1887

Abstract

Garlic virus X (GarVX) encodes a 15 kDa cysteine-rich protein (CRP). To investigate the function(s) of p15, its subcellular localization, role as a symptom determinant and capacity to act as a viral suppressor of RNA silencing (VSR) were analysed. Results showed that GFP-tagged p15 was distributed in the cytoplasm, nucleus and nucleolus. Expression of p15 from PVX caused additional systemic foliar malformation and led to increased accumulation of PVX, showing that p15 is a virulence factor for reconstructed PVX-p15. Moreover, using a transient agro-infiltration patch assay and a Turnip crinkle virus (TCV) movement complementation assay, it was demonstrated that p15 possesses weak RNA silencing suppressor activity. Removal of an amino acid motif resembling a nuclear localization signal (NLS) prevented p15 from accumulating in the nucleus but did not abolish its silencing suppression activity. This study provides the first insights into the multiple functions of the GarVX p15 protein.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001144
2018-09-12
2019-09-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/99/11/1515.html?itemId=/content/journal/jgv/10.1099/jgv.0.001144&mimeType=html&fmt=ahah

References

  1. Chen J, Zheng HY, Antoniw JF, Adams MJ, Chen JP et al. Detection and classification of allexiviruses from garlic in China. Arch Virol 2004;149:435–445 [CrossRef][PubMed]
    [Google Scholar]
  2. Song SI, Song JT, Kim CH, Lee JS, Choi YD. Molecular characterization of the garlic virus X genome. J Gen Virol 1998;79:155–159 [CrossRef][PubMed]
    [Google Scholar]
  3. Song SI, Song JT, Chang MU, Lee JS, Choi YD. Identification of one of the major viruses infecting garlic plants, garlic virus X. Mol Cells 1997;7:705–709[PubMed]
    [Google Scholar]
  4. Chen J, Chen J, Adams MJ. Molecular characterisation of a complex mixture of viruses in garlic with mosaic symptoms in China. Arch Virol 2001;146:1841–1853 [CrossRef][PubMed]
    [Google Scholar]
  5. Gimenez MD, Yañez-Santos AM, Paz RC, Quiroga MP, Marfil CF et al. Assessment of genetic and epigenetic changes in virus-free garlic (Allium sativum L.) plants obtained by meristem culture followed by in vitro propagation. Plant Cell Rep 2016;35:129–141 [CrossRef][PubMed]
    [Google Scholar]
  6. Wylie SJ, Li H, Saqib M, Jones MG. The global trade in fresh produce and the vagility of plant viruses: a case study in garlic. PLoS One 2014;9:e105044 [CrossRef][PubMed]
    [Google Scholar]
  7. Kang SG, Koo BJ, Lee ET, Chang MU. Allexivirus transmitted by eriophyid mites in garlic plants. J Microbiol Biotechnol 2007;17:1833[PubMed]
    [Google Scholar]
  8. Melo Filho P, Nagata T, Dusi AN, Buso JA, Torres AC et al. Detection of three Allexivirus species infecting garlic in Brazil. Pesquisa Agropecuária Brasileira 2004;39:735–740
    [Google Scholar]
  9. Lu Y, Yan F, Guo W, Zheng H, Lin L et al. Garlic virus X 11-kDa protein granules move within the cytoplasm and traffic a host protein normally found in the nucleolus. Mol Plant Pathol 2011;12:666–676 [CrossRef][PubMed]
    [Google Scholar]
  10. Lu Y, Yin M, Wang X, Chen B, Yang X et al. The unfolded protein response and programmed cell death are induced by expression of Garlic virus X p11 in Nicotiana benthamiana. J Gen Virol 2016;97:1462–1468 [CrossRef][PubMed]
    [Google Scholar]
  11. Adams MJ, Antoniw JF, Bar-Joseph M, Brunt AA, Candresse T et al. The new plant virus family Flexiviridae and assessment of molecular criteria for species demarcation. Arch Virol 2004;149:1672 [CrossRef][PubMed]
    [Google Scholar]
  12. Andika IB, Kondo H, Nishiguchi M, Tamada T. The cysteine-rich proteins of beet necrotic yellow vein virus and tobacco rattle virus contribute to efficient suppression of silencing in roots. J Gen Virol 2012;93:1841–1850 [CrossRef][PubMed]
    [Google Scholar]
  13. Ghazala W, Waltermann A, Pilot R, Winter S, Varrelmann M. Functional characterization and subcellular localization of the 16K cysteine-rich suppressor of gene silencing protein of tobacco rattle virus. J Gen Virol 2008;89:1748–1758 [CrossRef][PubMed]
