1887

Abstract

The 2b protein encoded by (CMV) has been shown to be a silencing suppressor and pathogenicity determinant in solanaceous hosts, but a movement determinant in cucumber. In addition, synergistic interactions between CMV and (ZYMV) have been described in several cucurbit species. Here, it was shown that deletion of the gene from CMV prevented extensive systemic movement of the virus in zucchini squash, which could not be complemented by co-infection with ZYMV. Thus, ZYMV expressing a silencing suppressor with a different target could not complement the CMV 2b-specific movement function. Expression of the 2b protein from an attenuated ZYMV vector resulted in a synergistic response, largely restoring infection symptoms of wild-type ZYMV in several cucurbit species. Deletion or alteration of either of two nuclear localization signals (NLSs) did not affect nuclear localization in two assays, but did affect pathogenicity in several cucurbit species, whilst deletion of both NLSs led to loss of nuclear localization. The 2b protein interacted with an karyopherin protein (AtKAP) in the yeast two-hybrid system, as did each of the two single NLS-deletion mutants. However, 2b protein containing a deletion of both NLSs was unable to interact with AtKAP. These data suggest that the 2b protein localizes to the nucleus by using the karyopherin -mediated system, but demonstrate that nuclear localization was insufficient for enhancement of the 2b-mediated pathogenic response in cucurbit hosts. Thus, the sequences corresponding to the two NLSs must have another role leading to pathogenicity enhancement.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.80250-0
2004-10-01
2019-11-17
Loading full text...

Full text loading...

/deliver/fulltext/jgv/85/10/vir853135.html?itemId=/content/journal/jgv/10.1099/vir.0.80250-0&mimeType=html&fmt=ahah

