1887

Abstract

Mutations within the HC-Pro coding region of (WSMV) were introduced by misincorporation during PCR and evaluated for phenotype within the context of an infectious clone. Nine synonymous substitutions and 15 of 25 non-synonymous substitutions had no phenotypic effect. Four non-synonymous substitutions, including one that reverted consistently to wild type, resulted in attenuated systemic infection. Six non-synonymous substitutions and one nonsense substitution abolished systemic infectivity. Mutants bearing the GUS reporter gene were evaluated for the ability to establish primary infection foci. All attenuated mutants and two systemic infection-deficient mutants produced localized regions of GUS expression on inoculated leaves 3 days post-inoculation. assays revealed that mutants able to establish infection foci retained HC-Pro proteinase activity. Among mutants unable to establish infection foci, HC-Pro proteinase activity was retained, reduced or absent. As a complete HC-Pro deletion mutant can infect plants systemically, certain substitutions in this dispensable gene probably prevented infection of WSMV via interference.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.81933-0
2006-09-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/87/9/2741.html?itemId=/content/journal/jgv/10.1099/vir.0.81933-0&mimeType=html&fmt=ahah

References

  1. Ammar E. D., Järlfors U., Pirone T. P. 1994; Association of potyvirus helper component protein with virions and the cuticle lining the maxillary food canal and foregut of an aphid vector. Phytopathology 84:1054–1060 [CrossRef]
    [Google Scholar]
  2. Atreya C. D., Pirone T. P. 1993; Mutational analysis of the helper component-proteinase gene of a potyvirus: effects of amino acid substitutions, deletions, and gene replacement on virulence and aphid transmissibility. Proc Natl Acad Sci U S A 90:11919–11923 [CrossRef]
    [Google Scholar]
  3. Atreya C. D., Atreya P. L., Thornbury D. W., Pirone T. P. 1992; Site-directed mutations in the potyvirus HC-PRO gene affect helper component activity, virus accumulation, and symptom expression in infected tobacco plants. Virology 191:106–111 [CrossRef]
    [Google Scholar]
  4. Berger P. H., Pirone T. P. 1986; The effect of helper component on the uptake and localization of potyviruses in Myzus persicae . Virology 153:256–261 [CrossRef]
    [Google Scholar]
  5. Blanc S., López-Moya J.-J., Wang R. Y., García-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]
  6. 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]
  7. Carrington J. C., Herndon K. L. 1992; Characterization of the potyviral HC-Pro autoproteolytic cleavage site. Virology 187:308–315 [CrossRef]
    [Google Scholar]
  8. 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]
  9. Choi I.-R., French R., Hein G. L., Stenger D. C. 1999; Fully biologically active in vitro transcripts of the eriophyid mite-transmitted wheat streak mosaic tritimovirus. Phytopathology 89:1182–1185 [CrossRef]
    [Google Scholar]
  10. Choi I.-R., Stenger D. C., French R. 2000a; Multiple interactions among proteins encoded by the mite-transmitted wheat streak mosaic tritimovirus. Virology 267:185–198 [CrossRef]
    [Google Scholar]
  11. Choi I.-R., Stenger D. C., Morris T. J., French R. 2000b; A plant virus vector for systemic expression of foreign genes in cereals. Plant J 23:547–555 [CrossRef]
    [Google Scholar]
  12. Choi I.-R., Horken K. M., Stenger D. C., French R. 2002; Mapping of the P1 proteinase cleavage site in the polyprotein of Wheat streak mosaic virus (genus Tritimovirus . J Gen Virol 83:443–450
    [Google Scholar]
  13. Choi I.-R., Horken K. M., Stenger D. C., French R. 2005; An internal RNA element in the P3 cistron of Wheat streak mosaic virus revealed by synonymous mutations that affect both movement and replication. J Gen Virol 86:2605–2614 [CrossRef]
    [Google Scholar]
  14. 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]
  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. French R., Stenger D. C. 2005; Genome sequences of Agropyron mosaic virus and Hordeum mosaic virus support reciprocal monophyly of the genera Potyvirus and Rymovirus in the family Potyviridae . Arch Virol 150:299–312 [CrossRef]
    [Google Scholar]
  17. Govier D. A., Kassanis B. 1974; A virus-induced component of plant sap needed when aphids acquire potato virus Y from purified preparations. Virology 61:420–426 [CrossRef]
    [Google Scholar]
  18. Hall J. S., French R., Morris T. J., Stenger D. C. 2001; Structure and temporal dynamics of populations within wheat streak mosaic virus isolates. J Virol 75:10231–10243 [CrossRef]
    [Google Scholar]
  19. Huet H., Gal-On A., Meir E., Lecoq H., Raccah B. 1994; Mutations in the helper component protease gene of zucchini yellow mosaic virus affect its ability to mediate aphid transmissibility. J Gen Virol 75:1407–1414 [CrossRef]
    [Google Scholar]
  20. 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]
  21. Kasschau K. D., Carrington J. C. 2001; Long-distance movement and replication maintenance functions correlate with silencing suppression activity of potyviral HC-Pro. Virology 285:71–81 [CrossRef]
    [Google Scholar]
  22. 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]
  23. McNeil J. E., French R., Hein G. L., Baenziger P. S., Eskridge K. M. 1996; Characterization of genetic variability among natural populations of wheat streak mosaic virus. Phytopathology 86:1222–1227 [CrossRef]
    [Google Scholar]
  24. Oh C.-S., Carrington J. C. 1989; Identification of essential residues in potyvirus proteinase HC-Pro by site-directed mutagenesis. Virology 173:692–699 [CrossRef]
    [Google Scholar]
  25. Peng Y., 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]
  26. Stenger D. C., French R. 2004; Functional replacement of Wheat streak mosaic virus HC-Pro with the corresponding cistron from a diverse array of viruses in the family Potyviridae . Virology 323:257–267 [CrossRef]
    [Google Scholar]
  27. Stenger D. C., Hall J. S., Choi I.-R., French R. 1998; Phylogenetic relationships within the family Potyviridae : wheat streak mosaic virus and brome streak mosaic virus are not members of the genus Rymovirus . Phytopathology 88:782–787 [CrossRef]
    [Google Scholar]
  28. Stenger D. C., French R., Gildow F. E. 2005a; Complete deletion of Wheat streak mosaic virus HC-Pro: a null mutant is viable for systemic infection. J Virol 79:12077–12080 [CrossRef]
    [Google Scholar]
  29. Stenger D. C., Hein G. L., Gildow F. E., Horken K. M., French R. 2005b; Plant virus HC-Pro is a determinant of eriophyid mite transmission. J Virol 79:9054–9061 [CrossRef]
    [Google Scholar]
  30. Stenger D. C., Hein G. L., French R. 2006; Nested deletion analysis of Wheat streak mosaic virus HC-Pro: mapping of domains affecting polyprotein processing and eriophyid mite transmission. Virology 350:465–474 [CrossRef]
    [Google Scholar]
  31. Thornbury D. W., Hellmann G. M., Rhoads R. E., Pirone T. P. 1985; Purification and characterization of potyvirus helper component. Virology 144:260–267 [CrossRef]
    [Google Scholar]
  32. Wang R. Y., Ammar E. D., Thornbury D. W., Lopez-Moya J. J., Pirone T. P. 1996; Loss of potyvirus transmissibility and helper-component activity correlate with non-retention of virions in aphid stylets. J Gen Virol 77:861–867 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.81933-0
Loading
/content/journal/jgv/10.1099/vir.0.81933-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
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