Skip to main content
Log in

HC-Pro hypo- and hypersuppressor mutants: differences in viral siRNA accumulation in vivo and siRNA binding activity in vitro

  • Brief Report
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Viruses have evolved mechanisms to suppress the RNA silencing defense of their hosts, allowing replication and systemic colonization. In a recent study, we found that the effect of mutations in the RNA silencing suppressor of tobacco etch virus (TEV) was variable, ranging from complete abolition of suppressor activity to significantly stronger suppression. Whereas hyposuppressor mutants were less virulent and accumulated fewer viral particles than the wild type, hypersuppressors induced symptoms similar to those of the wild type and accumulated particles to similar levels. Here, we further characterize a set of these mutants in terms of their ability to bind in vitro and induce accumulation in vivo of virus-derived siRNAs. Hyposuppressor alleles are less efficient at binding siRNAs than hypersuppressors, whereas the latter are not different from the wild type. As a consequence of lower viral accumulation, plants infected with virus bearing a hyposuppressor allele also accumulate less virus-derived siRNA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

References

  1. Anandalakshmi R, Pruss GJ, Ge X, Marathe R, Mallory AC, Smith TH, Vance VB (1998) A viral suppressor of gene silencing in plants. Proc Natl Acad Sci USA 95:13079–13084

    Article  CAS  PubMed  Google Scholar 

  2. Anandalakshmi R, Marathe R, Ge X, Herr JM, Mau C, Mallory A, Pruss G, Bowman L, Vance VB (2000) A calmodulin-related protein that suppresses posttranscriptional gene silencing in plants. Science 290:142–144

    Article  CAS  PubMed  Google Scholar 

  3. Brigneti G, Voinnet O, Li WX, Ji LH, Ding SW, Baulcombe DC (1998) Viral pathogenicity determinants are suppressors of transgene silencing in Nicotiana benthamiana. EMBO J 17:6739–6746

    Article  CAS  PubMed  Google Scholar 

  4. Deleris A, Gallego-Bartolome J, Bao JS, Kasschau KD, Carrington JC, Voinnet O (2006) Hierarchical action and inhibition of plant Dicer-like proteins in antiviral defense. Science 313:68–71

    Article  CAS  PubMed  Google Scholar 

  5. Dunoyer P, Voinnet O (2008) Mixing and matching: the essence of plant systemic silencing? Trends Genet 24:151–154

    Article  CAS  PubMed  Google Scholar 

  6. Ebhardt HA, Thi EP, Wang MB, Unrau PJ (2005) Extensive 3′ modification of plant small RNAs is modulated by helper component-proteinase expression. Proc Natl Acad Sci USA 102:13398–13403

    Article  CAS  PubMed  Google Scholar 

  7. Johansen LK, Carrington JC (2001) Silencing on the spot: induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system. Plant Physiol 126:930–938

    Article  CAS  PubMed  Google Scholar 

  8. Kasschau KD, Carrington JC (1998) A counterdefensive strategy of plant viruses: suppression of posttranscriptional gene silencing. Cell 95:461–470

    Article  CAS  PubMed  Google Scholar 

  9. Lakatos L, Csorba T, Pantaleo V, Chapman EJ, Carrington JC, Liu YP, Dolja VV, Fernández Calvino L, López-Moya JJ, Burgyán J (2006) Small RNA binding is a common strategy to suppress RNA silencing by several viral suppressors. EMBO J 25:2768–2780

    Article  CAS  PubMed  Google Scholar 

  10. Lozsa R, Csorba T, Lakatos L, Burgyán J (2008) Inhibition of 3′ modification of small RNAs in virus-infected plants require spatial and temporal co-expression of small RNAs and viral silencing-suppressor proteins. Nucleic Acids Res 36:4099–4107

    Article  CAS  PubMed  Google Scholar 

  11. Mérai Z, Kerényi Z, Kertész S, Magna M, Lakatos L, Silhavy D (2006) Double-stranded RNA binding may be a general plant RNA viral strategy to suppress RNA silencing. J Virol 80:5747–5756

    Article  PubMed  Google Scholar 

  12. Mérai Z, Kerényi Z, Molnár A, Barta E, Válóczi A, Bisztray G, Havelda Z, Burgyán J, Silhavy D (2005) Aureusvirus P14 is an efficient RNA silencing suppressor that binds double-stranded RNAs without size specificity. J Virol 79:7217–7226

    Article  PubMed  Google Scholar 

  13. Plisson C, Druker 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

    Article  CAS  PubMed  Google Scholar 

  14. Pruss G, Ge X, Shi XM, Carrington JC, Vance VB (1997) Plant viral synergism: the potyviral genome encodes a broad-range pathogenicity enhancer that transactivates replication of heterologous viruses. Plant Cell 9:859–868

    Article  CAS  PubMed  Google Scholar 

  15. Sokal RR, Rohlf FJ (1995) Biometry. Freeman and Co, New York

    Google Scholar 

  16. Torres-Barceló C, Martín S, Daròs JA, Elena SF (2008) From Hypo- to Hypersuppression: effect of amino acid substitutions on the RNA-silencing suppressor activity of the Tobacco etch potyvirus HC-Pro. Genetics 180:1039–1049

    Article  PubMed  Google Scholar 

  17. Varrelman M, Maiss E, Pilot R, Palkovics L (2007) Use of pentapeptide-insertion scanning mutagenesis for functional mapping of the Plum pox virus helper-component proteinase suppressor of gene silencing. J Gen Virol 88:1005–1015

    Article  Google Scholar 

  18. Yu B, Chapman EJ, Yang ZY, Carrington JC, Chen XM (2006) Transgenically expressed viral RNA silencing suppressors interfere with microRNA methylation in Arabidopsis. FEBS Lett 580:3117–3120

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank J. Marqués, S. Martín and M.Á. Nohales for comments and suggestions, Z. Mérai and A. Valli for protocols, and A. Cuadrado and F. de la Iglesia for excellent technical assistance. This work was supported by grants from the Spanish Ministerio de Ciencia e Innovación (MICINN)-Fondo Europeo de Desarrollo Regional (BFU2006-14819-C02-01/BMC), the Generalitat Valenciana (ACOMP07-263) and the European Molecular Biology Organization Young Investigator Program to S.F.E and by grant BIO2008-01986 (MICINN) to J.A.D. C.T.B. was supported by a predoctoral fellowship from the Generalitat Valenciana.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Santiago F. Elena.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Torres-Barceló, C., Daròs, JA. & Elena, S.F. HC-Pro hypo- and hypersuppressor mutants: differences in viral siRNA accumulation in vivo and siRNA binding activity in vitro. Arch Virol 155, 251–254 (2010). https://doi.org/10.1007/s00705-009-0563-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-009-0563-x

Keywords

Navigation