References

  1. Anjos, J. R., Jarlfors, U. & Ghabrial, S. A. ( 1992; ). Soybean mosaic potyvirus enhances the titer of two comoviruses in dually infected soybean plants. Phytopathology 82, 1022–1027.[CrossRef]
    [Google Scholar]
  2. Arazi, T., Slutsky, S. G., Shiboleth, Y. M., Wang, Y., Rubinstein, M., Barak, S., Yang, J. & Gal-On, A. ( 2001; ). Engineering zucchini yellow mosaic potyvirus as a non-pathogenic vector for expression of heterologous proteins in cucurbits. J Biotechnol 87, 67–82.[CrossRef]
    [Google Scholar]
  3. Ballas, N. & Citovsky, V. ( 1997; ). Nuclear localization signal binding protein from Arabidopsis mediates nuclear import of Agrobacterium VirD2 protein. Proc Natl Acad Sci U S A 94, 10723–10728.[CrossRef]
    [Google Scholar]
  4. Bennett, C. W. ( 1952; ). Interactions between viruses and virus strains. Adv Virus Res 1, 39–67.
    [Google Scholar]
  5. Brigneti, G., Voinnet, O., Li, W.-X., Ji, L.-H., Ding, S.-W. & Baulcombe, D. C. ( 1998; ). Viral pathogenicity determinants are suppressors of transgene silencing in Nicotiana benthamiana. EMBO J 17, 6739–6746.[CrossRef]
    [Google Scholar]
  6. Choi, S. K., Yoon, J. Y., Ryu, K. H., Choi, J. K., Palukaitis, P. & Park, W. M. ( 2002; ). Systemic movement of a movement-deficient strain of Cucumber mosaic virus in zucchini squash is facilitated by a cucurbit-infecting potyvirus. J Gen Virol 83, 3173–3178.
    [Google Scholar]
  7. Ding, S.-W., Li, W.-X. & Symons, R. H. ( 1995a; ). A novel naturally occurring hybrid gene encoded by a plant RNA virus facilitates long distance virus movement. EMBO J 14, 5762–5772.
    [Google Scholar]
  8. Ding, S.-W., Rathjen, J. P., Li, W.-X., Swanson, R., Healy, H. & Symons, R. H. ( 1995b; ). Efficient infection from cDNA clones of cucumber mosaic cucumovirus RNAs in a new plasmid vector. J Gen Virol 76, 459–464.[CrossRef]
    [Google Scholar]
  9. Ding, S.-W., Shi, B.-J., Li, W.-X. & Symons, R. H. ( 1996; ). An interspecies hybrid RNA virus is significantly more virulent than either parental virus. Proc Natl Acad Sci U S A 93, 7470–7474.[CrossRef]
    [Google Scholar]
  10. Fukumoto, F., Masuda, Y. & Hanada, K. ( 2003; ). Pea tissue necrosis induced by Cucumber mosaic virus alone or together with Watermelon mosaic virus. Plant Dis 87, 324–328.[CrossRef]
    [Google Scholar]
  11. Gal-On, A. ( 2000; ). A point mutation in the FRNK motif of the potyvirus helper component-protease gene alters symptom expression in cucurbits and elicits protection against the severe homologous virus. Phytopathology 90, 467–473.[CrossRef]
    [Google Scholar]
  12. Gal-On, A., Antignus, Y., Rosner, A. & Raccah, B. ( 1992; ). A zucchini yellow mosaic virus coat protein gene mutation restores aphid transmissibility but has no effect on multiplication. J Gen Virol 73, 2183–2187.[CrossRef]
    [Google Scholar]
  13. Gal-On, A., Meiri, E., Elman, C., Gray, D. J. & Gaba, V. ( 1997; ). Simple hand-held devices for the efficient infection of plants with viral-encoding constructs by particle bombardment. J Virol Methods 64, 103–110.[CrossRef]
    [Google Scholar]
  14. Garces-Orejuela, C. & Pound, G. S. ( 1957; ). The multiplication of tobacco mosaic virus in the presence of cucumber mosaic virus or tobacco ringspot virus in tobacco. Phytopathology 47, 232–239.
    [Google Scholar]
  15. González-Jara, P., Tenllado, F., Martínez-García, B., Atencio, F. A., Barajas, D., Vargas, M., Díaz-Ruíz, J. & Díaz-Ruíz, J. R. ( 2004; ). Host-dependent differences during synergistic infection by Potyviruses with potato virus X. Mol Plant Pathol 5, 29–35.[CrossRef]
    [Google Scholar]
  16. Goodin, M. M., Austin, J., Tobias, R., Fujita, M., Morales, C. & Jackson, A. O. ( 2001; ). Interactions and nuclear import of the N and P proteins of sonchus yellow net virus, a plant nucleorhabdovirus. J Virol 75, 9393–9406.[CrossRef]
    [Google Scholar]
  17. Görlich, D. & Mattaj, I. W. ( 1996; ). Nucleocytoplasmic transport. Science 271, 1513–1518.[CrossRef]
    [Google Scholar]
  18. Hollenberg, S. M., Sternglanz, R., Cheng, P. F. & Weintraub, H. ( 1995; ). Identification of a new family of tissue-specific basic helix-loop-helix proteins with a two-hybrid system. Mol Cell Biol 15, 3813–3822.
    [Google Scholar]
  19. Hunter, P. J., Jones, J. E. & Walsh, J. A. ( 2002; ). Involvement of Beet western yellows virus, Cauliflower mosaic virus, and Turnip mosaic virus in internal disorders of stored white cabbage. Phytopathology 92, 816–826.[CrossRef]
