1887

Abstract

The UL47 gene product, VP8, is one of the major tegument proteins of bovine herpesvirus 1 (BoHV-1) and is subject to phosphorylation. Analysis of protein bands co-immunoprecipitated with VP8 from BoHV-1-infected cells by mass spectroscopy suggested that VP8 interacts with two protein kinases: cellular CK2 and viral US3. CK2 is a highly conserved cellular protein, expressed ubiquitously and known to phosphorylate numerous proteins. The US3 gene product is one of the viral kinases produced by BoHV-1 during infection. Interactions of CK2 and US3 with VP8 were confirmed outside the context of infection when FLAG–VP8 was expressed alone or co-expressed with US3–haemagglutinin tag in Cos-7 cells. Furthermore, VP8 and US3 were found to co-localize in the nucleus during viral infection. To explore the significance of these interactions, an kinase assay was performed, which demonstrated that VP8 is heavily phosphorylated by CK2. In the presence of the highly specific CK2 kinase inhibitor 2-dimethylamino-4,5,6,7-tetrabromo-1-benzimidazole (DMAT), phosphorylation of VP8 was significantly reduced. Phosphorylation of VP8 was also inhibited by the presence of kenpaullone, a less specific CK2 inhibitor, but not by protein kinase CK1 or protein kinase C inhibitors. When VP8 and US3 were both included in the kinase assay in the presence of DMAT, phosphorylation of VP8 was again observed. Autophosphorylation of US3 was also detected and was not inhibited by DMAT. Based on these results, it is proposed that VP8 interacts with cellular CK2 and viral US3 in BoHV-1-infected cells, and is in turn subject to kinase activities associated with both of these proteins.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.013532-0
2009-12-01
2024-11-10
Loading full text...

Full text loading...

/deliver/fulltext/jgv/90/12/2829.html?itemId=/content/journal/jgv/10.1099/vir.0.013532-0&mimeType=html&fmt=ahah

References

  1. Alvisi G., Rawlinson S. M., Ghildyal R., Ripalti A., Jans D. A. 2008; Regulated nucleocytoplasmic trafficking of viral gene products: a therapeutic target?. Biochim Biophys Acta 1784213–227 [CrossRef]
    [Google Scholar]
  2. Birchall A. M., Bishop J., Bradshaw D., Cline A., Coffey J., Elliott L. H., Gibson V. M., Greenham A., Hallam T. J. other authors 1994; Ro 32-0432, a selective and orally active inhibitor of protein kinase C prevents T-cell activation. J Pharmacol Exp Ther 268:922–929
    [Google Scholar]
  3. Carpenter D. E., Misra V. 1991; The most abundant protein in bovine herpes 1 virions is a homologue of herpes simplex virus type 1 UL47. J Gen Virol 72:3077–3084 [CrossRef]
    [Google Scholar]
  4. Cook I. D., Shanahan F., Farrell P. J. 1994; Epstein–Barr virus SM protein. Virology 205:217–227 [CrossRef]
    [Google Scholar]
  5. Daikoku T., Yamashita Y., Tsurumi T., Maeno K., Nishiyama Y. 1993; Purification and biochemical characterization of the protein kinase encoded by the US3 gene of herpes simplex virus type 2. Virology 197:685–694 [CrossRef]
    [Google Scholar]
  6. Donnelly M., Elliott G. 2001a; Fluorescent tagging of herpes simplex virus tegument protein VP13/14 in virus infection. J Virol 75:2575–2583 [CrossRef]
    [Google Scholar]
  7. Donnelly M., Elliott G. 2001b; Nuclear localization and shuttling of herpes simplex virus tegument protein VP13/14. J Virol 75:2566–2574 [CrossRef]
    [Google Scholar]
  8. Donnelly M., Verhagen J., Elliott G. 2007; RNA binding by the herpes simplex virus type 1 nucleocytoplasmic shuttling protein UL47 is mediated by an N-terminal arginine-rich domain that also functions as its nuclear localization signal. J Virol 81:2283–2296 [CrossRef]
