1887

Abstract

The icosahedral capsid of cowpea mosaic virus is formed by 60 copies of the large (L) and small (S) coat protein subunits. The 24-amino-acid C-terminal peptide of the S coat protein can undergo proteolytic cleavage without affecting particle stability or infectivity. Mutagenic studies have shown that this sequence is involved in particle assembly, virus movement, RNA encapsidation and suppression of gene silencing. However, it is unclear how these processes are related, and which part(s) of the sequence are involved in each process. Here, we have analysed the effect of mutations in the C-terminal region of the S protein on the assembly of empty virus-like particles and on the systemic movement of infectious virus. The results confirmed the importance of positively charged amino acids adjacent to the cleavage site for particle assembly and revealed that the C-terminal 11 amino acids are important for efficient systemic movement of the virus.

  • This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001285
2019-06-06
2024-05-17
Loading full text...

Full text loading...

/deliver/fulltext/jgv/100/7/1165.html?itemId=/content/journal/jgv/10.1099/jgv.0.001285&mimeType=html&fmt=ahah

References

  1. Franssen H, Goldbach R, Broekhuijsen M, Moerman M, van Kammen A. Expression of middle-component RNA of cowpea mosaic virus: in vitro generation of a precursor to both capsid proteins by a Bottom-Component RNA-Encoded protease from infected cells. J Virol 1982; 41:8–17
    [Google Scholar]
  2. Saunders K, Sainsbury F, Lomonossoff GP. Efficient generation of cowpea mosaic virus empty virus-like particles by the proteolytic processing of precursors in insect cells and plants. Virology 2009; 393:329–337 [View Article]
    [Google Scholar]
  3. Lin T, Chen Z, Usha R, Stauffacher CV, Dai JB et al. The refined crystal structure of cowpea mosaic virus at 2.8 A resolution. Virology 1999; 265:20–34 [View Article]
    [Google Scholar]
  4. Semancik JS. Studies on electrophoretic heterogeneity in isometric plant viruses. Virology 1966; 30:698–704 [View Article]
    [Google Scholar]
  5. Lomonossoff GP, Johnson JE. The synthesis and structure of comovirus capsids. Prog Biophys Mol Biol 1991; 55:107–137 [View Article]
    [Google Scholar]
  6. Taylor KM, Spall VE, Butler PJ, Lomonossoff GP. The cleavable carboxyl-terminus of the small coat protein of cowpea mosaic virus is involved in RNA encapsidation. Virology 1999; 255:129–137 [View Article]
    [Google Scholar]
  7. Huynh NT, Hesketh EL, Saxena P, Meshcheriakova Y, Ku Y-C et al. Crystal structure and proteomics analysis of empty virus-like particles of cowpea mosaic virus. Structure 2016; 24:567–575 [View Article]
    [Google Scholar]
  8. Hesketh EL, Meshcheriakova Y, Dent KC, Saxena P, Thompson RF et al. Mechanisms of assembly and genome packaging in an RNA virus revealed by high-resolution cryo-EM. Nat Commun 2015; 6:10113 [View Article]
    [Google Scholar]
  9. Geelen JL, Rezelman G, van Kammen A. The infectivity of the two electrophoretic forms of cowpea mosaic virus. Virology 1973; 51:279–286 [View Article]
    [Google Scholar]
  10. Theuri JM, Sehgal P. In plantabehaviour of bean pod mottle and cowpea mosaic comoviruses: infectivity and structural alterations. Physiol Mol Plant Pathol 1996; 48:193–208 [View Article]
    [Google Scholar]
  11. Cañizares MC, Taylor KM, Lomonossoff GP. Surface-exposed C-terminal amino acids of the small coat protein of cowpea mosaic virus are required for suppression of silencing. J Gen Virol 2004; 85:3431–3435 [View Article]
    [Google Scholar]
  12. Sainsbury F, Saunders K, Aljabali AAA, Evans DJ, Lomonossoff GP. Peptide-controlled access to the interior surface of empty virus nanoparticles. Chembiochem 2011; 12:2435–2440 [View Article]
    [Google Scholar]
  13. Meshcheriakova Y, Durrant A, Hesketh EL, Ranson NA, Lomonossoff GP. Combining high-resolution cryo-electron microscopy and mutagenesis to develop cowpea mosaic virus for bionanotechnology. Biochem Soc Trans 2017; 45:1263–1269 [View Article]
    [Google Scholar]
  14. Verver J, Wellink J, Van Lent J, Gopinath K, van Kammen A. Studies on the movement of cowpea mosaic virus using the jellyfish green fluorescent protein. Virology 1998; 242:22–27 [View Article]
    [Google Scholar]
  15. Gopinath K, Wellink J, Porta C, Taylor KM, Lomonossoff GP et al. Engineering cowpea mosaic virus RNA-2 into a vector to express heterologous proteins in plants. Virology 2000; 267:159–173 [View Article]
    [Google Scholar]
  16. Sainsbury F, Saxena P, Aljabali AAA, Saunders K, Evans DJ et al. Genetic engineering and characterization of cowpea mosaic virus empty virus-like particles. Meth. Mol Biol 2014; 1108:139–153
    [Google Scholar]
  17. Liu L, Lomonossoff GP. Agroinfection as a rapid method for propagating cowpea mosaic virus-based constructs. J Virol Meth 2002; 105:343–348 [View Article]
    [Google Scholar]
  18. Porta C, Spall VE, Findlay KC, Gergerich RC, Farrance CE et al. Cowpea mosaic virus-based chimaeras. Effects of inserted peptides on the phenotype, host-range and transmissibility of the modified viruses. Virology 2003; 310:50–63
    [Google Scholar]
  19. Montague NP, Thuenemann EC, Saxena P, Saunders K, Lenzi P et al. Recent advances of cowpea mosaic virus-based particle technology. Hum Vaccin 2011; 7:383–390 [View Article]
    [Google Scholar]
  20. Wellink J, van Kammen A. Cell-to-cell transport of cowpea mosaic virus requires both the 58K/48K proteins and the capsid proteins. J Gen Virol 1989; 70:2279–2286 [View Article]
    [Google Scholar]
  21. van Lent J, Wellink J, Goldbach R. Evidence for the involvement of the 58K and 48K proteins in the intercellular movement of cowpea mosaic virus. J Gen Virol 1990; 71:219–223 [View Article]
    [Google Scholar]
  22. Lekkerkerker A, Wellink J, Yuan P, van Lent J, Goldbach R et al. Distinct functional domains in the cowpea mosaic virus movement protein. J Virol 1996; 70:5658–5661
    [Google Scholar]
  23. Carvalho CM, Wellink J, Ribeiro SG, Goldbach RW, Van Lent JWM. The C-terminal region of the movement protein of cowpea mosaic virus is involved in binding to the large but not to the small coat protein. J Gen Virol 2003; 84:2271–2277 [View Article]
    [Google Scholar]
  24. Bertens P, Heijne W, van der Wel N, Wellink J, van Kammen A. Studies on the C-terminus of the cowpea mosaic virus movement protein. Arch Virol 2003; 148:265–279 [View Article]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001285
Loading
/content/journal/jgv/10.1099/jgv.0.001285
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
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