1887

Abstract

Potyviruses are non-persistently transmitted by aphid vectors with the assistance of a viral accessory factor known as helper component (HC-Pro), a multifunctional protein that is also involved in many other essential processes during the virus infection cycle. A transient -mediated expression system was used to produce Plum pox virus (PPV) HC-Pro in leaves from constructs that incorporated the 5′ region of the genome, yielding high levels of HC-Pro in agroinfiltrated leaves. The expressed PPV HC-Pro was able to assist aphid transmission of purified virus particles in a sequential feeding assay, and to complement transmission-defective variants of the virus. Also, HC-Pro of a second potyvirus, (TEV), was expressed and found to be functional for aphid transmission. These results show that this transient system can be useful for production of functionally active HC-Pro in potyviruses, and the possible uses of this approach to study the mechanism of transmission are discussed.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.82301-0
2006-11-01
2020-10-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/87/11/3413.html?itemId=/content/journal/jgv/10.1099/vir.0.82301-0&mimeType=html&fmt=ahah

References

  1. Allison R., Johnston R. E., Dougherty W. G. 1986; The nucleotide sequence of the coding region of tobacco etch virus genomic RNA: evidence for the synthesis of a single polyprotein. Virology 154:9–20 [CrossRef]
    [Google Scholar]
  2. Anandalakshmi R., Pruss G. J., Ge X., Marathe R., Mallory A. C., Smith T. H., Vance V. B. 1998; A viral suppressor of gene silencing in plants. Proc Natl Acad Sci U S A 95:13079–13084 [CrossRef]
    [Google Scholar]
  3. Andrews L. B., Curtis W. R. 2005; Comparison of transient protein expression in tobacco leaves and plant suspension culture. Biotechnol Prog 21:946–952
    [Google Scholar]
  4. Atreya C. D., Raccah B., Pirone T. P. 1990; A point mutation in the coat protein abolishes aphid transmissibility of a potyvirus. Virology 178:161–165 [CrossRef]
    [Google Scholar]
  5. Barajas D., Tenllado F., Gonzalez-Jara P., Martinez-Garcia B., Atencio F. A., Díaz-Ruíz J. R. 2004; Resistance to plum pox virus (PPV) in Nicotiana benthamiana plants transformed with the PPV HC-Pro silencing suppressor gene. J Plant Pathol 86:239–248
    [Google Scholar]
  6. Berger P. H., Hunt A. G., Domier L. L., Hellmann G. M., Stram Y., Thornbury D. W., Pirone T. P. 1989; Expression in transgenic plants of a viral gene product that mediates insect transmission of potyviruses. Proc Natl Acad Sci U S A 86:8402–8406 [CrossRef]
    [Google Scholar]
  7. Blanc S., Lopez-Moya J. J., Wang R., Garcia-Lampasona S., Thornbury D. W., Pirone T. P. 1997; A specific interaction between coat protein and helper component correlates with aphid transmission of a potyvirus. Virology 231:141–147 [CrossRef]
    [Google Scholar]
  8. Blanc S., Ammar E. D., Garcia-Lampasona S., Dolja V. V., Llave C., Baker J., Pirone T. P. 1998; Mutations in the potyvirus helper component protein: effects on interactions with virions and aphid stylets. J Gen Virol 79:3119–3122
    [Google Scholar]
  9. Blanc S., Dolja V., Llave C., Pirone T. 1999; Histidine-tagging and purification of tobacco etch potyvirus helper component protein. J Virol Methods 77:11–15 [CrossRef]
    [Google Scholar]
  10. 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]