    [Google Scholar]
  9. Eisfeld A. J., Turse S. E., Jackson S. A., Lerner E. C., Kinchington P. R. 2006; Phosphorylation of the varicella-zoster virus (VZV) major transcriptional regulatory protein IE62 by the VZV open reading frame 66 protein kinase. J Virol 80:1710–1723 [CrossRef]
    [Google Scholar]
  10. Frame M. C., Purves F. C., McGeoch D. J., Marsden H. S., Leader D. P. 1987; Identification of the herpes simplex virus protein kinase as the product of viral gene US3. J Gen Virol 68:2699–2704 [CrossRef]
    [Google Scholar]
  11. Geiss B. J., Cano G. L., Tavis J. E., Morrison L. A. 2004; Herpes simplex virus 2 VP22 phosphorylation induced by cellular and viral kinases does not influence intracellular localization. Virology 330:74–81 [CrossRef]
    [Google Scholar]
  12. Goshima F., Daikoku T., Yamada H., Oshima S., Tsurumi T., Nishiyama Y. 1998; Subcellular localization of the US3 protein kinase of herpes simplex virus type 2. Arch Virol 143:613–622 [CrossRef]
    [Google Scholar]
  13. Jones C., Chowdhury S. 2007; A review of the biology of bovine herpesvirus type 1 (BHV-1), its role as a cofactor in the bovine respiratory disease complex and development of improved vaccines. Anim Health Res Rev 8:187–205 [CrossRef]
    [Google Scholar]
  14. Kato A., Yamamoto M., Ohno T., Kodaira H., Nishiyama Y., Kawaguchi Y. 2005; Identification of proteins phosphorylated directly by the Us3 protein kinase encoded by herpes simplex virus 1. J Virol 79:9325–9331 [CrossRef]
    [Google Scholar]
  15. Kato A., Arii J., Shiratori I., Akashi H., Arase H., Kawaguchi Y. 2009; Herpes simplex virus 1 protein kinase Us3 phosphorylates viral envelope glycoprotein B and regulates its expression on the cell surface. J Virol 83:250–261 [CrossRef]
    [Google Scholar]
  16. Klupp B. G., Granzow H., Mettenleiter T. C. 2001; Effect of the pseudorabies virus US3 protein on nuclear membrane localization of the UL34 protein and virus egress from the nucleus. J Gen Virol 82:2363–2371
    [Google Scholar]
  17. Krek W., Maridor G., Nigg E. A. 1992; Casein kinase II is a predominantly nuclear enzyme. J Cell Biol 116:43–55 [CrossRef]
    [Google Scholar]
  18. LaBoissiere S., Trudel M., Simard C. 1992; Characterization and transcript mapping of a bovine herpesvirus type 1 gene encoding a polypeptide homologous to the herpes simplex virus type 1 major tegument proteins VP13/14. J Gen Virol 73:2941–2947 [CrossRef]
    [Google Scholar]
  19. Leach N., Bjerke S. L., Christensen D. K., Bouchard J. M., Mou F., Park R., Baines J., Haraguchi T., Roller R. J. 2007; Emerin is hyperphosphorylated and redistributed in herpes simplex virus type 1-infected cells in a manner dependent on both UL34 and US3. J Virol 81:10792–10803 [CrossRef]
    [Google Scholar]
  20. Leader D. P., Deana A. D., Marchiori F., Purves F. C., Pinna L. A. 1991; Further definition of the substrate specificity of the alpha-herpesvirus protein kinase and comparison with protein kinases A and C. Biochim Biophys Acta 1091:426–431 [CrossRef]
    [Google Scholar]
  21. Lemaster S., Roizman B. 1980; Herpes simplex virus phosphoproteins. II. Characterization of the virion protein kinase and of the polypeptides phosphorylated in the virion. J Virol 35:798–811
    [Google Scholar]
  22. Leopardi R., Van Sant C., Roizman B. 1997; The herpes simplex virus 1 protein kinase US3 is required for protection from apoptosis induced by the virus. Proc Natl Acad Sci U S A 94:7891–7896 [CrossRef]
    [Google Scholar]
