1887

Abstract

Human cytomegalovirus (HCMV) displays an exceptionally restricted host range in tissue culture with human fibroblasts being the principal fully permissive system. Nevertheless, immediate early (IE) proteins are expressed following infection of many non-permissive cell types of human, simian and murine origin, and viral origin-dependent DNA synthesis has been reconstituted by transfection of plasmids into Vero cells, a non-permissive line from African green monkey. We have examined the accumulation of HCMV strain AD169 DNA, and the replication of transfected HCMV origin-containing plasmids, in infected Vero and human embryonic kidney 293 cells, which were previously reported to express the major IE protein in a small proportion of infected cells but to be non-permissive for viral DNA synthesis. In Vero cells accumulation of origin-containing plasmid but not viral DNA occurred, whilst in 293 cells both DNAs accumulated. Immunofluorescence experiments indicated that following infection with 3 p.f.u. per cell, a small fraction of both cell types expressed the UL44 DNA replication protein. Neither cell line, however, supported the generation of infectious progeny virus. These results suggest that IE proteins expressed in Vero and 293 cells can induce the synthesis of early proteins capable of functioning in viral DNA replication, but there is a failure in later events on the pathway to infectious virus production. This provides further support for transfected Vero cells being a valid system in which to study HCMV DNA synthesis, and suggests that 293 cells may also prove useful in similar experiments.

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2003-03-01
2019-12-11
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References

