1887

Abstract

In plants, RNA silencing functions as a potent antiviral mechanism. Virus-derived double-stranded RNAs (dsRNAs) trigger this mechanism, being cleaved by Dicer-like (DCL) enzymes into virus small RNAs (vsRNAs). These vsRNAs guide sequence-specific RNA degradation upon their incorporation into an RNA-induced silencing complex (RISC) that contains a slicer of the (AGO) family. Host RNA dependent-RNA polymerases, particularly RDR6, strengthen antiviral silencing by generating more dsRNA templates from RISC-cleavage products that, in turn, are converted into secondary vsRNAs by DCLs. Previous work showed that (PLPV) is a very efficient inducer and target of RNA silencing as PLPV-infected plants accumulate extraordinarily high amounts of vsRNAs that, strikingly, are independent of RDR6 activity. Several scenarios may explain these observations including a major contribution of dicing versus slicing for defence against PLPV, as the dicing step would not be affected by the RNA silencing suppressor encoded by the virus, a protein that acts via vsRNA sequestration. Taking advantage of the availability of lines of with DCL or AGO2 functions impaired, here we have tried to get further insights into the components of the silencing machinery that are involved in anti-PLPV-silencing. Results have shown that DCL4 and, to lesser extent, DCL2 contribute to restrict viral infection. Interestingly, AGO2 apparently makes even a higher contribution in the defence against PLPV, extending the number of viruses that are affected by this particular slicer. The data support that both dicing and slicing activities participate in the host race against PLPV.

Funding
This study was supported by the:
  • general secretary for research and technology of greece (Award AGRO4CRETE - 2018ΣΕ01300000)
    • Principle Award Recipient: KonstantinaKatsarou
  • conselleria d'educació, investigació, cultura i esport (Award PROMETEO/2019/012)
    • Principle Award Recipient: CarmenHernández
  • ministerio de economía y competitividad (Award BFU2015-70261)
    • Principle Award Recipient: CarmenHernández
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001656
2021-10-08
2021-10-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/102/10/jgv001656.html?itemId=/content/journal/jgv/10.1099/jgv.0.001656&mimeType=html&fmt=ahah

References

  1. Csorba T, Pantaleo V, Burgyán J. RNA silencing: an antiviral mechanism. Adv Virus Res 2009; 75:35–71 [View Article] [PubMed]
    [Google Scholar]
  2. Ding S-W. RNA-based antiviral immunity. Nat Rev Immunol 2010; 10:632–644 [View Article] [PubMed]
    [Google Scholar]
  3. Ding S-W, Voinnet O. Antiviral immunity directed by small RNAs. Cell 2007; 130:413–426 [View Article] [PubMed]
    [Google Scholar]
  4. Llave C. Virus-derived small interfering RNAs at the core of plant-virus interactions. Trends Plant Sci 2010; 15:701–707 [View Article] [PubMed]
    [Google Scholar]
  5. Mallory A, Vaucheret H. Form, function, and regulation of ARGONAUTE proteins. Plant Cell 2010; 22:3879–3889 [View Article] [PubMed]
    [Google Scholar]
  6. Silva-Martins G, Bolaji A, Moffett P. What does it take to be antiviral? An Argonaute-centered perspective on plant antiviral defense. J Exp Bot 2020; 71:6197–6210 [View Article] [PubMed]
    [Google Scholar]
  7. Baulcombe D. RNA silencing in plants. Nature 2004; 431:356–363 [View Article] [PubMed]
    [Google Scholar]
  8. Wassenegger M, Krczal G. Nomenclature and functions of RNA-directed RNA polymerases. Trends Plant Sci 2006; 11:142–151 [View Article] [PubMed]
    [Google Scholar]
  9. Pyott DE, Molnar A. Going mobile: non-cell-autonomous small RNAs shape the genetic landscape of plants. Plant Biotechnol J 2015; 13:306–318 [View Article] [PubMed]
    [Google Scholar]
  10. Blevins T, Rajeswaran R, Shivaprasad PV, Beknazariants D, Si-Ammour A. Four plant Dicers mediate viral small RNA biogenesis and DNA virus induced silencing. Nucleic Acids Res 2006; 34:6233–6246 [View Article] [PubMed]
