1887

Abstract

Efficient replication of varicella-zoster virus (VZV) in cell culture requires expression of protein encoded by VZV open reading frame 63 (ORF63p). Two-dimensional gel analysis demonstrates that ORF63p is extensively modified. Mass spectroscopy analysis of ORF63p isolated from transiently transfected HEK 293 and stably transfected MeWo cells identified 10 phosphorylated residues. In VZV-infected MeWo cells, only six phosphorylated residues were detected. This report identifies phosphorylation of two previously uncharacterized residues (Ser5 and Ser31) in ORF63p extracted from cells infected with VZV or transfected with an ORF63p expression plasmid. Computational analysis of ORF63p for known kinase substrates did not identify Ser5 or Ser31 as candidate phosphorylation sites, suggesting that either atypical recognition sequences or novel cellular kinases are involved in ORF63p post-translational modification.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.019067-0
2010-05-01
2019-11-12
Loading full text...

Full text loading...

/deliver/fulltext/jgv/91/5/1133.html?itemId=/content/journal/jgv/10.1099/vir.0.019067-0&mimeType=html&fmt=ahah

References

  1. Ambagala, A. P. & Cohen, J. I. ( 2007; ). Varicella-zoster virus IE63, a major viral latency protein, is required to inhibit the alpha interferon-induced antiviral response. J Virol 81, 7844–7851.[CrossRef]
    [Google Scholar]
  2. Ambagala, A. P., Bosma, T., Ali, M. A., Poustovoitov, M., Chen, J. J., Gershon, M. D., Adams, P. D. & Cohen, J. I. ( 2009; ). Varicella-zoster virus immediate-early 63 protein interacts with human antisilencing function 1 protein and alters its ability to bind histones h3.1 and h3.3. J Virol 83, 200–209.[CrossRef]
    [Google Scholar]
  3. Baiker, A., Bagowski, C., Ito, H., Sommer, M., Zerboni, L., Fabel, K., Hay, J., Ruyechan, W. T. & Arvin, A. M. ( 2004; ). The immediate-early 63 protein of varicella-zoster virus: analysis of functional domains required for replication in vitro and for T-cell and skin tropism in the SCIDhu model in vivo. J Virol 78, 1181–1194.[CrossRef]
    [Google Scholar]
  4. Bontems, S., Di Valentin, E., Baudoux, L., Rentier, B., Sadzot-Delvaux, C. & Piette, J. ( 2002; ). Phosphorylation of varicella-zoster virus IE63 protein by casein kinases influences its cellular localization and gene regulation activity. J Biol Chem 277, 21050–21060.[CrossRef]
    [Google Scholar]
  5. Chen, J. J., Gershon, A. A., Li, Z. S., Lungu, O. & Gershon, M. D. ( 2003; ). Latent and lytic infection of isolated guinea pig enteric ganglia by varicella zoster virus. J Med Virol 70, S71–S78.[CrossRef]
    [Google Scholar]
  6. Cohen, J. I., Cox, E., Pesnicak, L., Srinivas, S. & Krogmann, T. ( 2004; ). The varicella-zoster virus open reading frame 63 latency-associated protein is critical for establishment of latency. J Virol 78, 11833–11840.[CrossRef]
    [Google Scholar]
  7. Cohen, J. I., Krogmann, T., Bontems, S., Sadzot-Delvaux, C. & Pesnicak, L. ( 2005; ). Regions of the varicella-zoster virus open reading frame 63 latency-associated protein important for replication in vitro are also critical for efficient establishment of latency. J Virol 79, 5069–5077.[CrossRef]
    [Google Scholar]
  8. Cohrs, R. J. & Gilden, D. H. ( 2007; ). Prevalence and abundance of latently transcribed varicella-zoster virus genes in human ganglia. J Virol 81, 2950–2956.[CrossRef]
    [Google Scholar]
