1887

Journal of General Virology header logo

Volume 79, Issue 10

Differences in the intracellular localization and fate of herpes simplex virus tegument proteins early in the infection of Vero cells. Free

Abstract

The fate of herpes simplex virus 1 (HSV-1) tegument proteins during infection in Vero cells was investigated immunochemically. Input virion-associated VP13/14 and VP16 localized to the nucleus early in infection, while VP1/2 localized to the nuclear envelope of the cell and VP22 could not be detected using monoclonal antibody P43. Western blotting suggested that virion-associated VP13/14, VP16 and VP22 were stable in infected cells whereas VP1/2 appeared to be processed or modified. Further studies showed that P43 recognized a phosphorylation-sensitive epitope in VP22 and suggested that virion-associated VP22 was phos- phorylated upon entry to the cell. VP13/14 and VP16 were easily extracted from cells early in infection whereas VP22 was largely insoluble. Phosphatase treatment of soluble extracts caused a shift in the molecular mass of VP16 showing it was phosphorylated. As infection progressed VP16 was observed in discrete nuclear compartments where it co-localized with ICP8 and the capsid-associated protein VP22a. VP13/14 was also observed in the nucleus. P43 immunostaining appeared around 6 h post-infection as punctate nuclear foci which often localized to the edge of VP16-immunoreactive areas. Punctate P43 cytoplasmic staining appeared around 12 h post-infection. By 18 h the nuclear pattern had disappeared and an extensive cytoplasmic stain was observed which closely overlapped that of other tegument proteins. On the basis of these data we suggest that virion-associated VP22 is phosphorylated upon entry of the virus into the cell and that unphosphorylated VP22, which is preferentially recognized by P43, becomes available later in infection, initially in the nucleus, for packaging into virions.

  • Published Online:
© Society for General Microbiology 1998
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-79-10-2517
1998-10-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/79/10/9780059.html?itemId=/content/journal/jgv/10.1099/0022-1317-79-10-2517&mimeType=html&fmt=ahah

