1887

Abstract

The NS3 protein of rice hoja blanca virus represents a viral suppressor of RNA interference (RNAi) that sequesters small interfering (si)RNAs . To determine whether this siRNA binding property is the critical determinant for the suppressor activity of NS3, NS3 was altered by alanine point mutations and the resulting mutant proteins were tested for both siRNA binding ability and RNAi suppressor activity in plants. Alanine substitutions of lysine residues at positions 173–175 resulted in mutant proteins that lost both their affinity for siRNAs and their RNAi suppressor activity . This indicates that siRNA binding of NS3 is indeed essential for the suppressor function of NS3 and that residues at positions 173–175 are involved in the siRNA binding and suppressor activities.

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2009-07-01
2020-01-29
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References

  1. Brodersen, P. & Voinnet, O. ( 2006; ). The diversity of RNA silencing pathways in plants. Trends Genet 22, 268–280.[CrossRef]
    [Google Scholar]
  2. Bucher, E., Sijen, T., de Haan, P., Goldbach, R. & Prins, M. ( 2003; ). Negative-strand tospoviruses and tenuiviruses carry a gene for a suppressor of gene silencing at analogous genomic positions. J Virol 77, 1329–1336.[CrossRef]
    [Google Scholar]
  3. Bucher, E., Hemmes, H., de Haan, P., Goldbach, R. & Prins, M. ( 2004; ). The influenza A virus NS1 protein binds small interfering RNAs and suppresses RNA silencing in plants. J Gen Virol 85, 983–991.[CrossRef]
    [Google Scholar]
  4. Chu, M., Desvoyes, B., Turina, M., Noad, R. & Scholthof, H. B. ( 2000; ). Genetic dissection of tomato bushy stunt virus p19-protein-mediated host-dependent symptom induction and systemic invasion. Virology 266, 79–87.[CrossRef]
    [Google Scholar]
  5. Ding, S. W. & Voinnet, O. ( 2007; ). Antiviral immunity directed by small RNAs. Cell 130, 413–426.[CrossRef]
    [Google Scholar]
  6. Falk, B. W. & Tsai, J. H. ( 1998; ). Biology and molecular biology of viruses in the genus Tenuivirus. Annu Rev Phytopathol 36, 139–163.[CrossRef]
    [Google Scholar]
  7. Haasnoot, J., de Vries, W., Geutjes, E. J., Prins, M., de Haan, P. & Berkhout, B. ( 2007; ). The Ebola virus VP35 protein is a suppressor of RNA silencing. PLoS Pathog 3, e86 [CrossRef]
    [Google Scholar]
  8. Hartman, A. L., Towner, J. S. & Nichol, S. T. ( 2004; ). A C-terminal basic amino acid motif of Zaire ebolavirus VP35 is essential for type I interferon antagonism and displays high identity with the RNA-binding domain of another interferon antagonist, the NS1 protein of influenza A virus. Virology 328, 177–184.[CrossRef]
    [Google Scholar]
  9. Hemmes, H., Lakatos, L., Goldbach, R., Burgyán, J. & Prins, M. ( 2007; ). The NS3 protein of Rice hoja blanca tenuivirus suppresses RNA silencing in plant and insect hosts by efficiently binding both siRNAs and miRNAs. RNA 13, 1079–1089.[CrossRef]
    [Google Scholar]
  10. Karimi, M., Inze, D. & Depicker, A. ( 2002; ). GATEWAY vectors for Agrobacterium-mediated plant transformation. Trends Plant Sci 7, 193–195.[CrossRef]
    [Google Scholar]
  11. Lakatos, L., Csorba, T., Pantaleo, V., Chapman, E. J., Carrington, J. C., Liu, Y. P., Dolja, V. V., Calvino, L. F., López-Moya, J. J. & Burgyán, J. ( 2006; ). Small RNA binding is a common strategy to suppress RNA silencing by several viral suppressors. EMBO J 25, 2768–2780.[CrossRef]
    [Google Scholar]
  12. Mérai, Z., Kerényi, Z., Kertész, S., Magna, M., Lakatos, L. & Silhavy, D. ( 2006; ). Double-stranded RNA binding may be a general plant RNA viral strategy to suppress RNA silencing. J Virol 80, 5747–5756.[CrossRef]
    [Google Scholar]
  13. Ramirez, B. C., Macaya, G., Calvert, L. A. & Haenni, A. L. ( 1992; ). Rice hoja blanca virus genome characterization and expression in vitro. J Gen Virol 73, 1457–1464.[CrossRef]
    [Google Scholar]
  14. Schnettler, E., Hemmes, H., Goldbach, R. & Prins, M. ( 2008; ). The NS3 protein of rice hoja blanca virus suppresses RNA silencing in mammalian cells. J Gen Virol 89, 336–340.[CrossRef]
    [Google Scholar]
  15. Tomari, Y. & Zamore, P. D. ( 2005; ). Perspective: machines for RNAi. Genes Dev 19, 517–529.[CrossRef]
    [Google Scholar]
  16. van Rij, R. P., Saleh, M. C., Berry, B., Foo, C., Houk, A., Antoniewski, C. & Andino, R. ( 2006; ). The RNA silencing endonuclease Argonaute 2 mediates specific antiviral immunity in Drosophila melanogaster. Genes Dev 20, 2985–2995.[CrossRef]
    [Google Scholar]
  17. Vargason, J. M., Szittya, G., Burgyán, J. & Hall, T. M. ( 2003; ). Size selective recognition of siRNA by an RNA silencing suppressor. Cell 115, 799–811.[CrossRef]
    [Google Scholar]
  18. Vaucheret, H. ( 2006; ). Post-transcriptional small RNA pathways in plants: mechanisms and regulations. Genes Dev 20, 759–771.[CrossRef]
    [Google Scholar]
  19. Voinnet, O. ( 2005; ). Induction and suppression of RNA silencing: insights from viral infections. Nat Rev Genet 6, 206–220.[CrossRef]
    [Google Scholar]
  20. Wang, W., Riedel, K., Lynch, P., Chien, C. Y., Montelione, G. T. & Krug, R. M. ( 1999; ). RNA binding by the novel helical domain of the influenza virus NS1 protein requires its dimer structure and a small number of specific basic amino acids. RNA 5, 195–205.[CrossRef]
    [Google Scholar]
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