1887

Abstract

The alphaherpesvirus UL34 protein is necessary for the primary envelopment of intranuclear capsids at the inner leaflet of the nuclear membrane. In herpes simplex virus type 1, the UL34 protein is exclusively phosphorylated by the protein kinase encoded by the non-essential US3 gene. To investigate the effect of the pseudorabies virus (PrV) US3 product on the intracellular localization of the UL34 protein and on virus morphogenesis, PrV US3 deletion mutants were isolated and characterized. Immunofluorescence analyses demonstrated that in the absence of the US3 protein, the localization of the UL34 polypeptide to the nuclear membrane was not as pronounced as that seen with US3, although immunoelectron microscopy indicated the presence of the UL34 protein in both leaflets of the nuclear membrane. Ultrastructurally, an accumulation of enveloped virions in the perinuclear space in large invaginations of the inner nuclear membrane was observed, which were shown by immunoelectron microscopy to contain the UL34 protein, but not glycoproteins gB or gC. Thus, the US3 protein appears to be involved in the de-envelopment of perinuclear virions by fusion with the outer leaflet of the nuclear membrane. Surprisingly, no difference in the phosphorylation of the PrV UL34 protein was observed in the presence or absence of the US3 kinase. Therefore, the observed effects of the PrV US3 protein on the intracellular localization of the UL34 protein and on virus morphogenesis are probably not due to the phosphorylation of the UL34 protein by the US3 kinase.

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2001-10-01
2024-10-14
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References

  1. Brack A., Dijkstra J., Granzow H., Klupp B. G., Mettenleiter T. C. 1999; Inhibition of virion maturation by simultaneous deletion of glycoproteins E, I, and M of pseudorabies virus. Journal of Virology 73:5364–5372
    [Google Scholar]
  2. Dietz P., Klupp B. G., Fuchs W., Köllner B., Weiland E., Mettenleiter T. C. 2000; Pseudorabies virus glycoprotein K requires the UL20 gene product for processing. Journal of Virology 74:5083–5090
    [Google Scholar]
  3. Foster T. P., Kousoulas K. G. 1999; Genetic analysis of the role of herpes simplex virus type 1 glycoprotein K in infectious virus production and egress. Journal of Virology 73:8457–8468
    [Google Scholar]
  4. Graham F. L., van der Eb A. J. 1973; A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467
    [Google Scholar]
  5. Granzow H., Weiland F., Jöns A., Klupp B. G., Karger A., Mettenleiter T. C. 1997; Ultrastructural analysis of the replication cycle of pseudorabies virus in cell culture: a reassessment. Journal of Virology 71:2072–2082
    [Google Scholar]
  6. Granzow H., Klupp B. G., Fuchs W., Veits J., Osterrieder N., Mettenleiter T. C. 2001; Egress of alphaherpesviruses: comparative ultrastructural study. Journal of Virology 75:3675–3684
    [Google Scholar]
  7. Jöns A., Mettenleiter T. C. 1997; Green fluorescent protein expressed by recombinant pseudorabies virus as an in vivo marker for viral replication. Journal of Virological Methods 66:283–292
    [Google Scholar]
  8. Kaplan A. S., Vatter A. E. 1959; A comparison of herpes simplex and pseudorabies viruses. Virology 13:78–92
    [Google Scholar]
  9. Kimman T. G., de Wind N., de Bruin T., de Visser Y., Voermans J. 1994; Inactivation of glycoprotein gE and thymidine kinase or the US3-encoded protein kinase synergistically decreases in vivo replication of pseudorabies virus and the induction of protective immunity. Virology 205:511–518
    [Google Scholar]
  10. Klupp B. G., Baumeister J., Dietz P., Granzow H., Mettenleiter T. C. 1998; Pseudorabies virus glycoprotein gK is a virion structural component involved in virus release but is not required for entry. Journal of Virology 72:1949–1958
    [Google Scholar]
  11. Klupp B. G., Granzow H., Mettenleiter T. C. 2000; Primary envelopment of pseudorabies virus at the nuclear membrane requires the UL34 gene product. Journal of Virology 74:10063–10073
    [Google Scholar]
  12. Lukács N., Thiel H.-J., Mettenleiter T. C., Rziha H.-J. 1985; Demonstration of three major species of pseudorabies virus glycoproteins and identification of a disulfide-linked glycoprotein complex. Journal of Virology 53:166–173
    [Google Scholar]
  13. Peeters B., de Wind N., Hooisma M., Wagenaar F., Gielkens A., Moormann R. 1992; Pseudorabies virus envelope glycoproteins gp50 and gII are essential for virus penetration, but only gII is involved in membrane fusion. Journal of Virology 66:894–905
    [Google Scholar]
  14. 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 indicate structural requirements distinct from other protein kinases. Biochimica et Biophysica Acta 889:208–215
    [Google Scholar]
  15. 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. Journal of Virology 61:2896–2901
    [Google Scholar]
  16. Purves F. C., Spector D., Roizman B. 1991; The herpes simplex virus 1 protein kinase encoded by the US3 gene mediates posttranslational modification of the phosphoprotein encoded by the UL34 gene. Journal of Virology 65:5757–5764
    [Google Scholar]
  17. Purves F. C., Spector D., Roizman B. 1992; UL34, the target of the herpes simplex US3 protein kinase, is a membrane protein which in its unphosphorylated state associates with novel phosphoproteins. Journal of Virology 66:4295–4303
    [Google Scholar]
  18. Roller R., Zhou Y., Schnetzer R., Ferguson J., DeSalvo D. 2000; Herpes simplex virus type 1 UL34 gene product is required for viral envelopment. Journal of Virology 74:117–129
    [Google Scholar]
  19. van Zijl M., van der Gulden H., de Wind N., Gielkens A., Berns A. 1990; Identification of two genes in the unique short region of pseudorabies virus; comparison with herpes simplex virus and varicella-zoster virus. Journal of General Virology 71:1747–1755
    [Google Scholar]
  20. Wagenaar F., Pol J. M. A., Peeters B., Gielkens A. L. J., de Wind N., Kimman T. G. 1995; The US3-encoded protein kinase from pseudorabies virus affects egress of virions from the nucleus. Journal of General Virology 76:1851–1859
    [Google Scholar]
  21. Zhang G., Leader D. P. 1990; The structure of the pseudorabies virus genome at the end of the inverted repeat sequences proximal to the junction with the short unique region. Journal of General Virology 71:2433–2441
    [Google Scholar]
  22. 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. Journal of General Virology 71:1757–1765
    [Google Scholar]
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