  1. Abbotts, A. P., Preston, V. G., Hughes, M., Patel, A. H. & Stow, N. D. ( 2000; ). Interaction of the herpes simplex virus type 1 packaging protein UL15 with full length and deleted forms of the UL28 protein. J Gen Virol 81, 2999–3009.
    [Google Scholar]
  2. Anders, D. G. & McCue, L. A. ( 1996; ). The human cytomegalovirus genes and proteins required for DNA synthesis. Intervirology 39, 378–388.
    [Google Scholar]
  3. Anders, D. G., Kacica, M. A., Pari, G. & Punturieri, S. M. ( 1992; ). Boundaries and structure of human cytomegalovirus oriLyt, a complex origin for lytic-phase DNA replication. J Virol 66, 3373–3384.
    [Google Scholar]
  4. Brown, J. M., Kaneshima, H. & Mocarski, E. S. ( 1995; ). Dramatic interstrain differences in the replication of human cytomegalovirus in SCID-Hu mice. J Infect Dis 171, 1599–1603.[CrossRef]
    [Google Scholar]
  5. Bystrevskaya, V. B., Lobova, T. V., Smirnov, V. N., Makarova, N. E. & Kushch, A. A. ( 1997; ). Centrosome injury in cells infected with human cytomegalovirus. J Struct Biol 120, 52–60.[CrossRef]
    [Google Scholar]
  6. Chee, M. S., Bankier, A. T., Beck, S. & 12 other authors ( 1990; ). Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol 154, 125–169.
    [Google Scholar]
  7. Einhorn, L., Gadler, H. & Wahren, B. ( 1982; ). Adsorption of purified human cytomegalovirus and induction of early antigens in different cells. J Med Virol 10, 225–234.[CrossRef]
    [Google Scholar]
  8. Everett, R. D. ( 1984; ). Trans-activation of transcription by herpes virus products: requirement for two HSV-1 immediate-early polypeptides for maximum activity. EMBO J 3, 3135–3141.
    [Google Scholar]
  9. Fortunato, E. A. & Spector, D. H. ( 1999; ). Regulation of human cytomegalovirus gene expression. Adv Virus Res 54, 61–128.
    [Google Scholar]
  10. García-Ramírez, J. J., Ruchti, F., Huang, H., Simmen, K., Angulo, A. & Ghazal, P. ( 2001; ). Dominance of virus over host factors in cross-species activation of human cytomegalovirus early gene expression. J Virol 75, 26–35.[CrossRef]
    [Google Scholar]
  11. Hodge, P. D. & Stow, N. D. ( 2001; ). Effects of mutations within the herpes simplex virus type 1 DNA encapsidation signal on packaging efficiency. J Virol 75, 8977–8986.[CrossRef]
    [Google Scholar]
  12. Huang, E. S. ( 1975; ). Human cytomegalovirus. IV. Specific inhibition of virus-induced DNA polymerase activity and viral DNA replication by phosphonoacetic acid. J Virol 16, 1560–1565.
    [Google Scholar]
  13. Iskenderian, A. C., Huang, L., Reilly, A., Stenberg, R. M. & Anders, D. G. ( 1996; ). Four of eleven loci required for transient complementation of human cytomegalovirus DNA replication cooperate to activate expression of replication genes. J Virol 70, 383–392.
    [Google Scholar]
  14. LaFemina, R. & Hayward, G. S. ( 1986; ). Constitutive and retinoic acid-inducible expression of cytomegalovirus immediate-early genes in human teratocarcinoma cells. J Virol 58, 434–440.
    [Google Scholar]
  15. LaFemina, R. L. & Hayward, G. S. ( 1988; ). Differences in cell type-specific blocks to immediate early gene expression and DNA replication of human, simian and murine cytomegalovirus. J Gen Virol 69, 355–374.[CrossRef]
    [Google Scholar]
  16. Mocarski, E. S. & Tan Courcelle, C. ( 2001; ). Cytomegaloviruses and their replication. In Fields Virology, 4th edn, pp. 2629–2673. Edited by D. M. Knipe & P. M. Howley. Philadelphia: Lippincott Williams & Wilkins.
  17. Pari, G. S. & Anders, D. G. ( 1993; ). Eleven loci encoding trans-acting factors are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA replication. J Virol 67, 7126–7131.
    [Google Scholar]
  18. Pari, G. S., Kacica, M. A. & Anders, D. G. ( 1993; ). Open reading frames UL44, IRS1/TRS1, and UL36–38 are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA synthesis. J Virol 67, 2575–2582.
    [Google Scholar]
  19. Pizzorno, M. C., O'Hare, P., Sha, L., LaFemina, R. L. & Hayward, G. S. ( 1988; ). Trans-activation and autoregulation of gene expression by immediate-early region 2 gene products of human cytomegalovirus. Journal of Virology 62, 1167–1179.
    [Google Scholar]
  20. Sarisky, R. T. & Hayward, G. S. ( 1996; ). Evidence that the UL84 gene product of human cytomegalovirus is essential for promoting oriLyt-dependent DNA replication and formation of replication compartments in cotransfection assays. J Virol 70, 7398–7413.
    [Google Scholar]
  21. Sinzger, C., Schmidt, K., Knapp, J., Kahl, M., Beck, R., Waldman, J., Hebart, H., Einsele, H. & Jahn, G. ( 1999; ). Modification of human cytomegalovirus tropism through propagation in vitro is associated with changes in the viral genome. J Gen Virol 80, 2867–2877.
    [Google Scholar]
  22. Sinzger, C., Kahl, M., Laib, K., Klingel, K., Rieger, P., Plachter, B. & Jahn, G. ( 2000; ). Tropism of human cytomegalovirus for endothelial cells is determined by a post-entry step dependent on efficient translocation to the nucleus. J Gen Virol 81, 3021–3035.
    [Google Scholar]
  23. Slobbe van Drunen, M. E., Hendrickx, A. T., Vossen, R. C., Speel, E. J., van dam Mieras, M. C. & Bruggeman, C. A. ( 1998; ). Nuclear import as a barrier to infection of human umbilical vein endothelial cells by human cytomegalovirus strain AD169. Virus Res 56, 149–156.[CrossRef]
    [Google Scholar]
  24. St Jeor, S. C., Albrecht, T. B., Funk, F. D. & Rapp, F. ( 1974; ). Stimulation of cellular DNA synthesis by human cytomegalovirus. J Virol 13, 353–362.
    [Google Scholar]
  25. Stow, N. D. ( 2001; ). Packaging of genomic and amplicon DNA by the herpes simplex virus type 1 UL25 null mutant, KUL25 NS. J Virol 75, 10755–10765.[CrossRef]
    [Google Scholar]
  26. Stow, N. D. & Wilkie, N. M. ( 1976; ). An improved technique for obtaining enhanced infectivity with herpes simplex virus type 1 DNA. J Gen Virol 33, 447–458.[CrossRef]
    [Google Scholar]
  27. Stow, N. D., Hammarsten, O., Arbuckle, M. I. & Elias, P. ( 1993; ). Inhibition of herpes simplex virus type 1 DNA replication by mutant forms of the origin-binding protein. Virology 196, 413–418.[CrossRef]
    [Google Scholar]
  28. Zhu, Y., Huang, L. & Anders, D. G. ( 1998; ). Human cytomegalovirus oriLyt sequence requirements. J Virol 72, 4989–4996.
    [Google Scholar]
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