    [Google Scholar]
  11. Deleris A, Gallego-Bartolome J, Bao J, Kasschau KD, Carrington JC. Hierarchical action and inhibition of plant Dicer-like proteins in antiviral defense. Science 2006; 313:68–71 [View Article] [PubMed]
    [Google Scholar]
  12. Liu Q, Feng Y, Zhu Z. Dicer-like (DCL) proteins in plants. Funct Integr Genomics 2009; 9:277–286 [View Article] [PubMed]
    [Google Scholar]
  13. Diaz-Pendon JA, Li F, Li W-X, Ding S-W. Suppression of antiviral silencing by cucumber mosaic virus 2b protein in arabidopsis is associated with drastically reduced accumulation of three classes of viral small interfering rnas. Plant Cell 2007; 19:2053–2063 [View Article] [PubMed]
    [Google Scholar]
  14. Harvey JJW, Lewsey MG, Patel K, Westwood J, Heimstädt S. An antiviral defense role of AGO2 in plants. PLoS One 2011; 6:e14639 [View Article] [PubMed]
    [Google Scholar]
  15. Morel J-B, Godon C, Mourrain P, Béclin C, Boutet S et al. Fertile hypomorphic ARGONAUTE (ago1) mutants impaired in post-transcriptional gene silencing and virus resistance. Plant Cell 2002; 14:629–639 [View Article] [PubMed]
    [Google Scholar]
  16. Qu F, Ye X, Morris TJ. Arabidopsis DRB4, AGO1, AGO7, and RDR6 participate in a DCL4-initiated antiviral RNA silencing pathway negatively regulated by DCL1. Proc Natl Acad Sci U S A 2008; 105:14732–14737 [View Article] [PubMed]
    [Google Scholar]
  17. Pumplin N, Voinnet O. RNA silencing suppression by plant pathogens: defence, counter-defence and counter-counter-defence. Nat Rev Microbiol 2013; 11:745–760 [View Article] [PubMed]
    [Google Scholar]
  18. Lakatos L, Csorba T, Pantaleo V, Chapman EJ, Carrington JC. Small RNA binding is a common strategy to suppress RNA silencing by several viral suppressors. EMBO J 2006; 25:2768–2780 [View Article] [PubMed]
    [Google Scholar]
  19. Mérai Z, Kerényi Z, Kertész S, Magna M, Lakatos L. Double-stranded RNA binding may be a general plant RNA viral strategy to suppress RNA silencing. J Virol 2006; 80:5747–5756 [View Article] [PubMed]
    [Google Scholar]
  20. Vargason JM, Szittya G, Burgyán J, Hall TMT. Size selective recognition of siRNA by an RNA silencing suppressor. Cell 2003; 115:799–811 [View Article] [PubMed]
    [Google Scholar]
  21. Ye K, Malinina L, Patel DJ. Recognition of small interfering RNA by a viral suppressor of RNA silencing. Nature 2003; 426:874–878 [View Article] [PubMed]
    [Google Scholar]
  22. Csorba T, Kontra L, Burgyán J. Viral silencing suppressors: tools forged to fine-tune host-pathogen coexistence. Virology 2015; 479–480:85–103 [View Article] [PubMed]
    [Google Scholar]
  23. Flynt A, Liu N, Martin R, Lai EC. Dicing of viral replication intermediates during silencing of latent Drosophila viruses. Proc Natl Acad Sci U S A 2009; 106:5270–5275 [View Article] [PubMed]
    [Google Scholar]
  24. Omarov RT, Ciomperlik JJ, Scholthof HB. RNAi-associated ssRNA-specific ribonucleases in Tombusvirus P19 mutant-infected plants and evidence for a discrete siRNA-containing effector complex. Proc Natl Acad Sci U S A 2007; 104:1714–1719 [View Article] [PubMed]
    [Google Scholar]
  25. Pantaleo V, Szittya G, Burgyán J. Molecular bases of viral RNA targeting by viral small interfering RNA-programmed RISC. J Virol 2007; 81:3797–3806 [View Article] [PubMed]
    [Google Scholar]
  26. Takahashi H, Fukuhara T, Kitazawa H, Kormelink R. Virus latency and the impact on plants. Front Microbiol 2019; 10:2764 [View Article] [PubMed]
    [Google Scholar]
  27. Roossinck MJ. Lifestyles of plant viruses. Philos Trans R Soc Lond B Biol Sci 2010; 365:1899–1905 [View Article] [PubMed]
    [Google Scholar]
  28. Scheets K, Jordan R, White KA, Hernández C. Pelarspovirus, a proposed new genus in the family Tombusviridae. Arch Virol 2015; 160:2385–2393 [View Article] [PubMed]
    [Google Scholar]
  29. Castaño A, Ruiz L, Hernández C. Insights into the translational regulation of biologically active open reading frames of Pelargonium line pattern virus. Virology 2009; 386:417–426 [View Article] [PubMed]