  11. Canto T., Lopez-Moya J. J., Serra-Yoldi M. T., Diaz Ruiz J. R., Lopez-Abella D. 1995; Different helper component mutations associated with lack of aphid transmissibility in two isolates of potato virus Y. Phytopathology 85:1519–1524 [CrossRef]
    [Google Scholar]
  12. Carrington J. C., Cary S. M., Parks T. D., Dougherty W. G. 1989; A second proteinase encoded by a plant potyvirus genome. EMBO J 8:365–370
    [Google Scholar]
  13. Cronin S., Verchot J., Haldeman-Cahill R., Schaad M. C., Carrington J. C. 1995; Long-distance movement factor: a transport function of the potyvirus helper component proteinase. Plant Cell 7:549–559 [CrossRef]
    [Google Scholar]
  14. Dolja V. V., McBride H. J., Carrington J. C. 1992; Tagging of plant potyvirus replication and movement by insertion of beta-glucuronidase into the viral polyprotein. Proc Natl Acad Sci U S A 89:10208–10212 [CrossRef]
    [Google Scholar]
  15. Dolja V. V., Herndon K. L., Pirone T. P., Carrington J. C. 1993; Spontaneous mutagenesis of a plant potyvirus genome after insertion of a foreign gene. J Virol 67:5968–5975
    [Google Scholar]
  16. Earley K. W., Haag J. R., Pontes O., Opper K., Juehne T., Song K., Pikaard C. S. 2006; Gateway-compatible vectors for plant functional genomics and proteomics. Plant J 45:616–629 [CrossRef]
    [Google Scholar]
  17. English J. J., Davenport G. F., Elmayan T., Vaucheret H., Baulcombe D. 1997; Requirement of sense transcription for homology-dependent virus resistance and trans-inactivation. Plant J 12:597–603 [CrossRef]
    [Google Scholar]
  18. Fernandez-Fernandez M. R., Martinez-Torrecuadrada J. L., Roncal F., Dominguez E., Garcia J. A. 2002; Identification of immunogenic hot spots within plum pox potyvirus capsid protein for efficient antigen presentation. J Virol 76:12646–12653 [CrossRef]
    [Google Scholar]
  19. Fischer R., Vaquero-Martin C., Sack M., Drossard J., Emans N., Commandeur U. 1999; Towards molecular farming in the future: transient protein expression in plants. Biotechnol Appl Biochem 30:113–116
    [Google Scholar]
  20. Gonzalez-Jara P., Atencio F. A., Martinez-Garcia B., Barajas D., Tenllado F., Diaz-Ruiz J. R. 2005; A single amino acid mutation in the plum pox virus helper component-proteinase gene abolishes both synergistic and RNA silencing suppression activities. Phytopathology 95:894–901 [CrossRef]
    [Google Scholar]
  21. Govier D. A., Kassanis B. 1974a; Evidence that a component other than the virus particle is needed for aphid transmission of potato virus Y. Virology 57:285–286 [CrossRef]
    [Google Scholar]
  22. Govier D. A., Kassanis B. 1974b; A virus-induced component of plant sap needed when aphids acquire potato virus Y from purified preparations. Virology 61:420–426 [CrossRef]
    [Google Scholar]
  23. Guo H. S., Lopez-Moya J. J., Garcia J. A. 1998; Susceptibility to recombination rearrangements of a chimaeric plum pox potyvirus genome after insertion of a foreign gene. Virus Res 57:183–195 [CrossRef]
    [Google Scholar]
  24. Johansen L. K., Carrington J. C. 2001; Silencing on the spot. Induction and suppression of RNA silencing in the Agrobacterium -mediated transient expression system. Plant Physiol 126:930–938 [CrossRef]
    [Google Scholar]
  25. Kapila J., De Rycke M. V. M., Angenon G. 1997; An Agrobacterium -mediated transient gene expression system for intact leaves. Plant Sci 122:101–108 [CrossRef]
    [Google Scholar]
  26. Kasschau K. D., Carrington J. C. 1995; Requirement for HC-Pro processing during genome amplification of tobacco etch potyvirus. Virology 209:268–273 [CrossRef]
    [Google Scholar]