  23. Malik P., Clements J. B. 2004; Protein kinase CK2 phosphorylation regulates the interaction of Kaposi's sarcoma-associated herpesvirus regulatory protein ORF57 with its multifunctional partner hnRNP K. Nucleic Acids Res 32:5553–5569 [CrossRef]
    [Google Scholar]
  24. Matsuzaki A., Yamauchi Y., Kato A., Goshima F., Kawaguchi Y., Yoshikawa T., Nishiyama Y. 2005; US3 protein kinase of herpes simplex virus type 2 is required for the stability of the UL46-encoded tegument protein and its association with virus particles. J Gen Virol 86:1979–1985 [CrossRef]
    [Google Scholar]
  25. Medina-Palazon C., Gruffat H., Mure F., Filhol O., Vingtdeux-Didier V., Drobecq H., Cochet C., Sergeant N., Sergeant A., Manet E. 2007; Protein kinase CK2 phosphorylation of EB2 regulates its function in the production of Epstein–Barr virus infectious viral particles. J Virol 81:11850–11860 [CrossRef]
    [Google Scholar]
  26. Meggio F., Pinna L. A. 2003; One-thousand-and-one substrates of protein kinase CK2?. FASEB J 17:349–368 [CrossRef]
    [Google Scholar]
  27. Meredith D. M., Lindsay J. A., Halliburton I. W., Whittaker G. R. 1991; Post-translational modification of the tegument proteins (VP13 and VP14) of herpes simplex virus type 1 by glycosylation and phosphorylation. J Gen Virol 72:2771–2775 [CrossRef]
    [Google Scholar]
  28. Misra V., Babiuk L. A., Darcel C. L. 1983; Analysis of bovine herpes virus-type 1 isolates by restriction endonuclease fingerprinting. Arch Virol 76:341–354 [CrossRef]
    [Google Scholar]
  29. Morrison E. E., Wang Y. F., Meredith D. M. 1998; Phosphorylation of structural components promotes dissociation of the herpes simplex virus type 1 tegument. J Virol 72:7108–7114
    [Google Scholar]
  30. Mou F., Forest T., Baines J. D. 2007; US3 of herpes simplex virus type 1 encodes a promiscuous protein kinase that phosphorylates and alters localization of lamin A/C in infected cells. J Virol 81:6459–6470 [CrossRef]
    [Google Scholar]
  31. Mou F., Wills E., Baines J. D. 2009; Phosphorylation of the UL31 protein of herpes simplex virus 1 by the US3-encoded kinase regulates localization of the nuclear envelopment complex and egress of nucleocapsids. J Virol 83:5181–5191 [CrossRef]
    [Google Scholar]
  32. Pagano M. A., Meggio F., Ruzzene M., Andrzejewska M., Kazimierczuk Z., Pinna L. A. 2004; 2-Dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole: a novel powerful and selective inhibitor of protein kinase CK2. Biochem Biophys Res Commun 321:1040–1044 [CrossRef]
    [Google Scholar]
  33. Perkins D. N., Pappin D. J., Creasy D. M., Cottrell J. S. 1999; Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20:3551–3567 [CrossRef]
    [Google Scholar]
  34. Pinna L. A. 2002; Protein kinase CK2: a challenge to canons. J Cell Sci 115:3873–3878 [CrossRef]
    [Google Scholar]
  35. Purves F. C., Deana A. D., Marchiori F., Leader D. P., Pinna L. A. 1986; The substrate specificity of the protein kinase induced in cells infected with herpesviruses: studies with synthetic substrates [corrected] indicate structural requirements distinct from other protein kinases. Biochim Biophys Acta 889:208–215 [CrossRef]
    [Google Scholar]
  36. Purves F. C., Longnecker R. M., Leader D. P., Roizman B. 1987; Herpes simplex virus 1 protein kinase is encoded by open reading frame US3 which is not essential for virus growth in cell culture. J Virol 61:2896–2901
    [Google Scholar]
  37. Rena G., Bain J., Elliott M., Cohen P. 2004; D4476, a cell-permeant inhibitor of CK1, suppresses the site-specific phosphorylation and nuclear exclusion of FOXO1a. EMBO Rep 5:60–65 [CrossRef]
