1887

Abstract

The human respiratory syncytial virus (HRSV) P protein is phosphorylated, with different turnover rates, at several serine (S) and threonine (T) residues. The role of phosphothreonines in viral RNA synthesis was studied by using P protein substitution variants and the HRSV-based minigenome pM/SH. By using liquid chromatography coupled to ion-trap mass spectrometry, it was found that P protein T108 was phosphorylated by addition of a high-turnover phosphate group. This phosphorylation occurs in P protein expressed transiently and during HRSV infection. The results suggest that phosphorylation at P protein T108 affects M2-1 transcriptional activities, because this modification prevents interaction between the P and M2-1 proteins. Therefore, P protein phosphorylation–dephosphorylation at T108 could distinguish the role of the P protein in viral transcription and replication.

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2006-12-01
2019-11-13
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References

  1. Asenjo, A., Rodríguez, L. & Villanueva, N. ( 2005; ). Determination of phosphorylated residues from human respiratory syncytial virus P protein that are dynamically dephosphorylated by cellular phosphatases: a possible role for serine 54. J Gen Virol 86, 1109–1120.[CrossRef]
    [Google Scholar]
  2. Bialojan, C. & Takai, A. ( 1988; ). Inhibitory effect of a marine-sponge toxin, okadaic acid, on protein phosphatases. Specificity and kinetics. Biochem J 256, 283–290.
    [Google Scholar]
  3. Bitko, V., Musiyenko, A., Shulyayeva, O. & Barik, S. ( 2005; ). Inhibition of respiratory viruses by nasally administered siRNA. Nat Med 11, 50–55.[CrossRef]
    [Google Scholar]
  4. Cartee, T. L. & Wertz, G. W. ( 2001; ). Respiratory syncytial virus M2-1 protein requires phosphorylation for efficient function and binds viral RNA during infection. J Virol 75, 12188–12197.[CrossRef]
    [Google Scholar]
  5. Cartee, T. L., Megaw, A. G., Oomens, A. G. P. & Wertz, G. W. ( 2003; ). Identification of a single amino acid change in the human respiratory syncytial virus L protein that affects transcriptional termination. J Virol 77, 7352–7360.[CrossRef]
    [Google Scholar]
  6. Collins, P. L., Hill, M. G., Camargo, E., Grosfeld, H., Chanock, R. M. & Murphy, B. R. ( 1995; ). Production of infectious human respiratory syncytial virus from cloned cDNA confirms an essential role for the transcription elongation factor from the 5′ proximal open reading frame of the M2 mRNA in gene expression and provides a capability for vaccine development. Proc Natl Acad Sci USA 92, 11563–11567.[CrossRef]
    [Google Scholar]
  7. Collins, P. L., Whitehead, S. S., Bukreyev, A., Fearns, R., Teng, M. N., Juhasz, K., Chanock, R. M. & Murphy, B. R. ( 1999; ). Rational design of live-attenuated recombinant vaccine virus for human respiratory syncytial virus by reverse genetics. Adv Virus Res 54, 423–451.
    [Google Scholar]
  8. Collins, P. L., Chanock, R. M. & Murphy, B. R. ( 2001; ). Respiratory syncytial virus. In Fields Virology, 4th edn, pp. 1443–1485. Edited by D. M. Knipe & P. M. Howley. Philadelphia, PA: Lippincott Williams & Wilkins.
  9. Cuesta, I., Geng, X., Asenjo, A. & Villanueva, N. ( 2000; ). Structural phosphoprotein M2-1 of the human respiratory syncytial virus is an RNA binding protein. J Virol 74, 9858–9867.[CrossRef]
    [Google Scholar]