    [Google Scholar]
  20. Ji, L.-H. & Ding, S.-W. ( 2001; ). The suppressor of transgene RNA silencing encoded by Cucumber mosaic virus interferes with salicylic acid-mediated virus resistance. Mol Plant Microbe Interact 14, 715–724.[CrossRef]
    [Google Scholar]
  21. Karyeija, R. F., Kreuze, J. F., Gibson, R. W. & Valkonen, J. P. T. ( 2000; ). Synergistic interactions of a potyvirus and a phloem-limited crinivirus in sweet potato plants. Virology 269, 26–36.[CrossRef]
    [Google Scholar]
  22. Kasschau, K. D., Xie, Z., Allen, E., Llave, C., Chapman, E. J., Krizan, K. A. & Carrington, J. C. ( 2003; ). P1/HC-Pro, a viral suppressor of RNA silencing, interferes with Arabidopsis development and miRNA function. Dev Cell 4, 205–217.[CrossRef]
    [Google Scholar]
  23. Kunik, T., Palanichelvam, K., Czosnek, H., Citovsky, V. & Gafni, Y. ( 1998; ). Nuclear import of the capsid protein of tomato yellow leaf curl virus (TYLCV) in plant and insect cells. Plant J 13, 393–399.[CrossRef]
    [Google Scholar]
  24. Kunkel, T. A., Roberts, J. D. & Zakour, R. A. ( 1987; ). Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol 154, 367–382.
    [Google Scholar]
  25. Leclerc, D., Chapdelaine, Y. & Hohn, T. ( 1999; ). Nuclear targeting of the cauliflower mosaic virus coat protein. J Virol 73, 553–560.
    [Google Scholar]
  26. Li, H.-W., Lucy, A. P., Guo, H.-S., Li, W.-X., Ji, L.-H., Wong, S.-M. & Ding, S.-W. ( 1999; ). Strong host resistance targeted against a viral suppressor of the plant gene silencing defence mechanism. EMBO J 18, 2683–2691.[CrossRef]
    [Google Scholar]
  27. Lucy, A. P., Guo, H.-S., Li, W.-X. & Ding, S.-W. ( 2000; ). Suppression of post-transcriptional gene silencing by a plant viral protein localized in the nucleus. EMBO J 19, 1672–1680.[CrossRef]
    [Google Scholar]
  28. Mayers, C. N., Palukaitis, P. & Carr, J. P. ( 2000; ). Subcellular distribution analysis of the cucumber mosaic virus 2b protein. J Gen Virol 81, 219–226.
    [Google Scholar]
  29. Murphy, J. F. & Kyle, M. M. ( 1995; ). Alleviation of restricted systemic spread of pepper mottle potyvirus in Capsicum annuum cv. Avelar by coinfection with a cucumovirus. Phytopathology 85, 561–566.[CrossRef]
    [Google Scholar]
  30. Palukaitis, P. & Kaplan, I. B. ( 1997; ). Synergy of virus accumulation and pathology in transgenic plants expressing viral sequences. In Virus-Resistant Transgenic Plants: Potential Ecological Impact, pp. 77–84. Edited by M. Tepfer & E. Balázs. Berlin: Springer.
  31. Pio-Ribeiro, G., Wyatt, S. D. & Kuhn, C. W. ( 1978; ). Cowpea stunt: a disease caused by a synergistic interaction of two viruses. Phytopathology 68, 1260–1265.[CrossRef]
    [Google Scholar]
  32. Poolpol, P. & Inouye, T. ( 1986; ). Enhancement of cucumber mosaic virus multiplication by zucchini yellow mosaic virus in doubly infected cucumber plants. Ann Phytopathol Soc Jpn 52, 22–30.[CrossRef]
    [Google Scholar]
  33. Pruss, G., Ge, X., Shi, X. M., Carrington, J. C. & Bowman Vance, V. ( 1997; ). Plant viral synergism: the potyviral genome encodes a broad-range pathogenicity enhancer that transactivates replication of heterologous viruses. Plant Cell 9, 859–868.[CrossRef]
    [Google Scholar]
  34. Qiu, W., Park, J.-W. & Scholthof, H. B. ( 2002; ). Tombusvirus P19-mediated suppression of virus-induced gene silencing is controlled by genetic and dosage features that influence pathogenicity. Mol Plant Microbe Interact 15, 269–280.[CrossRef]
    [Google Scholar]
  35. Rhee, Y., Gurel, F., Gafni, Y., Dingwall, C. & Citovsky, V. ( 2000; ). A genetic system for detection of protein nuclear import and export. Nat Biotechnol 18, 433–437.[CrossRef]
    [Google Scholar]
  36. Rizzo, T. M. & Palukaitis, P. ( 1990; ). Construction of full-length cDNA clones of cucumber mosaic virus RNAs 1, 2 and 3: generation of infectious RNA transcripts. Mol Gen Genet 222, 249–256.[CrossRef]
    [Google Scholar]
  37. Rochow, W. F. & Ross, A. F. ( 1955; ). Virus multiplication in plants doubly infected by potato viruses X and Y. Virology 1, 10–27.[CrossRef]
    [Google Scholar]
  38. Ryabov, E. V., Fraser, G., Mayo, M. A., Barker, H. & Taliansky, M. ( 2001; ). Umbravirus gene expression helps Potato leafroll virus to invade mesophyll tissues and to be transmitted mechanically between plants. Virology 286, 363–372.[CrossRef]
    [Google Scholar]
  39. Sáenz, P., Quiot, L., Quiot, J.-B., Candresse, T. & García, J. A. ( 2001; ). Pathogenicity determinants in the complex virus population of a Plum pox virus isolate. Mol Plant Microbe Interact 14, 278–287.[CrossRef]
