Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Atomic structure of potato virus X, the prototype of the Alphaflexiviridae family

Abstract

Potato virus X (PVX) is a positive-sense single-stranded RNA (ssRNA) filamentous plant virus belonging to the Alphaflexiviridae family, considered in recent years as a tool for nanotechnology applications. We present the cryo-electron microscopy structure of the PVX particle at a resolution of 2.2 Å. The well-defined density of the coat proteins and of the genomic RNA allowed a detailed analysis of protein–RNA interactions, including those mediated by solvent molecules. The particle is formed by repeated segments made of 8.8 coat proteins, forming a left-handed helical structure. The RNA runs in an internal crevice along the virion, packaged in 5-nucleotide repeats in which the first four bases are stacked in the classical way, while the fifth is rotated and nearly perpendicular. The resolution of the structure described here suggests a mechanism for the virion assembly and potentially provides a platform for the rational design of antiviral compounds and for the use of PVX in nanotechnology.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Structure of the PVX coat protein.
Fig. 2: Cryo-EM structure of PVX virus at near-atomic resolution.
Fig. 3: Interactions between CP units.
Fig. 4: Qualitative electrostatic surface of 13 units (approximately 1.5 turns) of the PVX virion.
Fig. 5: Details of PVX capsid–RNA interactions.
Fig. 6: Progression of virion assembly.

Similar content being viewed by others

Data availability

Structural data have been deposited in the PDB under accession number 6R7G and at the Electron Microscopy Data Bank (EMDB) under accession number EMD-4740. Raw data have been deposited in the Electron Microscopy Public Image Archive (EMPIAR). All other data generated or analyzed during this study are included in this published article (and its Supplementary information files) or are available from the corresponding author on reasonable request.

References

  1. Adams, M. J. et al. Virology division news: the new plant virus family Flexiviridae and assessment of molecular criteria for species demarcation. Arch. Virol. 149, 1045–1060 (2004).

    CAS  PubMed  Google Scholar 

  2. Huisman, M. J., Linthorst, H. J., Bol, J. F. & Cornelissen, J. C. The complete nucleotide sequence of potato virus X and its homologies at the amino acid level with various plus-stranded RNA viruses. J. Gen. Virol. 69, 1789–1798 (1988).

    CAS  PubMed  Google Scholar 

  3. Skryabin, K. G. et al. Conserved and variable elements in RNA genomes of potexviruses. FEBS Lett. 240, 33–40 (1988).

    CAS  PubMed  Google Scholar 

  4. Kendall, A. et al. Structure of flexible filamentous plant viruses. J. Virol. 82, 9546–9554 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Scholthof, K.-B. G. et al. Top 10 plant viruses in molecular plant pathology. Mol. Plant Pathol. 12, 938–954 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Verchot-Lubicz, J., Ye, C. M. C.-M. & Bamunusinghe, D. Molecular biology of potexviruses: recent advances. J. Gen. Virol. 88, 1643–1655 (2007).

    CAS  PubMed  Google Scholar 

  7. Röder, J., Dickmeis, C. & Commandeur, U. Small, smaller, nano: new applications for potato virus X in Nanotechnology. Front. Plant Sci. 10, 1–17 (2019).

  8. Lico, C., Schoubben, A., Baschieri, S., Blasi, P. & Santi, L. Nanoparticles in biomedicine: new insights from plant viruses. Curr. Med. Chem. 20, 3471–3487 (2013).

    CAS  PubMed  Google Scholar 

  9. Lico, C., Benvenuto, E. & Baschieri, S. The two-faced potato virus X: from plant pathogen to smart nanoparticle. Front. Plant Sci. 6, 1009 (2015).

    PubMed  PubMed Central  Google Scholar 

  10. Baratova, L. A. et al. The organization of potato virus X coat proteins in virus particles studied by tritium planigraphy and model building. Virology 188, 175–180 (1992).

    CAS  PubMed  Google Scholar 

  11. Lukashina, E. et al. Tritium planigraphy study of structural alterations in the coat protein of Potato virus X induced by binding of its triple gene block 1 protein to virions. FEBS J. 276, 7006–7015 (2009).

    CAS  PubMed  Google Scholar 

  12. Lukashina, E. et al. Analysis of the role of the coat protein N-terminal segment in potato virus X virion stability and functional activity. Mol. Plant Pathol. 13, 38–45 (2012).