References

  1. Batterson W., Furlong D., Roizman B. 1983; Molecular genetics of herpes simplex virus. VIII. Further characterization of a temperaturesensitive mutant defective in the release of viral DNA and in other stages of the viral reproductive cycle. Journal of Virology 45:397–407
     [Google Scholar]
  2. Campbell M. E. M., Palfreyman J. W., Preston C. M. 1984; Identification of herpes simplex virus DNA sequences which encode a trans-acting polypeptide responsible for stimulation of immediate-early transcription. Journal of Molecular Biology 180:1–19
     [Google Scholar]
  3. Coulter L. J., Moss H. M. W., Lang J., McGeoch D. J. 1993; A mutant of herpes simplex virus type 1 in which the UL13 protein kinase gene is disrupted. Journal of General Virology 74:387–395
     [Google Scholar]
  4. Elliott G. D., Meredith D. M. 1992; The herpes simplex virus type 1 tegument protein VP22 is encoded by gene UL49. Journal of General Virology 73:723–726
     [Google Scholar]
  5. Elliott G., O’Reilly D., O’Hare P. 1996; Phosphorylation of the herpes simplex virus type 1 tegument protein VP22. Virology 226:140–145
     [Google Scholar]
  6. Gibson W., Roizman B. 1974; Proteins specified by herpes simplex virus. χ. Staining and radiolabelling properties of B capsid and virion proteins in polyacrylamide gels. Journal of Virology 13:155–165
     [Google Scholar]
  7. Heine J. W., Honess R. W., Cassai E., Roizman B. 1974; Proteins specified by herpes simplex virus. XII. The virion polypeptides of type 1 strains. Journal of Virology 14:640–651
     [Google Scholar]
  8. Honess R. W., Roizman B. 1973; Proteins specified by herpes simplex virus. XI. Identification and relative molar rates of synthesis of structural and non-structural herpesvirus polypeptides in the infected cell. Journal of Virology 12:640–651
     [Google Scholar]
  9. Knipe D. M., Batterson W., Nosal C., Roizman B., Buchan A. 1981; Molecular genetics of herpes simplex virus. VI. Characterization of a temperature-sensitive mutant defective in the expression of all early viral gene products. Journal of Virology 38:539–547
     [Google Scholar]
  10. Knipe D. M., Senechek D., Rice S. A., Smith J. L. 1987; Stages in the nuclear association of the herpes simplex virus transcriptional activator protein ICP4. Journal of Virology 61:276–284
     [Google Scholar]
  11. Knopf K.-W., Kaerner H. C. 1980; Virus-specific basic phospho- proteins associated with herpes simplex virus type 1 (HSV-1) particles and the chromatin of HSV-1-infected cells. Journal of General Virology 46:405–415
     [Google Scholar]
  12. Liptak L. M., Uprichard S. L., Knipe D. M. 1996; Functional order of assembly of herpes simplex virus DNA replication proteins into prereplicative site structures. Journal of Virology 70:1759–1767
     [Google Scholar]
  13. McKnight J. L. C., Pellet P. E., Jenkins F. J., Roizman B. 1987; Characterization and nucleotide sequence of two herpes simplex virus 1 genes whose products modulate a-frans-inducing factor-dependent activation of α genes. Journal of Virology 61:992–1001
     [Google Scholar]
  14. McLean G., Rixon F., Langeland N., Haarr L., Marsden H. 1990; Identification and characterization of the virion protein products of herpes simplex virus type 1 gene UL47. Journal of General Virology 71:2953–2960
     [Google Scholar]
  15. McNabb D. S., Courtney R. J. 1992a; Analysis of the UL36 open reading frame encoding the large tegument protein (ICP1/2) of herpes simplex virus type 1. Journal of Virology 66:7581–7584
     [Google Scholar]
  16. McNabb D. S., Courtney R. J. 1992b; Characterization of the large tegument protein (ICP1/2) of herpes simplex virus type 1. Virology 190:221–232
     [Google Scholar]
  17. 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. Journal of General Virology 72:2771–2775
     [Google Scholar]
  18. Morrison E. E., Wang Y.-F., Meredith D. M. 1998; Phosphorylation of structural components promotes the dissociation of the HSV-1 tegument. Journal of Virology in press
    [Google Scholar]
  19. O’Reilly D., Hanscombe O., O’Hare P. 1997; A single serine residue at position 375 of VP16 is critical for complex assembly with Oct- 1 and HCF and is a target of phosphorylation by casein kinase II. EMBO Journal 16:2420–2430
     [Google Scholar]
  20. Phelan A., Clements J. B. 1997; Functional domains within the nucleus of a cell infected with HSV-1. Reviews in Medical Virology 7:229–237
     [Google Scholar]
  21. Pinard M.-F., Simard R., Bibor-Hardy V. 1987; DNA-binding proteins of herpes simplex virus type 1-infected BHK cell nuclear matrices. Journal of General Virology 68:727–735
     [Google Scholar]
  22. Quinlan M., Chen L.-B., Knipe D. 1984; The intranuclear location of a herpes simplex virus DNA-binding protein is determined by the status of viral DNA replication. Cell 36:857–868
     [Google Scholar]
  23. Randall R. E., Dinwoodie N. 1986; Intranuclear localization of herpes simplex virus immediate-early and delayed-early proteins: evidence that ICP 4 is associated with progeny virus DNA. Journal of General Virology 67:2163–2177
     [Google Scholar]
  24. Sodeik B., Ebersold M. W., Helenius A. 1997; Microtubule- mediated transport of incoming herpes simplex virus type 1 capsids to the nucleus. Journal of Cell Biology 130:1007–1022
     [Google Scholar]
  25. Spear P. G., Roizman B. 1972; Proteins specified by herpes simplex virus. V. Purification and structural proteins of the herpes virion. Journal of Virology 9:143–159
     [Google Scholar]
  26. Stevenson A. J., Morrison E. E., Chaudhari R., Yang C.-C., Meredith D. M. 1997; Processing and intracellular localization of the herpes simplex virus type 1 proteinase. Journal of General Virology 78:671–675
     [Google Scholar]
  27. Ward P. L., Ogle W. O., Roizman B. 1996; Assemblons : nuclear structures defined by aggregation of immature capsids and some tegument proteins of herpes simplex virus 1. Journal of Virology 70:4623–4631
     [Google Scholar]
  28. Whittaker G. R., Riggio M. P., Halliburton I. W., Killington R. A., Allen G. P., Meredith D. M. 1991; Antig enic and protein sequence homology between VP13/14, a herpes simplex type 1 tegument protein, and gp10, a glycoprotein of equine herpesvirus 1 and 4. Journal of Virology 65:2320–2326
     [Google Scholar]
  29. Wilcock D., Lane D. P. 1991; Localization of p53, retinoblastoma and host replication proteins at sites of viral replication in herpes-infected cells. Nature 349:429–431
     [Google Scholar]
  30. Yao F., Courtney R. J. 1989; A major transcriptional regulatory protein (ICP4) of herpes simplex virus type 1 is associated with purified virions. Journal of Virology 63:3338–3344
     [Google Scholar]
  31. Zhang Y., Sirko D. A., McKnight J. L. 1991; Role of herpes simplex virus type 1 UL46 and UL47 in αTIF-mediated transcriptional induction: characterization of three viral deletion mutants. Journal of Virology 65:829–841
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-79-10-2517
Loading
Differences in the intracellular localization and fate of herpes simplex virus tegument proteins early in the infection of Vero cells.
J. Gen. Virol. 79, 2517 (1998); https://doi.org/10.1099/0022-1317-79-10-2517
/content/journal/jgv/10.1099/0022-1317-79-10-2517
/content/journal/jgv/10.1099/0022-1317-79-10-2517
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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