1887

Abstract

The X protein of Borna disease virus (BDV) negatively regulates viral polymerase activity. With a BDV mini-replicon system, 30 % inhibition of polymerase activity was observed at an X to phosphoprotein (P) plasmid ratio of 1 : 6 and 100 % inhibition at a ratio of 1 : 1. It was therefore hypothesized that (i) the X : P ratio in infected cells is not significantly higher than 1 : 6 to prevent complete inhibition of polymerase activity and (ii) X is not efficiently incorporated into viral particles, allowing efficient replication early in infection. To test these assumptions, a monoclonal antibody directed against BDV X was generated. Immunofluorescence analysis revealed co-localization of X with the nucleoprotein (N) and P in the nucleus, as well as in the cytoplasm of BDV-infected cells. Quantification of viral protein levels by Western blot analysis, using purified -derived X, P and N as protein standards, revealed an X : P : N ratio in BDV-infected cells of approximately 1 : 6 : 40. However, only traces of X could be detected in purified BDV stock, suggesting that X is excluded from virus particles. These results indicate that X is a non-structural protein. The lack of X in virus particles may facilitate polymerase activity early in infection; however, the presence of X in persistently infected cells may result in partial inhibition of the polymerase and thus contribute to viral persistence.

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2005-11-01
2020-07-07
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References