  27. Kasschau K. D., Carrington J. C. 1998; A counterdefensive strategy of plant viruses: suppression of posttranscriptional gene silencing. Cell 95:461–470 [CrossRef]
    [Google Scholar]
  28. Kasschau K. D., Cronin S., Carrington J. C. 1997; Genome amplification and long-distance movement functions associated with the central domain of tobacco etch potyvirus helper component-proteinase. Virology 228:251–262 [CrossRef]
    [Google Scholar]
  29. Lain S., Riechmann J. L., Mendez E., Garcia J. A. 1988; Nucleotide sequence of the 3′ terminal region of plum pox potyvirus RNA. Virus Res 10:325–342 [CrossRef]
    [Google Scholar]
  30. Lain S., Riechmann J. L., Garcia J. A. 1989; The complete nucleotide sequence of plum pox potyvirus RNA. Virus Res 13:157–172 [CrossRef]
    [Google Scholar]
  31. Lakatos L., Csorba T., Pantaleo V. & 7 other authors 2006; Small RNA binding is a common strategy to suppress RNA silencing by several viral suppressors. EMBO J 25:2768–2780 [CrossRef]
    [Google Scholar]
  32. Llave C., Kasschau K., Carrington J. C. 2000; Virus-encoded suppressor of posttranscriptional gene silencing targets a maintenance step in the silencing pathway. Proc Natl Acad Sci U S A 21:13401–13406
    [Google Scholar]
  33. Llave C., Martinez B., Diaz-Ruiz J. R., Lopez-Abella D. 2002; Amino acid substitutions within the Cys-rich domain of the tobacco etch potyvirus HC-Pro result in loss of transmissibility by aphids. Arch Virol 147:2365–2375 [CrossRef]
    [Google Scholar]
  34. Lopez-Moya J. J., Garcia J. A. 2000; Construction of a stable and highly infectious intron-containing cDNA clone of plum pox potyvirus and its use to infect plants by particle bombardment. Virus Res 68:99–107 [CrossRef]
    [Google Scholar]
  35. Lopez-Moya J. J., Canto T., Diaz-Ruiz J. R., Lopez-Abella D. 1995; Transmission by aphids of a naturally non-transmissible plum pox virus isolate with the aid of potato virus Y helper component. J Gen Virol 76:2293–2297 [CrossRef]
    [Google Scholar]
  36. Lopez-Moya J. J., Wang R. Y., Pirone T. P. 1999; Context of the coat protein DAG motif affects potyvirus transmissibility by aphids. J Gen Virol 80:3281–3288
    [Google Scholar]
  37. Maiss E., Timpe U., Brisske A., Jelkmann W., Casper R., Himmler G., Mattanovich D., Katinger H. W. D. 1989; The complete nucleotide sequence of plum pox virus RNA. J Gen Virol 70:513–524 [CrossRef]
    [Google Scholar]
  38. Marillonnet S., Giritch A., Gils M., Kandzia R., Klimyuk V., Gleba Y. 2004; In planta engineering of viral RNA replicons: efficient assembly by recombination of DNA modules delivered by agrobacterium. Proc Natl Acad Sci U S A 101:6852–6857 [CrossRef]
    [Google Scholar]
  39. Martinez-Garcia B. 2000; Análisis molecular de las proteínas virales implicadas en la transmisión por pulgones del virus de la sharka (Plum pox virus) . PhD thesis Departamento de Biología Vegetal, Facultad de Ciencias Biológicas; Universidad Complutense de Madrid, Spain:
  40. Murphy J. F., Rhoads R. E., Hunt A. G., Shaw J. G. 1990; The VPg of tobacco etch virus RNA is the 49-kDa proteinase or the N-terminal 24-kDa part of the proteinase. Virology 178:285–288 [CrossRef]
    [Google Scholar]
  41. Orzaez D., Mirabel S., Wieland W. H., Granell A. 2006; Agroinjection of tomato fruits. A tool for rapid functional analysis of transgenes directly in fruit. Plant Physiol 140:3–11
    [Google Scholar]
  42. Peng Y.-H., Kadoury D., Gal-On A., Huet H., Wang Y., Raccah B. 1998; Mutations in the HC-Pro gene of zucchini yellow mosaic potyvirus: effects on aphid transmission and binding to purified virions. J Gen Virol 79:897–904