    [Google Scholar]
  14. Yelina NE et al. Localization of Poa semilatent virus cysteine-rich protein in peroxisomes is dispensable for its ability to suppress RNA silencing. J Gen Virol 2005;86:479–489 [CrossRef]
    [Google Scholar]
  15. Te J, Melcher U, Howard A, Verchot-Lubicz J. Soilborne wheat mosaic virus (SBWMV) 19K protein belongs to a class of cysteine rich proteins that suppress RNA silencing. Virol J 2005;2:18–11 [CrossRef][PubMed]
    [Google Scholar]
  16. Lukhovitskaya NI, Yelina NE, Zamyatnin AA, Schepetilnikov MV, Solovyev AG et al. Expression, localization and effects on virulence of the cysteine-rich 8 kDa protein of Potato mop-top virus. J Gen Virol 2005;86:2879–2889 [CrossRef][PubMed]
    [Google Scholar]
  17. Dunoyer P, Pfeffer S, Fritsch C, Hemmer O, Voinnet O et al. Identification, subcellular localization and some properties of a cysteine-rich suppressor of gene silencing encoded by peanut clump virus. Plant J 2002;29:555–567 [CrossRef][PubMed]
    [Google Scholar]
  18. Fujita N, Komatsu K, Ayukawa Y, Matsuo Y, Hashimoto M et al. N-terminal region of cysteine-rich protein (CRP) in carlaviruses is involved in the determination of symptom types. Mol Plant Pathol 2018;19:180–190 [CrossRef][PubMed]
    [Google Scholar]
  19. Sun L, Andika IB, Kondo H, Chen J. Identification of the amino acid residues and domains in the cysteine-rich protein of Chinese wheat mosaic virus that are important for RNA silencing suppression and subcellular localization. Mol Plant Pathol 2013;14:265–278 [CrossRef][PubMed]
    [Google Scholar]
  20. Liu H, Reavy B, Swanson M, Macfarlane SA. Functional replacement of the tobacco rattle virus cysteine-rich protein by pathogenicity proteins from unrelated plant viruses. Virology 2002;298:232–239 [CrossRef][PubMed]
    [Google Scholar]
  21. Donald RG, Jackson AO. The barley stripe mosaic virus gamma b gene encodes a multifunctional cysteine-rich protein that affects pathogenesis. Plant Cell 1994;6:1593–1606 [CrossRef][PubMed]
    [Google Scholar]
  22. Koonin EV, Boyko VP, Dolja VV. Small cysteine-rich proteins of different groups of plant RNA viruses are related to different families of nucleic acid-binding proteins. Virology 1991;181:395–398 [CrossRef][PubMed]
    [Google Scholar]
  23. Kondo H, Andika IB, Kurokawa E, Bouzoubaa S, Tamada T et al. Comparisons of Rna Silencing Suppressors Encoded by Two Benyviruses, Beet Necrotic Yellow Vein Virus and Burdock Mottle Virus. Symposium of the International Working Group on Plant Viruses with Fungal Vectors 2005
    [Google Scholar]
  24. Carvalho SL, Milanesi DF, Carvalho CM. Functional analysis of Cowpea mild mottle virus triple gene block protein 1 and cysteine-rich protein as potential RNA-silencing suppressors. APS-CPS Joint Meeting 2014
  25. Yelina NE, Erokhina TN, Lukhovitskaya NI, Minina EA, Schepetilnikov MV et al. Localization of Poa semilatent virus cysteine-rich protein in peroxisomes is dispensable for its ability to suppress RNA silencing. J Gen Virol 2005;86:479–489 [CrossRef][PubMed]
    [Google Scholar]
  26. Lukhovitskaya NI, Solovyev AG, Koshkina TE, Zavriev SK, Morozov SY. Interaction of the carlavirus cysteine-rich protein with the plant defense system. Mol Biol 2005;39:785–791 [CrossRef]
    [Google Scholar]
  27. Lukhovitskaya NI, Ignatovich IV, Savenkov EI, Schiemann J, Morozov SY et al. Role of the zinc-finger and basic motifs of chrysanthemum virus B p12 protein in nucleic acid binding, protein localization and induction of a hypersensitive response upon expression from a viral vector. J Gen Virol 2009;90:723–733 [CrossRef][PubMed]
    [Google Scholar]
  28. Deng XG, Peng XJ, Zhu F, Chen YJ, Zhu T et al. A critical domain of sweet potato chlorotic fleck virus nucleotide-binding protein (NaBp) for RNA silencing suppression, nuclear localization and viral pathogenesis. Mol Plant Pathol 2015;16:365–375 [CrossRef][PubMed]
    [Google Scholar]