    [Google Scholar]
  30. Castaño A, Hernández C. Complete nucleotide sequence and genome organization of Pelargonium line pattern virus and its relationship with the family Tombusviridae. Arch Virol 2005; 150:949–965 [View Article] [PubMed]
    [Google Scholar]
  31. Pérez-Cañamás M, Hernández C. Key importance of small RNA binding for the activity of a glycine-tryptophan (GW) motif-containing viral suppressor of RNA silencing. J Biol Chem 2015; 290:3106–3120 [View Article] [PubMed]
    [Google Scholar]
  32. Pérez-Cañamás M, Blanco-Pérez M, Forment J, Hernández C. Nicotiana benthamiana plants asymptomatically infected by Pelargonium line pattern virus show unusually high accumulation of viral small RNAs that is neither associated with DCL induction nor RDR6 activity. Virology 2017; 501:136–146 [View Article] [PubMed]
    [Google Scholar]
  33. Dadami E, Boutla A, Vrettos N, Tzortzakaki S, Karakasilioti I. DICER-LIKE 4 but not DICER-LIKE 2 may have a positive effect on potato spindle tuber viroid accumulation in Nicotiana benthamiana. Mol Plant 2013; 6:232–234 [View Article] [PubMed]
    [Google Scholar]
  34. Katsarou K, Mavrothalassiti E, Dermauw W, Van Leeuwen T, Kalantidis K. Combined activity of DCL2 and DCL3 is crucial in the defense against potato spindle tuber viroid. PLoS Pathog 2016; 12:e1005936 [View Article] [PubMed]
    [Google Scholar]
  35. Ludman M, Burgyán J, Fátyol K. Crispr/Cas9 mediated inactivation of argonaute 2 reveals its differential involvement in antiviral responses. Sci Rep 2017; 7:1010 [View Article] [PubMed]
    [Google Scholar]
  36. Verwoerd TC, Dekker BMM, Hoekema A. A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 1989; 17:2362 [View Article] [PubMed]
    [Google Scholar]
  37. Nakasugi K, Crowhurst RN, Bally J, Wood CC, Hellens RP. De novo transcriptome sequence assembly and analysis of RNA silencing genes of Nicotiana benthamiana. PLoS One 2013; 8:e59534 [View Article] [PubMed]
    [Google Scholar]
  38. Donaire L, Barajas D, Martínez-García B, Martínez-Priego L, Pagán I. Structural and genetic requirements for the biogenesis of tobacco rattle virus-derived small interfering RNAs. J Virol 2008; 82:5167–5177 [View Article] [PubMed]
    [Google Scholar]
  39. Garcia-Ruiz H, Takeda A, Chapman EJ, Sullivan CM, Fahlgren N. Arabidopsis RNA-dependent RNA polymerases and dicer-like proteins in antiviral defense and small interfering RNA biogenesis during Turnip Mosaic Virus infection. Plant Cell 2010; 22:481–496 [View Article] [PubMed]
    [Google Scholar]
  40. Katsarou K, Mitta E, Bardani E, Oulas A, Dadami E. DCL-suppressed Nicotiana benthamiana plants: valuable tools in research and biotechnology. Mol Plant Pathol 2019; 20:432–446 [View Article] [PubMed]
    [Google Scholar]
  41. Jaubert M, Bhattacharjee S, Mello AFS, Perry KL, Moffett P. ARGONAUTE2 mediates RNA-silencing antiviral defenses against Potato virus X in Arabidopsis. Plant Physiol 2011; 156:1556–1564 [View Article] [PubMed]
    [Google Scholar]
  42. Scholthof HB, Alvarado VY, Vega-Arreguin JC, Ciomperlik J, Odokonyero D. Identification of an ARGONAUTE for antiviral RNA silencing in Nicotiana benthamiana. Plant Physiol 2011; 156:1548–1555 [View Article] [PubMed]
    [Google Scholar]
  43. Odokonyero D, Mendoza MR, Alvarado VY, Zhang J, Wang X. Transgenic down-regulation of ARGONAUTE2 expression in Nicotiana benthamiana interferes with several layers of antiviral defenses. Virology 2015; 486:209–218 [View Article] [PubMed]
    [Google Scholar]
  44. Odokonyero D, Mendoza MR, Moffett P, Scholthof HB. Tobacco rattle virus (TRV)-Mediated Silencing of Nicotiana benthamiana ARGONAUTES (NbAGOs) reveals new antiviral candidates and dominant effects of TRV-NbAGO1. Phytopathology 2017; 107:977–987 [View Article] [PubMed]
    [Google Scholar]
  45. Diao P, Zhang Q, Sun H, Ma W, Cao A et al. miR403a and SA Are Involved in NbAGO2 mediated antiviral defenses against TMV infection in Nicotiana benthamiana. Genes (Basel) 2019; 10: [View Article] [PubMed]
