1887

Abstract

The Lolium latent virus (LoLV) major coat protein sequence contains a typical chloroplast transit peptide (cTP) domain. In infected Nicotiana benthamiana leaf tissue, LoLV coat proteins can be detected at the chloroplast. In transient expression, several N-terminal deletions of the CP sequence, increasing in length, result in disruption of the domain functionality, markedly affecting intracellular localization. A yeast two-hybrid-based study using LoLV CP as bait identified several potentially interacting Arabidopsis host proteins, most of them with chloroplast-linked pathways. One of them, an ankyrin repeat protein, was studied in detail. The N. benthamiana homologue (NbANKr) targets chloroplasts, is able to co-localize with LoLV CP at chloroplast membranes in transient expression and shows a robust interaction with LoLV CP in vivo by BiFC, which has been confirmed by yeast two-hybrid data. Silencing NbANKr genes in N. benthamiana plants, prior to challenging with LoLV by mechanical inoculation, affects LoLV infection, significantly reducing the level of viral RNA in young leaves, compared to levels in control plants, and suggesting an inhibition of virus movement. Silencing of NbANKr has no obvious effect on plant phenotype, but is able to interfere with LoLV infection, opening the way for a new strategy for virus infection control.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001043
2018-03-20
2019-09-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/99/5/730.html?itemId=/content/journal/jgv/10.1099/jgv.0.001043&mimeType=html&fmt=ahah