  9. Cohrs, R. J., Barbour, M. & Gilden, D. H. ( 1996; ). Varicella-zoster virus (VZV) transcription during latency in human ganglia: detection of transcripts mapping to genes 21, 29, 62, and 63 in a cDNA library enriched for VZV RNA. J Virol 70, 2789–2796.
    [Google Scholar]
  10. Cohrs, R. J., Randall, J., Smith, J., Gilden, D. H., Dabrowski, C., van Der, K. H. & Tal-Singer, R. ( 2000; ). Analysis of individual human trigeminal ganglia for latent herpes simplex virus type 1 and varicella-zoster virus nucleic acids using real-time PCR. J Virol 74, 11464–11471.[CrossRef]
    [Google Scholar]
  11. Cohrs, R. J., Wischer, J., Essman, C. & Gilden, D. H. ( 2002; ). Characterization of varicella-zoster virus gene 21 and 29 proteins in infected cells. J Virol 76, 7228–7238.[CrossRef]
    [Google Scholar]
  12. Debrus, S., Sadzot-Delvaux, C., Nikkels, A. F., Piette, J. & Rentier, B. ( 1995; ). Varicella-zoster virus gene 63 encodes an immediate-early protein that is abundantly expressed during latency. J Virol 69, 3240–3245.
    [Google Scholar]
  13. Di Valentin, E., Bontems, S., Habran, L., Jolois, O., Markine-Goriaynoff, N., Vanderplasschen, A., Sadzot-Delvaux, C. & Piette, J. ( 2005; ). Varicella-zoster virus IE63 protein represses the basal transcription machinery by disorganizing the pre-initiation complex. Biol Chem 386, 255–267.
    [Google Scholar]
  14. Gary, L., Gilden, D. H. & Cohrs, R. J. ( 2006; ). Epigenetic regulation of varicella-zoster virus open reading frames 62 and 63 in latently infected human trigeminal ganglia. J Virol 80, 4921–4926.[CrossRef]
    [Google Scholar]
  15. Gilden, D. H., Cohrs, R. J. & Mahalingam, R. ( 2003; ). Clinical and molecular pathogenesis of varicella virus infection. Viral Immunol 16, 243–258.[CrossRef]
    [Google Scholar]
  16. Grinfeld, E. & Kennedy, P. G. ( 2004; ). Translation of varicella-zoster virus genes during human ganglionic latency. Virus Genes 29, 317–319.[CrossRef]
    [Google Scholar]
  17. Grose, C. & Brunel, P. A. ( 1978; ). Varicella-zoster virus: isolation and propagation in human melanoma cells at 36 and 32 degrees C. Infect Immun 19, 199–203.
    [Google Scholar]
  18. Habran, L., Bontems, S., Di Valentin, E., Sadzot-Delvaux, C. & Piette, J. ( 2005; ). Varicella-zoster virus IE63 protein phosphorylation by roscovitine-sensitive cyclin-dependent kinases modulates its cellular localization and activity. J Biol Chem 280, 29135–29143.[CrossRef]
    [Google Scholar]
  19. Heineman, T. C. & Cohen, J. I. ( 1995; ). The varicella-zoster virus (VZV) open reading frame 47 (ORF47) protein kinase is dispensable for viral replication and is not required for phosphorylation of ORF63 protein, the VZV homolog of herpes simplex virus ICP22. J Virol 69, 7367–7370.
    [Google Scholar]
  20. Heineman, T. C., Seidel, K. & Cohen, J. I. ( 1996; ). The varicella-zoster virus ORF66 protein induces kinase activity and is dispensable for viral replication. J Virol 70, 7312–7317.
    [Google Scholar]
  21. Jackers, P., Defechereux, P., Baudoux, L., Lambert, C., Massaer, M., Merville-Louis, M. P., Rentier, B. & Piette, J. ( 1992; ). Characterization of regulatory functions of the varicella-zoster virus gene 63-encoded protein. J Virol 66, 3899–3903.
    [Google Scholar]
  22. Kennedy, P. G., Grinfeld, E. & Bell, J. E. ( 2000; ). Varicella-zoster virus gene expression in latently infected and explanted human ganglia. J Virol 74, 11893–11898.[CrossRef]
    [Google Scholar]