    [Google Scholar]
  46. Wang X-B, Jovel J, Udomporn P, Wang Y, Wu Q et al. The 21-nucleotide, but not 22-nucleotide, viral secondary small interfering RNAs direct potent antiviral defense by two cooperative argonautes in Arabidopsis thaliana. Plant Cell 2011; 23:1625–1638 [View Article] [PubMed]
    [Google Scholar]
  47. Carbonell A, Fahlgren N, Garcia-Ruiz H, Gilbert KB, Montgomery TA. Functional analysis of three Arabidopsis ARGONAUTES using slicer-defective mutants. Plant Cell 2012; 24:3613–3629 [View Article] [PubMed]
    [Google Scholar]
  48. Garcia-Ruiz H, Carbonell A, Hoyer JS, Fahlgren N, Gilbert KB. Roles and programming of Arabidopsis ARGONAUTE proteins during Turnip mosaic virus infection. PLoS Pathog 2015; 11:e1004755 [View Article] [PubMed]
    [Google Scholar]
  49. Ma X, Nicole M-C, Meteignier L-V, Hong N, Wang G et al. Different roles for RNA silencing and RNA processing components in virus recovery and virus-induced gene silencing in plants. J Exp Bot 2015; 66:919–932 [View Article] [PubMed]
    [Google Scholar]
  50. Zhang X, Zhang X, Singh J, Li D, Qu F. Temperature-dependent survival of Turnip crinkle virus-infected arabidopsis plants relies on an RNA silencing-based defense that requires dcl2, AGO2, and HEN1. J Virol 2012; 86:6847–6854 [View Article] [PubMed]
    [Google Scholar]
  51. Ghoshal B, Sanfaçon H. Temperature-dependent symptom recovery in Nicotiana benthamiana plants infected with tomato ringspot virus is associated with reduced translation of viral RNA2 and requires ARGONAUTE 1. Virology 2014; 456–457:188–197 [View Article] [PubMed]
    [Google Scholar]
  52. Paudel DB, Ghoshal B, Jossey S, Ludman M, Fatyol K. Expression and antiviral function of ARGONAUTE 2 in Nicotiana benthamiana plants infected with two isolates of tomato ringspot virus with varying degrees of virulence. Virology 2018; 524:127–139 [View Article] [PubMed]
    [Google Scholar]
  53. Mi S, Cai T, Hu Y, Chen Y, Hodges E et al. Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5’ terminal nucleotide. Cell 2008; 133:116–127 [View Article] [PubMed]
    [Google Scholar]
  54. Takeda A, Iwasaki S, Watanabe T, Utsumi M, Watanabe Y. The mechanism selecting the guide strand from small RNA duplexes is different among argonaute proteins. Plant Cell Physiol 2008; 49:493–500 [View Article] [PubMed]
    [Google Scholar]
  55. Ludman M, Fátyol K. Targeted inactivation of the AGO1 homeologues of Nicotiana benthamiana reveals their distinct roles in development and antiviral defence. New Phytol 2021; 229:1289–1297 [View Article] [PubMed]
    [Google Scholar]
  56. Schauer SE, Jacobsen SE, Meinke DW, Ray A. DICER-LIKE1: blind men and elephants in Arabidopsis development. Trends Plant Sci 2002; 7:487–491 [View Article] [PubMed]
    [Google Scholar]
  57. Gutiérrez S, Yvon M, Pirolles E, Garzo E, Fereres A. Circulating virus load determines the size of bottlenecks in viral populations progressing within a host. PLoS Pathog 2012; 8:e1003009 [View Article] [PubMed]
    [Google Scholar]
  58. Cordero T, Cerdán L, Carbonell A, Katsarou K, Kalantidis K. Dicer-Like 4 is involved in restricting the systemic movement of Zucchini yellow mosaic virus in Nicotiana benthamiana. Mol Plant Microbe Interact 2017; 30:63–71 [View Article] [PubMed]
    [Google Scholar]
  59. Pérez-Cañamás M, Hernández C. New insights into the nucleolar localization of a plant RNA virus-encoded protein that acts in both RNA packaging and RNA silencing suppression: involvement of importins alpha and relevance for viral infection. Mol Plant Microbe Interact 2018; 31:1134–1144 [View Article] [PubMed]
    [Google Scholar]
  60. Kim SH, Ryabov EV, Kalinina NO, Rakitina DV, Gillespie T. Cajal bodies and the nucleolus are required for a plant virus systemic infection. EMBO J 2007; 26:2169–2179 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001656
Loading
/content/journal/jgv/10.1099/jgv.0.001656
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