    CAS  PubMed  Google Scholar 

  13. Parker, L., Kendall, A. & Stubbs, G. Surface features of potato virus X from fiber diffraction. Virology 300, 291–295 (2002).

    CAS  PubMed  Google Scholar 

  14. DiMaio, F. et al. The molecular basis for flexibility in the flexible filamentous plant viruses. Nat. Struct. Mol. Biol. 22, 642–644 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Agirrezabala, X. et al. The near-atomic cryoEM structure of a flexible filamentous plant virus shows homology of its coat protein with nucleoproteins of animal viruses. eLife 4, e11795 (2015).

    PubMed  PubMed Central  Google Scholar 

  16. Zamora, M. et al. Potyvirus virion structure shows conserved protein fold and RNA binding site in ssRNA viruses. Sci. Adv. 3, eaao2182 (2017).

    PubMed  PubMed Central  Google Scholar 

  17. Kendall, A. et al. A common structure for the potexviruses. Virology 436, 173–178 (2013).

    CAS  PubMed  Google Scholar 

  18. Valle, M. Structural homology between nucleoproteins of ssRNA viruses. Sub-Cell. Biochem. 88, 129–145 (2018).

    CAS  Google Scholar 

  19. Yang, S. et al. Crystal structure of the coat protein of the flexible filamentous papaya mosaic virus. J. Mol. Biol. 422, 263–273 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Tozzini, A. C., Ek, B., Palva, E. T. & Hopp, H. E. Potato virus X coat protein: a glycoprotein. Virology 202, 651–658 (1994).

    CAS  PubMed  Google Scholar 

  21. Baratova, L. A. et al. N-Terminal segment of potato virus X coat protein subunits is glycosylated and mediates formation of a bound water shell on the virion surface. Eur. J. Biochem. 271, 3136–3145 (2004).

    CAS  PubMed  Google Scholar 

  22. Chapman, S., Hills, G., Watts, J. & Baulcombe, D. Mutational analysis of the coat protein gene of potato virus X: effects on virion morphology and viral pathogenicity. Virology 191, 223–230 (1992).

    CAS  PubMed  Google Scholar 

  23. Betti, C. et al. Potato virus X movement in Nicotiana benthamiana: new details revealed by chimeric coat protein variants. Mol. Plant Pathol. 13, 198–203 (2012).

    CAS  PubMed  Google Scholar 

  24. Olson, W. K. Configurational statistics of polynucleotide chains. An updated virtual bond model to treat effects of base stacking. Macromolecules 13, 721–728 (1980).

    CAS  Google Scholar 

  25. Keating, K. S., Humphris, E. L. & Pyle, A. M. A new way to see RNA. Q. Rev. Biophys. 44, 433–466 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Kezar, A. et al. Structural basis for the multitasking nature of the potato virus Y coat protein. Sci. Adv. 5, 1–14 (2019).

    Google Scholar 

  27. Kaftanova, A. S., Kiselev, N. A., Novikov, V. K. & Atabekov, J. G. Structure of products of protein reassembly and reconstruction of potato virus X. Virology 65, 283–287 (1975).

    Google Scholar 

  28. Kwon, S.-J. et al. cis-Acting sequences required for coat protein binding and in vitro assembly of potato virus X. Virology 334, 83–97 (2005).

  29. Park, M.-R., Kwon, S.-J., Choi, H.-S., Hemenway, C. L. & Kim, K.-H. Mutations that alter a repeated ACCA element located at the 5′ end of the Potato virus X genome affect RNA accumulation. Virology 378, 133–141 (2008).

    CAS  PubMed  Google Scholar 

  30. Atabekov, J. G. et al. Translational activation of encapsidated potato virus X RNA by coat protein phosphorylation. Virology 286, 466–474 (2001).

    CAS  PubMed  Google Scholar 

  31. Kozlovsky, S. V. et al. Effect of the N-terminal domain of the coat protein of potato virus X on the structure of viral particles. Dokl. Biochem. Biophys. 391, 189–191 (2003).

    CAS  PubMed  Google Scholar 

  32. Le, D. H. T., Lee, K. L., Shukla, S., Commandeur, U. & Steinmetz, N. F. Potato virus X, a filamentous plant viral nanoparticle for doxorubicin delivery in cancer therapy. Nanoscale 9, 2348–2357 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Lico, C. et al. Plant-produced potato virus X chimeric particles displaying an influenza virus-derived peptide activate specific CD8+ T cells in mice. Vaccine 27, 5069–5076 (2009).

