 160:363–371
    [Google Scholar]
  29. van Lent J., Wellink J., Goldbach R. 1990; Evidence for the involvement of the 58K and 48K proteins in the intercellular movement of cowpea mosaic virus. Journal of General Virology 71:219–223
    [Google Scholar]
  30. van Lent J., Storms M., van der Meer F., Wellink J., Goldbach R. 1991; Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts. Journal of General Virology 72:2615–2623
    [Google Scholar]
  31. Vaquero C., Turner A. P., Demangeat G., Sanz A., Serra M. T., Roberts K., Garcia-Luque I. 1994; The 3a protein from cucumber mosaic virus increases the gating capacity of plasmodesmata in transgenic tobacco plants. Journal of General Virology 75:3193–3197
    [Google Scholar]
  32. Waigmann E., Zambryski P. 1995; Tobacco mosaic virus movement protein-mediated protein transport between trichome cells. Plant Cell 7:2069–2079
    [Google Scholar]
  33. Wolf S., Deom C. M., Beachy R. N., Lucas W. J. 1989; Movement protein of tobacco mosaic virus modifies plasmodesmatal size exclusion limit. Science 246:377–379
    [Google Scholar]
  34. Yoshikawa N., Takahashi T. 1988; Properties of RNAs and proteins of apple stem grooving and apple chlorotic leaf spot viruses. Journal of General Virology 69:241–245
    [Google Scholar]
  35. Yoshikawa N., Iida H., Goto S., Magome H., Takahashi T., Terai Y. 1997; Grapevine berry inner necrosis, a new trichovirus: comparative studies with several known trichoviruses. Archives of Virology 142:1351–1363
    [Google Scholar]
  36. Yoshikawa N., Oogake S., Terada M., Miyabayashi S., Ikeda Y., Takahashi T., Ogawa K. 1999; Apple chlorotic leaf spot virus 50 kDa protein is targeted to plasmodesmata and accumulates in sieve elements in transgenic plant leaves. Archives of Virology 144:2475–2483
    [Google Scholar]
  37. Yoshikawa N., Gotoh S., Umezawa M., Satoh N., Satoh H., Takahashi T., Ito T., Yoshida K. 2000; Transgenic Nicotiana occidentalis plants expressing the 50-kDa protein of Apple chlorotic leaf spot virus display increased susceptibility to homologous virus, but strong resistance to Grapevine berry inner necrosis virus . Phytopathology 90:311–316
    [Google Scholar]
  38. Zheng H., Wang G., Zhang L. 1997; Alfalfa mosaic virus movement protein induces tubules in plant protoplasts. Molecular Plant–Microbe Interactions 10:1010–1014
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-81-8-2085
Loading
/content/journal/jgv/10.1099/0022-1317-81-8-2085
Loading

Data & Media loading...

Most cited Most Cited RSS feed