    [Google Scholar]
  38. Russo G. L., Tosto M., Mupo A., Castellano I., Cuomo A., Tosti E. 2004; Biochemical and functional characterization of protein kinase CK2 in ascidian Ciona intestinalis oocytes at fertilization. Cloning and sequence analysis of cDNA for alpha and beta subunits. J Biol Chem 279:33012–33023 [CrossRef]
    [Google Scholar]
  39. Ryckman B. J., Roller R. J. 2004; Herpes simplex virus type 1 primary envelopment: UL34 protein modification and the US3–UL34 catalytic relationship. J Virol 78:399–412 [CrossRef]
    [Google Scholar]
  40. Shen W., Westgard E., Huang L., Ward M. D., Osborn J. L., Chau N. H., Collins L., Marcum B., Koach M. A. other authors 2008; Nuclear trafficking of the human cytomegalovirus pp71 (ppUL82) tegument protein. Virology 376:42–52 [CrossRef]
    [Google Scholar]
  41. Takashima Y., Tamura H., Xuan X., Otsuka H. 1999; Identification of the US3 gene product of BHV-1 as a protein kinase and characterization of BHV-1 mutants of the US3 gene. Virus Res 59:23–34 [CrossRef]
    [Google Scholar]
  42. Turin L., Russo S., Poli G. 1999; BHV-1: new molecular approaches to control a common and widespread infection. Mol Med 5:261–284
    [Google Scholar]
  43. van Drunen Littel-van den Hurk S., Garzon S., van den Hurk J. V., Babiuk L. A., Tijssen P. 1995; The role of the major tegument protein VP8 of bovine herpesvirus-1 in infection and immunity. Virology 206:413–425 [CrossRef]
    [Google Scholar]
  44. Verhagen J., Donnelly M., Elliott G. 2006; Characterization of a novel transferable CRM-1-independent nuclear export signal in a herpesvirus tegument protein that shuttles between the nucleus and cytoplasm. J Virol 80:10021–10035 [CrossRef]
    [Google Scholar]
  45. Wadd S., Bryant H., Filhol O., Scott J. E., Hsieh T. Y., Everett R. D., Clements J. B. 1999; The multifunctional herpes simplex virus IE63 protein interacts with heterogeneous ribonucleoprotein K and with casein kinase 2. J Biol Chem 274:28991–28998 [CrossRef]
    [Google Scholar]
  46. Wisner T. W., Wright C. C., Kato A., Kawaguchi Y., Mou F., Baines J. D., Roller R. J., Johnson D. C. 2009; Herpesvirus gB-induced fusion between the virion envelope and outer nuclear membrane during virus egress is regulated by the viral US3 kinase. J Virol 83:3115–3126 [CrossRef]
    [Google Scholar]
  47. Zaharevitz D. W., Gussio R., Leost M., Senderowicz A. M., Lahusen T., Kunick C., Meijer L., Sausville E. A. 1999; Discovery and initial characterization of the paullones, a novel class of small-molecule inhibitors of cyclin-dependent kinases. Cancer Res 59:2566–2569
    [Google Scholar]
  48. Zhang G., Stevens R., Leader D. P. 1990; The protein kinase encoded in the short unique region of pseudorabies virus: description of the gene and identification of its product in virions and in infected cells. J Gen Virol 71:1757–1765 [CrossRef]
    [Google Scholar]
  49. Zheng C., Brownlie R., Babiuk L. A., van Drunen Littel-van den Hurk S. 2004; Characterization of nuclear localization and export signals of the major tegument protein VP8 of bovine herpesvirus-1. Virology 324:327–339 [CrossRef]
    [Google Scholar]
  50. Zhu J., Brownlie R., Liu Q., Babiuk L. A., Potter A., Mutwiri G. K. 2009; Characterization of bovine Toll-like receptor 8: ligand specificity, signaling essential sites and dimerization. Mol Immunol 46:978–990 [CrossRef]
    [Google Scholar]
/content/journal/jgv/10.1099/vir.0.013532-0
Loading
/content/journal/jgv/10.1099/vir.0.013532-0
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