  29. Li YY, Zhang RN, Xiang HY, Abouelnasr H, Li DW et al. Discovery and characterization of a novel carlavirus infecting potatoes in China. PLoS One 2013;8:e69255 [CrossRef][PubMed]
    [Google Scholar]
  30. Senshu H, Yamaji Y, Minato N, Shiraishi T, Maejima K et al. A dual strategy for the suppression of host antiviral silencing: two distinct suppressors for viral replication and viral movement encoded by potato virus M. J Virol 2011;85:10269–10278 [CrossRef][PubMed]
    [Google Scholar]
  31. Lukhovitskaya NI, Vetukuri RR, Sama I, Thaduri S, Solovyev AG et al. A viral transcription factor exhibits antiviral RNA silencing suppression activity independent of its nuclear localization. J Gen Virol 2014;95:2831–2837 [CrossRef][PubMed]
    [Google Scholar]
  32. Lukhovitskaya NI, Solovieva AD, Boddeti SK, Thaduri S, Solovyev AG et al. An RNA virus-encoded zinc-finger protein acts as a plant transcription factor and induces a regulator of cell size and proliferation in two tobacco species. Plant Cell 2013;25:960–973 [CrossRef][PubMed]
    [Google Scholar]
  33. Lacorte C, Ribeiro SG, Lohuis D, Goldbach R, Prins M. Potatovirus X and tobacco mosaic virus-based vectors compatible with the gateway cloning system. J Virol Methods 2010;164:7–13 [CrossRef][PubMed]
    [Google Scholar]
  34. Huang CH, Hsiao WR, Huang CW, Chen KC, Lin SS et al. Two novel motifs of watermelon silver mottle virus NSs protein are responsible for rna silencing suppression and pathogenicity. PLoS One 2015;10:e0126161 [CrossRef][PubMed]
    [Google Scholar]
  35. Kasschau KD, Xie Z, Allen E, Llave C, Chapman EJ et al. P1/HC-Pro, a viral suppressor of RNA silencing, interferes with Arabidopsis development and miRNA unction. Dev Cell 2003;4:205–217 [CrossRef][PubMed]
    [Google Scholar]
  36. Ding SW, Voinnet O. Antiviral immunity directed by small RNAs. Cell 2007;130:413–426 [CrossRef][PubMed]
    [Google Scholar]
  37. Voinnet O. RNA silencing as a plant immune system against viruses. Trends Genet 2001;17:449–459 [CrossRef][PubMed]
    [Google Scholar]
  38. Voinnet O, Lederer C, Baulcombe DC. A viral movement protein prevents spread of the gene silencing signal in Nicotiana benthamiana. Cell 2000;103:157–167 [CrossRef][PubMed]
    [Google Scholar]
  39. Powers JG, Sit TL, Qu F, Morris TJ, Kim KH et al. A versatile assay for the identification of RNA silencing suppressors based on complementation of viral movement. Mol Plant Microbe Interact 2008;21:879–890 [CrossRef][PubMed]
    [Google Scholar]
  40. Csorba T, Kontra L, Burgyán J. viral silencing suppressors: tools forged to fine-tune host-pathogen coexistence. Virology 2015;479-480:85–103 [CrossRef][PubMed]
    [Google Scholar]
  41. Fernández-Calvino L, Martínez-Priego L, Szabo EZ, Guzmán-Benito I, González I et al. Tobacco rattle virus 16K silencing suppressor binds ARGONAUTE 4 and inhibits formation of RNA silencing complexes. J Gen Virol 2016;97:246–257 [CrossRef][PubMed]
    [Google Scholar]
  42. Murota K, Shimura H, Takeshita M, Masuta C. Interaction between cucumber mosaic virus 2b protein and plant catalase induces a specific necrosis in association with proteasome activity. Plant Cell Rep 2017;36:37–47 [CrossRef][PubMed]
    [Google Scholar]
  43. Desvoyes B, Ramirez-Parra E, Xie Q, Chua NH, Gutierrez C. Cell type-specific role of the retinoblastoma/E2F pathway during Arabidopsis leaf development. Plant Physiol 2006;140:67–80 [CrossRef][PubMed]
    [Google Scholar]
  44. Jiang S, Lu Y, Li K, Lin L, Zheng H et al. Heat shock protein 70 is necessary for Rice stripe virus infection in plants. Mol Plant Pathol 2014;15:907–917 [CrossRef][PubMed]
    [Google Scholar]
  45. Lu Y, McGavin W, Cock PJ, Schnettler E, Yan F et al. Newly identified RNAs of raspberry leaf blotch virus encoding a related group of proteins. J Gen Virol 2015;96:3432–3439 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001144
Loading
/content/journal/jgv/10.1099/jgv.0.001144
Loading

Data & Media loading...

Supplementary File 1

PDF

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