    [Google Scholar]
  43. Pirone T. P., Blanc S. 1996; Helper-dependent vector transmission of plant viruses. Annu Rev Phytopathol 34:227–247 [CrossRef]
    [Google Scholar]
  44. Plisson C., Drucker M., Blanc S., German-Retana S., Le Gall O., Thomas D., Bron P. 2003; Structural characterization of HC-Pro, a plant virus multifunctional protein. J Biol Chem 278:23753–23761 [CrossRef]
    [Google Scholar]
  45. 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]
  46. Rajamaki M. L., Kelloniemi J., Alminaite A., Kekarainen T., Rabenstein F., Valkonen J. P. 2005; A novel insertion site inside the potyvirus P1 cistron allows expression of heterologous proteins and suggests some P1 functions. Virology 342:88–101 [CrossRef]
    [Google Scholar]
  47. Robaglia C., Durand-Tardif M., Tronchet M., Boudazin G., Astier-Manifacier S., Casse-Delbart F. 1989; Nucleotide sequence of potato virus Y (N strain) genomic RNA. J Gen Virol 70:935–947 [CrossRef]
    [Google Scholar]
  48. Ruiz-Ferrer V., Goytia E., Martinez-Garcia B., Lopez-Abella D., Lopez-Moya J. J. 2004; Expression of functionally active helper component protein of Tobacco etch potyvirus in the yeast Pichia pastoris . J Gen Virol 85:241–249 [CrossRef]
    [Google Scholar]
  49. Saenz P., Salvador B., Simon-Mateo C., Kasschau K., Carrington J., Garcia 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]
  50. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  51. Sasaya T., Torrance L., Cowan G., Ziegler A. 2000; Aphid transmission studies using helper component proteins of Potato virus Y expressed from a vector derived from Potato virus X . J Gen Virol 81:1115–1119
    [Google Scholar]
  52. Simon-Buela L., Guo H. S., Garcia J. A. 1997; Long sequences in the 5′ noncoding region of plum pox virus are not necessary for viral infectivity but contribute to viral competitiveness and pathogenesis. Virology 233:157–162 [CrossRef]
    [Google Scholar]
  53. Thornbury D. W., Hellman G. M., Rhodes R. E., Pirone T. P. 1985; Purification and characterization of potyvirus helper component. Virology 144:260–267 [CrossRef]
    [Google Scholar]
  54. Thornbury D. W., van den Heuvel J. F., Lesnaw J. A., Pirone T. P. 1993; Expression of potyvirus proteins in insect cells infected with a recombinant baculovirus. J Gen Virol 74:2731–2735 [CrossRef]
    [Google Scholar]
  55. Torrance L., Andreev I. A., Gabrenaite-Verhovskaya R., Cowan G., Makinen K., Taliansky M. E. 2005; An unusual structure at one end of potato potyvirus particles. J Mol Biol 357:1–8
    [Google Scholar]
  56. Urcuqui-Inchima S., Haenni A. L., Bernardi F. 2001; Potyvirus proteins: a wealth of functions. Virus Res 74:157–175 [CrossRef]
    [Google Scholar]
  57. Voinnet O. 2005; Induction and suppression of RNA silencing: insights from viral infections. Nat Rev Genet 6:206–220 [CrossRef]
    [Google Scholar]
  58. Voinnet O., Pinto Y. M., Baulcombe D. C. 1999; Suppression of gene silencing: a general strategy used by diverse DNA and RNA viruses of plants. Proc Natl Acad Sci U S A 96:14147–14152 [CrossRef]
    [Google Scholar]
  59. Voinnet O., Rivas S., Mestre P., Baulcombe D. 2003; An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. Plant J 33:949–956 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.82301-0
Loading
/content/journal/jgv/10.1099/vir.0.82301-0
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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