1887

Abstract

The subcellular distribution of the movement proteins (MPs) of nine alanine-scanning mutants of (CMV), fused to the green fluorescent protein (GFP) and expressed from CMV, was determined by confocal microscopy of infected epidermal cells of and , as well as infected protoplasts. Only those mutant MPs that were functional for movement in all host species tested localized to plasmodesmata of infected epidermal cells and to tubules extending from the surface of infected protoplasts, as for wild-type CMV 3a MP. Various mutant MPs that were either conditionally functional for movement or dysfunctional for movement did not localize to plasmodesmata and did not form tubules on the surface of infected protoplasts. Rather, they showed distribution to different extents throughout the infected cells, including the cytoplasm, nucleus or the plasma membrane. The CMV 3a MP also did not associate with microtubles.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.80351-0
2005-04-01
2019-11-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/86/4/vir861223.html?itemId=/content/journal/jgv/10.1099/vir.0.80351-0&mimeType=html&fmt=ahah

References

  1. Blackman, L. M., Boevink, P., Santa Cruz, S., Palukaitis, P. & Oparka, K. J. ( 1998; ). The movement protein of Cucumber mosaic virus traffics into sieve elements in minor veins of Nicotiana clevelandii. Plant Cell 10, 525–537.[CrossRef]
    [Google Scholar]
  2. Boyko, V., Ferralli, J. & Heinlein, M. ( 2000; ). Cell-to-cell movement of TMV RNA is temperature-dependent and corresponds to the association of movement protein with microtubules. Plant J 22, 315–325.[CrossRef]
    [Google Scholar]
  3. Canto, T. & Palukaitis, P. ( 1999a; ). Are tubules generated by the 3a protein necessary for cucumber mosaic virus movement? Mol Plant Microbe Interact 12, 985–993.[CrossRef]
    [Google Scholar]
  4. Canto, T. & Palukaitis, P. ( 1999b; ). The hypersensitive response to cucumber mosaic virus in Chenopodium amaranticolor requires virus movement outside the initially infected cell. Virology 265, 74–82.[CrossRef]
    [Google Scholar]
  5. Canto, T., Prior, D. A. M., Hellwald, K.-H., Oparka, K. J. & Palukaitis, P. ( 1997; ). Characterization of cucumber mosaic virus. IV. Movement protein and coat protein are both essential for cell-to-cell movement of cucumber mosaic virus. Virology 237, 237–248.[CrossRef]
    [Google Scholar]
  6. Carvalho, C. M., Pouwels, J., van Lent, J. W. M., Bisseling, T., Goldbach, R. W. & Wellink, J. ( 2004; ). The movement protein of Cowpea mosaic virus binds GTP and single-stranded nucleic acid in vitro. J Virol 78, 1591–1594.[CrossRef]
    [Google Scholar]
  7. Citovsky, V., Knorr, D. & Zambryski, P. ( 1991; ). Gene I, a potential cell-to-cell movement locus of cauliflower mosaic virus, encodes an RNA-binding protein. Proc Natl Acad Sci U S A 88, 2476–2480.[CrossRef]
    [Google Scholar]
  8. Ding, B., Li, Q., Nguyen, L., Palukaitis, P. & Lucas, W. J. ( 1995; ). Cucumber mosaic virus 3a protein potentiates cell-to-cell trafficking of CMV RNA in tobacco plants. Virology 207, 345–353.[CrossRef]
    [Google Scholar]
  9. Gillespie, T., Boevink, P., Haupt, S., Roberts, A. G., Toth, R., Valentine, T., Chapman, S. & Oparka, K. J. ( 2002; ). Functional analysis of a DNA-shuffled movement protein reveals that microtubules are dispensable for the cell-to-cell movement of Tobacco mosaic virus. Plant Cell 14, 1207–1222.[CrossRef]
    [Google Scholar]
  10. Grieco, F., Castellano, M. A., Di Sansebastiano, G. P., Maggipinto, G., Neuhaus, J.-M. & Martelli, G. P. ( 1999; ). Subcellular localization and in vivo identification of the putative movement protein of olive latent virus 2. J Gen Virol 80, 1103–1109.
    [Google Scholar]
  11. Heinlein, M., Epel, B. L., Padgett, H. S. & Beachy, R. N. ( 1995; ). Interactions of tobamovirus movement proteins with the plant cytoskeleton. Science 270, 1983–1985.[CrossRef]
    [Google Scholar]
  12. 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.[CrossRef]
    [Google Scholar]
  13. Huang, Z., Han, Y. & Howell, S. H. ( 2000; ). Formation of surface tubules and fluorescent foci in Arabidopsis thaliana protoplasts expressing a fusion between the green fluorescent protein and the cauliflower mosaic virus movement protein. Virology 271, 58–64.[CrossRef]
    [Google Scholar]
  14. Huang, M., Jongejan, L., Zheng, H., Zhang, L. & Bol, J. F. ( 2001; ). Intracellular localization and movement phenotypes of Alfalfa mosaic virus movement protein mutants. Mol Plant Microbe Interact 14, 1063–1074.[CrossRef]