  1. Bhella D., Ralph A., Murphy L. B., Yeo R. P. 2002; Significant differences in nucleocapsid morphology within the Paramyxoviridae . J Gen Virol 83:1831–1839
    [Google Scholar]
  2. Briese T., de la Torre J. C., Lewis A., Ludwig H., Lipkin W. I. 1992; Borna disease virus, a negative-strand RNA virus, transcribes in the nucleus of infected cells. Proc Natl Acad Sci U S A 89:11486–11489 [CrossRef]
    [Google Scholar]
  3. Briese T., Schneemann A., Lewis A. J., Park Y.-S., Kim S., Ludwig H., Lipkin W. I. 1994; Genomic organization of Borna disease virus. Proc Natl Acad Sci U S A 91:4362–4366 [CrossRef]
    [Google Scholar]
  4. Calain P., Roux L. 1993; The rule of six, a basic feature for efficient replication of Sendai virus defective interfering RNA. J Virol 67:4822–4830
    [Google Scholar]
  5. Cubitt B., Oldstone C., Valcarel V., de la Torre J. C. 1994; RNA splicing contributes to the generation of mature mRNAs of Borna disease virus, a non-segmented negative strand RNA virus. Virus Res 34:69–79 [CrossRef]
    [Google Scholar]
  6. Frank R. 1992; Spot-Synthesis: an easy technique for the positionally addressable, parallel chemical synthesis on a membrane support. Tetrahedron 48:9217–9232 [CrossRef]
    [Google Scholar]
  7. Geib T., Sauder C., Venturelli S., Hassler C., Staeheli P., Schwemmle M. 2003; Selective virus resistance conferred by expression of Borna disease virus nucleocapsid components. J Virol 77:4283–4290 [CrossRef]
    [Google Scholar]
  8. Hallensleben W., Schwemmle M., Hausmann J., Stitz L., Volk B., Pagenstecher A., Staeheli P. 1998; Borna disease virus-induced neurological disorder in mice: infection of neonates results in immunopathology. J Virol 72:4379–4386
    [Google Scholar]
  9. Hornig M., Briese T., Lipkin W. I. 2003; Borna disease virus. J Neurovirol 9:259–273 [CrossRef]
    [Google Scholar]
  10. Jordan I., Lipkin W. I. 2001; Borna disease virus. Rev Med Virol 11:37–57 [CrossRef]
    [Google Scholar]
  11. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. 1979; A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol 123:1548–1550
    [Google Scholar]
  12. Kobayashi T., Zhang G., Lee B. J., Baba S., Yamashita M., Kamitani W., Yanai H., Tomonaga K., Ikuta K. 2003; Modulation of Borna disease virus phosphoprotein nuclear localization by the viral protein X encoded in the overlapping open reading frame. J Virol 77:8099–8107 [CrossRef]
    [Google Scholar]
  13. Laemmli U. K., Favre M. 1973; Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol 80:575–599 [CrossRef]
    [Google Scholar]
  14. Ludwig H., Bode L., Gosztonyi G. 1988; Borna disease: a persistent disease of the central nervous system. Prog Med Virol 35:107–151
    [Google Scholar]
  15. Perez M., de la Torre J. C. 2005; Identification of the Borna disease virus (BDV) proteins required for the formation of BDV-like particles. J Gen Virol 86:1891–1895 [CrossRef]
    [Google Scholar]
  16. Perez M., Sanchez A., Cubitt B., Rosario D., de la Torre J. C. 2003; A reverse genetics system for Borna disease virus. J Gen Virol 84:3099–3104 [CrossRef]
    [Google Scholar]
  17. Planz O., Bechter K., Schwemmle M. 2002; Human borna disease virus infection. In Borna Disease Virus and Its Role in Neurobehavioral Disease pp  179–225 Edited by Carbone K. M. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  18. Pleschka S., Staeheli P., Kolodziejek J., Richt J. A., Nowotny N., Schwemmle M. 2001; Conservation of coding potential and terminal sequences in four different isolates of Borna disease virus. J Gen Virol 82:2681–2690
    [Google Scholar]
  19. Pletnikov M., Gonzalez-Dunia D., Stitz L. 2002; Experimental infection: pathogenesis of neurobehavioral disease. In Borna Disease Virus and Its Role in Neurobehavioral Disease pp  125–178 Edited by Carbone K. M. Washington, DC: American Society for Microbiology;
    [Google Scholar]
  20. Poenisch M., Unterstab G., Wolff T., Staeheli P., Schneider U. 2004; The X protein of Borna disease virus regulates viral polymerase activity through interaction with the P protein. J Gen Virol 85:1895–1898 [CrossRef]
    [Google Scholar]
  21. Schneider P. A., Schneemann A., Lipkin W. I. 1994; RNA splicing in Borna disease virus, a nonsegmented, negative-strand RNA virus. J Virol 68:5007–5012
    [Google Scholar]
  22. Schneider U., Naegele M., Staeheli P., Schwemmle M. 2003; Active Borna disease virus polymerase complex requires a distinct nucleoprotein-to-phosphoprotein ratio but no viral X protein. J Virol 77:11781–11789 [CrossRef]
    [Google Scholar]
  23. Schneider U., Blechschmidt K., Schwemmle M., Staeheli P. 2004; Overlap of interaction domains indicates a central role of the P protein in assembly and regulation of the Borna disease virus polymerase complex. J Biol Chem 279:55290–55296 [CrossRef]
    [Google Scholar]
  24. Schwemmle M., De B., Shi L., Banerjee A., Lipkin W. I. 1997; Borna disease virus P-protein is phosphorylated by protein kinase C ε and casein kinase II. J Biol Chem 272:21818–21823 [CrossRef]
    [Google Scholar]
  25. Schwemmle M., Salvatore M., Shi L., Richt J., Lee C., Lipkin W. 1998; Interactions of the Borna disease virus P, N, and X proteins and their functional implications. J Biol Chem 273:9007–9012 [CrossRef]
    [Google Scholar]
  26. Staeheli P., Sauder C., Hausmann J., Ehrensperger F., Schwemmle M. 2000; Epidemiology of Borna disease virus. J Gen Virol 81:2123–2135
    [Google Scholar]
  27. Thomas D., Newcomb W. W., Brown J. C., Wall J. S., Hainfeld J. F., Trus B. L., Steven A. C. 1985; Mass and molecular composition of vesicular stomatitis virus: a scanning transmission electron microscopy analysis. J Virol 54:598–607
    [Google Scholar]
  28. Wehner T., Ruppert A., Herden C., Frese K., Becht H., Richt J. A. 1997; Detection of a novel Borna disease virus encoded 10 kilodalton protein in infected cells and tissues. J Gen Virol 78:2459–2466
    [Google Scholar]
  29. Wolff T., Pfleger R., Wehner T., Reinhardt J., Richt J. A. 2000; A short leucine-rich sequence in the Borna disease virus p10 protein mediates association with the viral phospho- and nucleoproteins. J Gen Virol 81:939–947
    [Google Scholar]
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