  23. Kennedy, P. G., Grinfeld, E., Bontems, S. & Sadzot-Delvaux, C. ( 2001; ). Varicella-zoster virus gene expression in latently infected rat dorsal root ganglia. Virology 289, 218–223.[CrossRef]
    [Google Scholar]
  24. Kenyon, T. K., Lynch, J., Hay, J., Ruyechan, W. & Grose, C. ( 2001; ). Varicella-zoster virus ORF47 protein serine kinase: characterization of a cloned, biologically active phosphotransferase and two viral substrates, ORF62 and ORF63. J Virol 75, 8854–8858.[CrossRef]
    [Google Scholar]
  25. Kinchington, P. R., Bookey, D. & Turse, S. E. ( 1995; ). The transcriptional regulatory proteins encoded by varicella-zoster virus open reading frames (ORFs) 4 and 63, but not ORF 61, are associated with purified virus particles. J Virol 69, 4274–4282.
    [Google Scholar]
  26. Kost, R. G., Kupinsky, H. & Straus, S. E. ( 1995; ). Varicella-zoster virus gene 63: transcript mapping and regulatory activity. Virology 209, 218–224.[CrossRef]
    [Google Scholar]
  27. Lungu, O., Panagiotidis, C. A., Annunziato, P. W., Gershon, A. A. & Silverstein, S. J. ( 1998; ). Aberrant intracellular localization of varicella-zoster virus regulatory proteins during latency. Proc Natl Acad Sci U S A 95, 7080–7085.[CrossRef]
    [Google Scholar]
  28. Lynch, J. M., Kenyon, T. K., Grose, C., Hay, J. & Ruyechan, W. T. ( 2002; ). Physical and functional interaction between the varicella zoster virus IE63 and IE62 proteins. Virology 302, 71–82.[CrossRef]
    [Google Scholar]
  29. Mahalingam, R., Wellish, M., Cohrs, R., Debrus, S., Piette, J., Rentier, B. & Gilden, D. H. ( 1996; ). Expression of protein encoded by varicella-zoster virus open reading frame 63 in latently infected human ganglionic neurons. Proc Natl Acad Sci U S A 93, 2122–2124.[CrossRef]
    [Google Scholar]
  30. Messaoudi, I., Barron, A., Wellish, M., Engelmann, F., Legasse, A., Planer, S., Gilden, D., Nikolich-Zugich, J. & Mahalingam, R. ( 2009; ). Simian varicella virus infection of rhesus macaques recapitulates essential features of varicella zoster virus infection in humans. PLoS Pathog 5, e1000657 [CrossRef]
    [Google Scholar]
  31. Mueller, N. H., Graf, L. L., Orlicky, D., Gilden, D. & Cohrs, R. J. ( 2009; ). Phosphorylation of the nuclear form of varicella zoster virus immediate-early protein 63 by casein kinase II at serine 186. J Virol 83, 12094–12100.[CrossRef]
    [Google Scholar]
  32. Stevenson, D., Xue, M., Hay, J. & Ruyechan, W. T. ( 1996; ). Phosphorylation and nuclear localization of the varicella-zoster virus gene 63 protein. J Virol 70, 658–662.
    [Google Scholar]
  33. Stolarov, J., Chang, K., Reiner, A., Rodgers, L., Hannon, G. J., Wigler, M. H. & Mittal, V. ( 2001; ). Design of a retroviral-mediated ecdysone-inducible system and its application to the expression profiling of the PTEN tumor suppressor. Proc Natl Acad Sci U S A 98, 13043–13048.[CrossRef]
    [Google Scholar]
  34. Walters, M. S., Kyratsous, C. A., Wan, S. & Silverstein, S. ( 2008; ). Nuclear import of the varicella-zoster virus latency-associated protein ORF63 in primary neurons requires expression of the lytic protein ORF61 and occurs in a proteasome-dependent manner. J Virol 82, 8673–8686.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.019067-0
Loading
/content/journal/jgv/10.1099/vir.0.019067-0
Loading

Data & Media loading...

Supplements

vol. , part 5, pp. 1133 - 1137

Mass spectral analysis of ORF63p peptide

ORF63p expression analysis in ecdysone inducible cell line [Single PDF file](238 KB)



PDF

Most Cited This Month

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