    [Google Scholar]
  15. Kahn, T. W., Lapidot, M., Heinlein, M., Reichel, C., Cooper, B., Gafny, R. & Beachy, R. N. ( 1998; ). Domains of the TMV movement protein involved in subcellular localization. Plant J 15, 15–25.[CrossRef]
    [Google Scholar]
  16. 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.[CrossRef]
    [Google Scholar]
  17. Kaplan, I. B., Gal-On, A. & Palukaitis, P. ( 1997; ). Characterization of cucumber mosaic virus. III. Localization of sequences in the movement protein controlling systemic infection in cucurbits. Virology 230, 343–349.[CrossRef]
    [Google Scholar]
  18. 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. J Gen Virol 77, 2857–2864.[CrossRef]
    [Google Scholar]
  19. Kim, S. H., Kalinina, N. O., Andreev, I., Ryabov, E. V., Fitzgerald, A. G., Taliansky, M. E. & Palukaitis, P. ( 2004; ). The C-terminal 33 amino acids of the cucumber mosaic virus 3a protein affect virus movement, RNA binding and inhibition of infection and translation. J Gen Virol 85, 221–230.[CrossRef]
    [Google Scholar]
  20. Kragler, F., Curin, M., Trutnyeva, K., Gansch, A. & Waigmann, E. ( 2003; ). MPB2C, a microtubule-associated plant protein binds to and interferes with cell-to-cell transport of tobacco mosaic virus movement protein. Plant Physiol 132, 1870–1883.[CrossRef]
    [Google Scholar]
  21. Laporte, C., Vetter, G., Loudes, A.-M., Robinson, D. G., Hillmer, S., Stussi-Garaud, C. & Ritzenthaler, C. ( 2003; ). Involvement of the secretory pathway and the cytoskeleton in the intracellular targeting and tubule assembly of Grapevine fanleaf virus movement protein in tobacco BY-2 cells. Plant Cell 15, 2058–2075.[CrossRef]
    [Google Scholar]
  22. Li, Q. & Palukaitis, P. ( 1996; ). Comparison of the nucleic acid- and NTP-binding properties of the movement protein of cucumber mosaic cucumovirus and tobacco mosaic tobamovirus. Virology 216, 71–79.[CrossRef]
    [Google Scholar]
  23. Li, Q., Ryu, K. H. & Palukaitis, P. ( 2001; ). Cucumber mosaic virus–plant interactions: identification of 3a protein sequences affecting infectivity, cell-to-cell movement, and long-distance movement. Mol Plant Microbe Interact 14, 378–385.[CrossRef]
    [Google Scholar]
  24. Linstead, P. J., Hills, G. J., Plaskitt, K. A., Wilson, I. G., Harker, C. L. & Maule, A. J. ( 1988; ). The subcellular location of the gene 1 product of cauliflower mosaic virus is consistent with a function associated with virus spread. J Gen Virol 69, 1809–1818.[CrossRef]
    [Google Scholar]
  25. 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.[CrossRef]
    [Google Scholar]
  26. Nagano, H., Okuno, T., Mise, K. & Furusawa, I. ( 1997; ). Deletion of the C-terminal 33 amino acids of cucumber mosaic virus movement protein enables a chimeric brome mosaic virus to move from cell to cell. J Virol 71, 2270–2276.
    [Google Scholar]
  27. Palukaitis, P. & García-Arenal, F. ( 2003; ). Cucumoviruses. Adv Virus Res 62, 241–323.
    [Google Scholar]
  28. Pouwels, J., van der Krogt, G. N. M., van Lent, J., Bisseling, T. & Wellink, J. ( 2002; ). The cytoskeleton and the secretory pathway are not involved in targeting the cowpea mosaic virus movement protein to the cell periphery. Virology 297, 48–56.[CrossRef]
    [Google Scholar]
  29. Pouwels, J., Kornet, N., van Bers, N., Guighelaar, T., van Lent, J., Bisseling, T. & Wellink, J. ( 2003; ). Identification of distinct steps during tubule formation by the movement protein of Cowpea mosaic virus. J Gen Virol 84, 3485–3494.[CrossRef]
    [Google Scholar]
  30. Schoumacher, F., Erny, C., Berna, A., Godefroy-Colburn, T. & Stussi-Garaud, C. ( 1992; ). Nucleic acid-binding properties of the alfalfa mosaic virus movement protein produced in yeast. Virology 188, 896–899.[CrossRef]
    [Google Scholar]
  31. Takeshita, M., Suzuki, M. & Takanami, Y. ( 2001; ). Combination of amino acids in the 3a protein and the coat protein of Cucumber mosaic virus determines symptom expression and viral spread in bottle gourd. Arch Virol 146, 697–711.[CrossRef]
    [Google Scholar]
  32. Ueda, K., Matsuyama, T. & Hashimoto, T. ( 1999; ). Visualization of microtubules in living cells of transgenic Arabidopsis thaliana. Protoplasma 206, 201–206.[CrossRef]
    [Google Scholar]
  33. 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. J Gen Virol 7271, 219–223.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.80351-0
Loading
/content/journal/jgv/10.1099/vir.0.80351-0
Loading