  10. Gubbay, O., Curran, J. & Kolakofsky, D. ( 2001; ). Sendai virus genome synthesis and assembly are coupled: a possible mechanism to promote viral RNA polymerase processivity. J Gen Virol 82, 2895–2903.
    [Google Scholar]
  11. Hall, C. B. ( 2001; ). Respiratory syncytial virus and parainfluenza virus. N Engl J Med 344, 1917–1928.[CrossRef]
    [Google Scholar]
  12. Hardy, R. W. & Wertz, G. W. ( 1998; ). The product of the respiratory syncytial virus M2 gene ORF1 enhances readthrough of intergenic junctions during viral transcription. J Virol 72, 520–526.
    [Google Scholar]
  13. Hardy, R. W. & Wertz, G. W. ( 2000; ). The Cys3-His1 motif of the respiratory syncytial virus M2-1 protein is essential for protein function. J Virol 74, 5880–5885.[CrossRef]
    [Google Scholar]
  14. Lamb, R. A. & Kolakofsky, D. ( 2001; ). Paramyxoviridae: the viruses and their replication. In Fields Virology, 4th edn, pp. 1305–1340. Edited by D. M. Knipe & P. M. Howley. Philadelphia, PA: Lippincott Williams & Wilkins.
  15. Lu, B., Ma, C.-H., Brazas, R. & Jin, H. ( 2002; ). The major phosphorylation sites of the respiratory syncytial virus phosphoprotein are dispensable for virus replication in vitro. J Virol 76, 10776–10784.[CrossRef]
    [Google Scholar]
  16. Mason, S. W., Aberg, E., Lawetz, C., DeLong, R., Whitehead, P. & Liuzzi, M. ( 2003; ). Interaction between human respiratory syncytial virus (RSV) M2-1 and P proteins is required for reconstitution of M2-1-dependent RSV minigenome activity. J Virol 77, 10670–10676.[CrossRef]
    [Google Scholar]
  17. Mazumder, B., Adhikary, G. & Barik, S. ( 1994; ). Bacterial expression of human respiratory syncytial viral phosphoprotein P and identification of Ser237 as the site of phosphorylation by cellular casein kinase II. Virology 205, 93–103.[CrossRef]
    [Google Scholar]
  18. Meissner, H. C., Welliver, R. C., Chartrand, S. A., Law, B. J., Weisman, L. E., Dorkin, H. L. & Rodriguez, W. J. ( 1999; ). Immunoprophylaxis with palivizumab, a humanized respiratory syncytial virus monoclonal antibody, for prevention of respiratory syncytial virus infection in high risk infants: a consensus opinion. Pediatr Infect Dis J 18, 223–231.[CrossRef]
    [Google Scholar]
  19. Navarro, J., López-Otín, C. & Villanueva, N. ( 1991; ). Location of phosphorylated residues in human respiratory syncytial virus phosphoprotein. J Gen Virol 72, 1455–1459.[CrossRef]
    [Google Scholar]
  20. Pastey, M. K., Gower, T. L., Spearman, P. W., Crowe, J. E., Jr & Graham, B. S. ( 2000; ). A RhoA-derived peptide inhibits syncytium formation induced by respiratory syncytial virus and parainfluenza virus type 3. Nat Med 6, 35–40.[CrossRef]
    [Google Scholar]
  21. Pearson, R. B. & Kemp, B. E. ( 1991; ). Protein kinase phosphorylation site sequences and consensus specificity motifs: tabulations. Methods Enzymol 200, 62–81.
    [Google Scholar]
  22. Sánchez-Seco, M. P., Navarro, J., Martínez, R. & Villanueva, N. ( 1995; ). C-Terminal phosphorylation of human respiratory syncytial virus P protein occurs mainly at serine residue 232. J Gen Virol 76, 425–430.[CrossRef]
    [Google Scholar]
  23. Villanueva, N., Hardy, R., Asenjo, A., Yu, Q. & Wertz, G. ( 2000; ). The bulk of the phosphorylation of human respiratory syncytial virus phosphoprotein is not essential but modulates viral RNA transcription and replication. J Gen Virol 81, 129–133.
    [Google Scholar]
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