References

  1. Vaira AM, Lim HS, Bauchan GR, Owens RA, Natilla A et al. Lolium latent virus (Alphaflexiviridae) coat proteins: expression and functions in infected plant tissue. J Gen Virol 2012; 93: 1814– 1824 [CrossRef] [PubMed]
    [Google Scholar]
  2. den Boon JA, Ahlquist P. Organelle-like membrane compartmentalization of positive-strand RNA virus replication factories. Annu Rev Microbiol 2010; 64: 241– 256 [CrossRef] [PubMed]
    [Google Scholar]
  3. Novoa RR, Calderita G, Arranz R, Fontana J, Granzow H et al. Virus factories: associations of cell organelles for viral replication and morphogenesis. Biol Cell 2005; 97: 147– 172 [CrossRef] [PubMed]
    [Google Scholar]
  4. Hyodo K, Okuno T. Host factors used by positive-strand RNA plant viruses for genome replication. J Gen Plant Pathol 2014; 80: 123– 135 [CrossRef]
    [Google Scholar]
  5. Padmanabhan MS, Dinesh-Kumar SP. All hands on deck—the role of chloroplasts, endoplasmic reticulum, and the nucleus in driving plant innate immunity. Mol Plant Microbe Interact 2010; 23: 1368– 1380 [CrossRef] [PubMed]
    [Google Scholar]
  6. Jang C, Seo EY, Nam J, Bae H, Gim YG et al. Insights into alternanthera mosaic virus TGB3 functions: interactions with Nicotiana benthamiana PsbO correlate with chloroplast vesiculation and veinal necrosis caused by TGB3 over-expression. Front Plant Sci 2013; 4: 5 [CrossRef] [PubMed]
    [Google Scholar]
  7. Zhao J, Zhang X, Hong Y, Liu Y. Chloroplast in plant-virus interaction. Front Microbiol 2016; 7: 1565 [CrossRef] [PubMed]
    [Google Scholar]
  8. Breeden L, Nasmyth K. Similarity between cell-cycle genes of budding yeast and fission yeast and the Notch gene of Drosophila. Nature 1987; 329: 651– 654 [CrossRef] [PubMed]
    [Google Scholar]
  9. Artavanis-Tsakonas S, Delidakis C, Fehon RG. The Notch locus and the cell biology of neuroblast segregation. Annu Rev Cell Biol 1991; 7: 427– 452 [CrossRef] [PubMed]
    [Google Scholar]
  10. Bennett V. Ankyrins. Adaptors between diverse plasma membrane proteins and the cytoplasm. J Biol Chem 1992; 267: 8703– 8706 [PubMed]
    [Google Scholar]
  11. Sedgwick SG, Smerdon SJ. The ankyrin repeat: a diversity of interactions on a common structural framework. Trends Biochem Sci 1999; 24: 311– 316 [CrossRef] [PubMed]
    [Google Scholar]
  12. Bae W, Lee YJ, Kim DH, Lee J, Kim S et al. AKR2A-mediated import of chloroplast outer membrane proteins is essential for chloroplast biogenesis. Nat Cell Biol 2008; 10: 220– 227 [CrossRef] [PubMed]
    [Google Scholar]
  13. Cui YL, Jia QS, Yin QQ, Lin GN, Kong MM et al. The GDC1 gene encodes a novel ankyrin domain-containing protein that is essential for grana formation in Arabidopsis. Plant Physiol 2011; 155: 130– 141 [CrossRef] [PubMed]
    [Google Scholar]
  14. Zhang H, Li X, Zhang Y, Kuppu S, Shen G. Is AKR2A an essential molecular chaperone for a class of membrane-bound proteins in plants?. Plant Signal Behav 2010; 5: 1520– 1522 [CrossRef] [PubMed]
    [Google Scholar]
  15. Lim HS, Bragg JN, Ganesan U, Ruzin S, Schichnes D et al. Subcellular localization of the barley stripe mosaic virus triple gene block proteins. J Virol 2009; 83: 9432– 9448 [CrossRef] [PubMed]
    [Google Scholar]
  16. Jackson DT, Froehlich JE, Keegstra K. The hydrophilic domain of Tic110, an inner envelope membrane component of the chloroplastic protein translocation apparatus, faces the stromal compartment. J Biol Chem 1998; 273: 16583– 16588 [CrossRef] [PubMed]
    [Google Scholar]
  17. Xiang Y, Kakani K, Reade R, Hui E, Rochon D. A 38-amino-acid sequence encompassing the arm domain of the cucumber necrosis virus coat protein functions as a chloroplast transit peptide in infected plants. J Virol 2006; 80: 7952– 7964 [CrossRef] [PubMed]
    [Google Scholar]
  18. Yan J, Wang J, Zhang H. An ankyrin repeat-containing protein plays a role in both disease resistance and antioxidation metabolism. Plant J 2002; 29: 193– 202 [CrossRef] [PubMed]
    [Google Scholar]
  19. Kuhlmann M, Horvay K, Strathmann A, Heinekamp T, Fischer U et al. The alpha-helical D1 domain of the tobacco bZIP transcription factor BZI-1 interacts with the ankyrin-repeat protein ANK1 and is important for BZI-1 function, both in auxin signaling and pathogen response. J Biol Chem 2003; 278: 8786– 8794 [CrossRef] [PubMed]
    [Google Scholar]
  20. Ueki S, Spektor R, Natale DM, Citovsky V. ANK, a host cytoplasmic receptor for the tobacco mosaic virus cell-to-cell movement protein, facilitates intercellular transport through plasmodesmata. PLoS Pathog 2010; 6: e1001201 [CrossRef] [PubMed]
    [Google Scholar]
  21. Emanuelsson O, Brunak S, von Heijne G, Nielsen H. Locating proteins in the cell using TargetP, SignalP and related tools. Nat Protoc 2007; 2: 953– 971 [CrossRef] [PubMed]
    [Google Scholar]
  22. Ktx V, Kim CY, Chandran AKN, Jung KH, An G et al. Molecular insights into the function of ankyrin proteins in plants. J Plant Biol 2015; 58: 271– 284 [Crossref]
    [Google Scholar]
  23. Goodin MM, Dietzgen RG, Schichnes D, Ruzin S, Jackson AO. pGD vectors: versatile tools for the expression of green and red fluorescent protein fusions in agroinfiltrated plant leaves. Plant J 2002; 31: 375– 383 [CrossRef] [PubMed]
    [Google Scholar]
  24. Kim DH, Lee JE, Xu ZY, Geem KR, Kwon Y et al. Cytosolic targeting factor AKR2A captures chloroplast outer membrane-localized client proteins at the ribosome during translation. Nat Commun 2015; 6: 6843 [CrossRef] [PubMed]
    [Google Scholar]
  25. Jarvis P. Targeting of nucleus-encoded proteins to chloroplasts in plants. New Phytol 2008; 179: 257– 285 [CrossRef] [PubMed]
    [Google Scholar]
  26. Bhattacharyya D, Chakraborty S. Chloroplast: the Trojan horse in plant-virus interaction. Mol Plant Pathol 2018; 19: 504– 518 [CrossRef] [PubMed]
    [Google Scholar]
  27. Nomura H, Komori T, Uemura S, Kanda Y, Shimotani K et al. Chloroplast-mediated activation of plant immune signalling in Arabidopsis. Nat Commun 2012; 3: 926 [CrossRef] [PubMed]
    [Google Scholar]
  28. Burch-Smith TM, Brunkard JO, Choi YG, Zambryski PC. Organelle-nucleus cross-talk regulates plant intercellular communication via plasmodesmata. Proc Natl Acad Sci USA 2011; 108: E1451 E1460 [CrossRef] [PubMed]
    [Google Scholar]
  29. Vaira AM, Maroon-Lango CJ, Hammond J. Molecular characterization of Lolium latent virus, proposed type member of a new genus in the family Flexiviridae. Arch Virol 2008; 153: 1263– 1270 [CrossRef] [PubMed]
    [Google Scholar]
  30. Johansen LK, Carrington JC. Silencing on the spot. Induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system. Plant Physiol 2001; 126: 930– 938 [CrossRef] [PubMed]
    [Google Scholar]
  31. Silhavy D, Molnár A, Lucioli A, Szittya G, Hornyik C et al. A viral protein suppresses RNA silencing and binds silencing-generated, 21- to 25-nucleotide double-stranded RNAs. Embo J 2002; 21: 3070– 3080 [CrossRef] [PubMed]
    [Google Scholar]
  32. Waadt R, Schmidt LK, Lohse M, Hashimoto K, Bock R et al. Multicolor bimolecular fluorescence complementation reveals simultaneous formation of alternative CBL/CIPK complexes in planta. Plant J 2008; 56: 505– 516 [CrossRef] [PubMed]
    [Google Scholar]
  33. Liu Y, Schiff M, Dinesh-Kumar SP. Virus-induced gene silencing in tomato. Plant J 2002; 31: 777– 786 [CrossRef] [PubMed]
    [Google Scholar]
  34. Catoni M, Lucioli A, Doblas-Ibáñez P, Accotto GP, Vaira AM. From immunity to susceptibility: virus resistance induced in tomato by a silenced transgene is lost as TGS overcomes PTGS. Plant J 2013; 75: 941– 953 [CrossRef] [PubMed]
    [Google Scholar]
  35. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2 DDCt method. Methods 2001; 25: 402– 408 [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001043
Loading
/content/journal/jgv/10.1099/jgv.0.001043
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