Data & Media loading...

Supplements

vol. , part 4, pp. 1223 – 1228

Localization of MP–red fluorescent protein and tubulin–GFP in epidermal cells. Genes encoding the wild-type (WT) 3a protein, as well as the MPs of the dysfunctional mutant M4 and the conditionally functional mutant M8, were cloned in the binary vector pROK2 and fused at their C termini to the monomeric red fluorescent protein (mRFP; Campbell , 2002). These binary constructs were agroinfiltrated into either transgenic plants expressing α-tubulin fused to GFP ( –GFP) (line CB13; Gillespie , 2002) for transient expression of the tagged MPs (WT and M4), or non-transgenic co-infiltrated with a binary construct expressing –GFP (Ueda , 1999) (M8). At 3 days post-infiltration, the three MP–mRFP fusions showed the same intracellular distribution of fluorescence as when tagged with GFP: discrete spots at the cell wall for WT 3a–mRFP (upper panels) or cytoplasmic and nuclear distribution for both M4–mRFP and M8–mRFP, the former showing a more granular and threadbare appearance than the latter (middle and lower panels, respectively). In neither case did the red fluorescence appear to target the microtubule filaments highlighted green by –GFP. Left panels show whole cells; right panels show enhanced detail of the intracellular microtubular net. Bars, 50 µm.

A monomeric red fluorescent protein. , 7877–7882.

Functional analysis of a DNA-shuffled movement protein reveals that microtubules are dispensable for the cell-to-cell movement of . , 1207–1222.

Visualization of microtubules in living cells of transgenic . , 201–206.



IMAGE

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error