    [Google Scholar]
  40. Sáenz, P., Salvador, B., Simón-Mateo, C., Kasschau, K. D., Carrington, J. C. & García, J. A. ( 2002; ). Host-specific involvement of the HC protein in the long-distance movement of potyviruses. J Virol 76, 1922–1931.[CrossRef]
    [Google Scholar]
  41. Sano, Y. & Kojima, M. ( 1989; ). Increase in cucumber mosaic virus concentration in Japanese radish plants co-infected with turnip mosaic virus. Ann Phytopathol Soc Jpn 55, 296–302.[CrossRef]
    [Google Scholar]
  42. Selth, L. A., Randles, J. W. & Rezaian, M. A. ( 2004; ). Host responses to transient expression of individual genes encoded by Tomato leaf curl virus. Mol Plant Microbe Interact 17, 27–33.[CrossRef]
    [Google Scholar]
  43. SenGupta, D. J., Zhang, B., Kraemer, B., Pochart, P., Fields, S. & Wickens, M. ( 1996; ). A three-hybrid system to detect RNA–protein interactions in vivo. Proc Natl Acad Sci U S A 93, 8496–8501.[CrossRef]
    [Google Scholar]
  44. Shi, B.-J., Palukaitis, P. & Symons, R. H. ( 2002; ). Differential virulence by strains of Cucumber mosaic virus is mediated by the 2b gene. Mol Plant Microbe Interact 15, 947–955.[CrossRef]
    [Google Scholar]
  45. Shi, B.-J., Miller, J., Symons, R. H. & Palukaitis, P. ( 2003; ). The 2b protein of cucumoviruses has a role in promoting the cell-to-cell movement of pseudorecombinant viruses. Mol Plant Microbe Interact 16, 261–267.[CrossRef]
    [Google Scholar]
  46. Soards, A. J., Murphy, A. M., Palukaitis, P. & Carr, J. P. ( 2002; ). Virulence and differential local and systemic spread of Cucumber mosaic virus in tobacco are affected by the CMV 2b protein. Mol Plant Microbe Interact 15, 647–653.[CrossRef]
    [Google Scholar]
  47. Stachel, S. E., An, G., Flores, C. & Nester, E. W. ( 1985; ). A Tn3 lacZ transposon for the random generation of beta-galactosidase gene fusions: application to the analysis of gene expression in Agrobacterium. EMBO J 4, 891–898.
    [Google Scholar]
  48. Suntharalingam, M. & Wente, S. R. ( 2003; ). Peering through the pore: nuclear pore complex structure, assembly, and function. Dev Cell 4, 775–789.[CrossRef]
    [Google Scholar]
  49. Sutton, A., Heller, R. C., Landry, J., Choy, J. S., Sirko, A. & Sternglanz, R. ( 2001; ). A novel form of transcriptional silencing by Sum1-1 requires Hst1 and the origin recognition complex. Mol Cell Biol 21, 3514–3522.[CrossRef]
    [Google Scholar]
  50. Tzfira, T. & Citovsky, V. ( 2001; ). Comparison between nuclear localization of nopaline- and octopine-specific Agrobacterium VirE2 proteins in plant, yeast and mammalian cells. Mol Plant Pathol 2, 171–176.[CrossRef]
    [Google Scholar]
  51. Tzfira, T., Vaidya, M. & Citovsky, V. ( 2001; ). VIP1, an Arabidopsis protein that interacts with Agrobacterium VirE2, is involved in VirE2 nuclear import and Agrobacterium infectivity. EMBO J 20, 3596–3607.[CrossRef]
    [Google Scholar]
  52. Tzfira, T., Vaidya, M. & Citovsky, V. ( 2002; ). Increasing plant susceptibility to Agrobacterium infection by overexpression of the Arabidopsis nuclear protein VIP1. Proc Natl Acad Sci U S A 99, 10435–10440.[CrossRef]
    [Google Scholar]
  53. van Bel, A. J. E. ( 1993; ). Strategies of phloem loading. Annu Rev Plant Physiol Plant Mol Biol 44, 253–281.[CrossRef]
    [Google Scholar]
  54. Varagona, M. J., Schmidt, R. J. & Raikhel, N. V. ( 1992; ). Nuclear localization signal(s) required for nuclear targeting of the maize regulatory protein Opaque-2. Plant Cell 4, 1213–1227.[CrossRef]
    [Google Scholar]
  55. Wang, Y., Gaba, V., Yang, J., Palukaitis, P. & Gal-On, A. ( 2002; ). Characterization of synergy between Cucumber mosaic virus and potyviruses in cucurbit hosts. Phytopathology 92, 51–58.[CrossRef]
    [Google Scholar]
  56. Wang, Y., Lee, K. C., Gaba, V., Wong, S. M., Palukaitis, P. & Gal-On, A. ( 2004; ). Breakage of resistance to Cucumber mosaic virus by co-infection with Zucchini mosaic virus: enhancement of CMV accumulation independent of symptom expression. Arch Virol 149, 379–396.[CrossRef]
    [Google Scholar]
  57. Zhang, L., Handa, K. & Palukaitis, P. ( 1994; ). Mapping local and systemic symptom determinants of cucumber mosaic cucumovirus in tobacco. J Gen Virol 75, 3185–3191.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.80250-0
Loading
/content/journal/jgv/10.1099/vir.0.80250-0
Loading

Data & Media loading...

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