    CAS  PubMed  Google Scholar 

  34. Blandino, A. et al. In vitro and in vivo toxicity evaluation of plant virus nanocarriers. Colloids Surf. B Biointerfaces 129, 130–136 (2015).

    CAS  PubMed  Google Scholar 

  35. Lico, C. et al. A biodistribution study of two differently shaped plant virus nanoparticles reveals new peculiar traits. Colloids Surf. B Biointerfaces 148, 431–439 (2016).

    CAS  PubMed  Google Scholar 

  36. Lico, C. et al. Peptide display on Potato virus X: molecular features of the coat protein-fused peptide affecting cell-to-cell and phloem movement of chimeric virus particles. J. Gen. Virol. 87, 3103–3112 (2006).

    CAS  PubMed  Google Scholar 

  37. Vaculik, P. et al. Potato virus X displaying the E7 peptide derived from human papillomavirus type 16: a novel position for epitope presentation. Plant Cell, Tissue Organ Cult. 120, 671–680 (2015).

    CAS  Google Scholar 

  38. Cerovska, N. et al. Transient expression of Human papillomavirus type 16 L2 epitope fused to N- and C-terminus of coat protein of Potato virus X in plants. J. Biosci. 37, 125–133 (2012).

    CAS  PubMed  Google Scholar 

  39. Hoffmeisterova, H., Moravec, T., Plchova, H., Folwarczna, J. & Cerovska, N. The influence of the N- and C- terminal modifications of Potato virus X coat protein on virus properties. Biol. Plant. 56, 775–779 (2012).

    CAS  Google Scholar 

  40. Kandiah, E. et al. CM01: a facility for cryo-electron microscopy at the European Synchrotron. Acta Crystallogr. D 75, 528–535 (2019).

    CAS  Google Scholar 

  41. Li, X. et al. Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM. Nat. Methods 10, 584–590 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Zhang, K. Gctf: real-time CTF determination and correction. J. Struct. Biol. 193, 1–12 (2016).

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Zivanov, J. et al. New tools for automated high-resolution cryo-EM structure determination in RELION-3. eLife 7, e42166 (2018).

    PubMed  PubMed Central  Google Scholar 

  44. He, S. & Scheres, S. H. W. Helical reconstruction in RELION. J. Struct. Biol. 198, 163–176 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Kucukelbir, A., Sigworth, F. J. & Tagare, H. D. Quantifying the local resolution of cryo-EM density maps. Nat. Methods 11, 63–65 (2014).

    CAS  PubMed  Google Scholar 

  46. Bienert, S. et al. SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res. 46, W296–W303 (2018).

    PubMed  PubMed Central  Google Scholar 

  47. Pettersen, E. F. et al. UCSF Chimera—a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612 (2004).

    CAS  PubMed  Google Scholar 

  48. Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D 60, 2126–2132 (2004).

    PubMed  Google Scholar 

  49. Afonine, P. V. et al. New tools for the analysis and validation of cryo-EM maps and atomic models. Acta Crystallogr. D 74, 814–840 (2018).

    CAS  Google Scholar 

Download references

Acknowledgements

We acknowledge the European Synchrotron Radiation Facility for provision of beam time on CM01. We thank G. A. Leonard for support and for critically reading the manuscript.

Author information

Authors and Affiliations

Authors

Contributions

A.G. prepared grids, performed data analysis and processing, and carried out structure determination and refinement. E.K. collected images and supervised A.G. during data processing. C.L., C.B. and S.B. provided the purified virus and performed the mutagenesis experiments. S.B. and G.Z. planned the experiment. G.Z. analyzed the structure. All authors contributed to writing of the paper.

Corresponding author

Correspondence to Giuseppe Zanotti.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary Tables 1–7, Supplementary Figs 1–11

Reporting Summary

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Grinzato, A., Kandiah, E., Lico, C. et al. Atomic structure of potato virus X, the prototype of the Alphaflexiviridae family. Nat Chem Biol 16, 564–569 (2020). https://doi.org/10.1038/s41589-020-0502-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41589-020-0502-4

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing