1887

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Volume 76, Issue 6

Quiescent viral genomes in human fibroblasts after infection with herpes simplex virus type 1 Vmw65 mutants Free

Abstract

The development and utilization of a tissue culture system for the analysis of quiescent, nonreplicating herpes simplex virus type 1 (HSV-1) genomes is described. It was demonstrated previously that the HSV-1 Vmw65 mutant 1814, which is impaired for immediate early (IE) transcription, was retained for many days in human fetal lung (HFL) fibroblasts in a quiescent ‘latent’ state. Molecular analysis of the viral genome was not possible, however, due to residual expression of IE proteins and consequent cytotoxicity at high m.o.i. In the study reported here, IE transcription was reduced further by pretreatment of cells with interferon-α (IFN-α) and by the the use of mutant 1820, a derivative of 1814 in which the Vmw110 promoter was replaced by the Moloney murine leukaemia virus (Momulv) enhancer. The Momulv enhancer was not expressed under IE conditions; thus 1820 was more impaired for replication than 1814 and behaved as if deficient for both Vmw65 and Vmw110. In cells pretreated with IFN-α and subsequently infected with 1820 cytotoxicity was overcome, enabling a tissue culture system to be developed in which all cells stably retained at least one quiescent viral genome. To assist the analysis of gene expression, 1820 was further modified by insertion of the gene controlled by the human cytomegalovirus enhancer (mutant 1883) or the HSV-1 immediate early Vmw110 promoter (1884). Expression of -galactosidase was not detected after infection of IFN--pretreated cells with 1883 or 1884 but could be induced in almost all cells containing a viral genome, by superinfection of cultures. 1820-derived viruses were retained for at least 9 days and were not reactivated by subculture of cells. A regular arrangement of nucleosomes, as found in cellular chromatin, was not detected on the viral genome at the thymidine kinase locus. The non-linear genome was a template for reactivation with no requirement for prior conversion to a linear form. A small number of remaining linear genomes resulted from incomplete uncoating of input virus.

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© Journal of General Virology 1995
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1995-06-01
2024-04-20
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References

  1. Ace C. I., Dalrymple M. A., Ramsay F. H., Preston V. G., Preston C. M. 1988; Mutational analysis of the herpes simplex virus type 1 tram-inducing factor Vmw65. Journal of General Virology 69:2595–2605
     [Google Scholar]
  2. Ace C. I., McKee T. A., Ryan J. M., Cameron J. M., Preston C. M. 1989; Construction and characterization of a herpes simplex virus type 1 mutant unable to transinduce immediate-early gene expression. Journal of Virology 63:2260–2269
     [Google Scholar]
  3. Brennan M. B., Stark G. R. 1983; Interferon pretreatment inhibits simian virus 40 infections by blocking the onset of early transcription. Cell 33:811–816
     [Google Scholar]
  4. Cai W., Schaffer P. A. 1991; A cellular function can enhance gene expression and plating efficiency of a mutant defective in the gene for ICP0, a transactivating protein of herpes simplex virus type 1. Journal of Virology 65:4078–4090
     [Google Scholar]
  5. Cai W., Schaffer P. A. 1992; Herpes simplex virus type 1 ICPO regulates expression of immediate-early, early and late genes in productively infected cells. Journal of Virology 66:2904–2915
     [Google Scholar]
  6. 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]
  7. Chen J., Silverstein S. 1992; Herpes simplex viruses with mutations in the gene encoding ICPO are defective in gene expression. Journal of Virology 66:2916–2927
     [Google Scholar]
  8. Cheung P., Banfield B. W., Tufaro F. 1991; Brefeldin A arrests the maturation and progress of herpes simplex virus particles during infection. Journal of Virology 65:1893–1904
     [Google Scholar]
  9. Croen K. D., Ostrove J. M., Dragovic L. J., Straus S. E. 1987; Latent herpes simplex virus in human trigeminal ganglia: detection of an immediate-early gene ‘antisense’ transcript by in situ hybridization. New England Journal of Medicine 317:1427–1432
     [Google Scholar]
  10. Daksis J. I., Preston C. M. 1992; Herpes simplex immediate early gene expression in the absence of transinduction by Vmw65 varies during the cell cycle. Virology 189:196–202
     [Google Scholar]
  11. Davison M.-J., Preston V. G., McGeoch D. J. 1984; Determination of the sequence alteration in the DNA of the herpes simplex virus type 1 temperature-sensitive mutant ts K. Journal of General Virology 65:859–863
     [Google Scholar]
  12. Deshmane S. L., Fraser N. W. 1989; During latency, herpes simplex virus type 1 DNA is associated with nucleosomes in a chromatin structure. Journal of Virology 63:943–947
     [Google Scholar]
  13. DeStasio R. P., Taylor M. W. 1990; Specific effect of interferon on the herpes simplex virus type 1 transactivation event. Journal of Virology 64:2588–2593
     [Google Scholar]
  14. Dixon R. A. F., Schaffer P. A. 1980; Fine-structure mapping and functional analysis of temperature-sensitive mutants in the gene encoding the herpes simplex virus type 1 immediate early protein VP175. Journal of Virology 36:189–203
     [Google Scholar]
  15. Ecob-Prince M. S., Hassan K. 1994; Reactivation of latent herpes simplex virus from explanted dorsal root ganglia. Journal of General Virology 75:2017–2028
     [Google Scholar]
  16. Ecob-Prince M. S., Preston C. M., Rixon F. J., Hassan K., Kennedy P. G. E. 1993; Neurons containing latency-associated transcripts are numerous and widespread in dorsal root ganglia following footpad inoculation of mice with herpes simplex virus type 1 mutant in 1814. Journal of General Virology 74:985–994
     [Google Scholar]
  17. Efstathiou S., Minson A. C., Field H. J., Anderson J. R., Wildy P. 1986; Detection of herpes simplex virus-specific DNA sequences in latently infected mice and humans. Journal of Virology 57:446–455
     [Google Scholar]
  18. Everett R. D. 1984; Transactivation of transcription by herpes virus products: requirement for two HSV-1 immediate-early polypeptides for maximum activity. EMBO Journal 3:3135–3141
     [Google Scholar]
  19. Everett R. D. 1987; The regulation of transcription of viral and cellular genes by herpesvirus immediate-early gene products. Anticancer Research 7:589–604
     [Google Scholar]
  20. Everett R. D. 1989; Construction and characterisation of herpes simplex virus type 1 mutants with defined lesions in immediate early gene 1. Journal of General Virology 70:1185–1202
     [Google Scholar]
  21. Feinberg A. P., Vogelstein B. 1983; A technique for radiolabelling restriction endonuclease fragments to a high specific activity. Analytical Biochemistry 132:6–13
     [Google Scholar]
  22. Fraser N. W., Block T. M., Spivack J. G. 1992; The latency-associated transcripts of herpes simplex virus: RNA in search of a function. Virology 191:1–8
     [Google Scholar]
  23. Greaves R., O’Hare P. 1990; Structural requirements in the herpes simplex virus type 1 transactivator Vmw65 for interaction with the cellular octamer-binding protein and target TAATGARAT sequences. Journal of Virology 64:2716–2724
     [Google Scholar]
  24. Harris R. A., Preston C. M. 1991; Establishment of latency in vitro by the herpes simplex virus type 1 mutant in 1814. Journal of General Virology 72:907–913
     [Google Scholar]
  25. Hayes S., O’Hare P. 1993; Mapping of a major surface-exposed site in herpes simplex virus protein Vmw65 to a region of direct interaction in a transcription complex assembly. Journal of Virology 67:852–862
     [Google Scholar]
  26. 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]
  27. Jacob R. J., Roizman B. 1977; Anatomy of herpes simplex virus DNA. VII. Properties of the replicating DNA. Journal of Virology 23:394–411
     [Google Scholar]
  28. Johnson P. A., Miyanohara A., Levine F., Cahill T., Friedmann T. 1992; Cytotoxicity of a replication defective mutant of herpes simplex virus type 1. Journal of Virology 66:2952–2965
     [Google Scholar]
  29. Johnson P. A., Wang M. J., Friedmann T. 1994; Improved cell survival by the reduction of immediate-early gene expression in replication-defective mutants of herpes simplex virus type 1 but not by mutation of the virion host shutoff function. Journal of Virology 68:6347–6362
     [Google Scholar]
  30. Katan M., Haigh A., Verrijzer P. C., Van Der Vliet P. C., O’Hare P. 1990; Characterization of a cellular factor which interacts functionally with oct-1 in the assembly of a multicomponent transcription complex. Nucleic Acids Research 18:6871–6880
     [Google Scholar]
  31. Kemp L. M., Dent C. L., Latchman D. S. 1990; Octamer motif mediates transcriptional repression of HSV immediate-early genes and octamer-containing cellular promoters in neuronal cells. Neuron 4:215–222
     [Google Scholar]
  32. Kosz-Vnenchak M., Jacobsen J., Coen D. M., Knife D. M. 1993; Evidence for a novel regulatory pathway for herpes simplex virus gene expression in trigeminal ganglion neurons. Journal of Virology 67:5383–5393
     [Google Scholar]
  33. Kristie T. M., Roizman B. 1988; Differentiation and DNA contact points of the host proteins binding at the as site for virion-mediated induction of herpes simplex virus 1 a genes. Journal of Virology 62:1145–1157
     [Google Scholar]
  34. Kristie T. M., Sharp P. A. 1990; Interactions of the oct-1 POU subdomains with specific DNA sequences and with the HSV α-trans-activator protein. Genes & Development 4:2383–2396
     [Google Scholar]
  35. Lang J. C., Wilkie N. M., Spandidos D. A. 1983; Characterization of eukaryotic transcriptional control signals by assay of herpes simplex virus type 1 thymidine kinase. Journal of General Virology 64:2679–2696
     [Google Scholar]
  36. Leinbach S. S., Summers W. C. 1980; The structure of herpes simplex virus type 1 DNA as probed by micrococcal nuclease digestion. Journal of General Virology 51:45–59
     [Google Scholar]
  37. Lillycrop K. A., Dent C. L., Wheatley S. C., Beech N. M., Ninkina N. N., Wood J. N., Latchman D. S. 1991; The octamer-binding protein oct-2 represses HSV immediate-early genes in cell lines derived from latently infectable sensory neurons. Neuron 7:381–390
     [Google Scholar]
  38. Lillycrop K. A., Howard M. K., Estridge J. K., Latchman D. S. 1994; Inhibition of herpes simplex virus infection by ectopic expression of neuronal splice variants of the oct-2 transcription factor. Nucleic Acids Research 22:815–820
     [Google Scholar]
  39. McFarlane M., Daksis J. I., Preston C. M. 1992; Hexa-methylene bisacetamide stimulates herpes simplex virus immediate early gene expression in the absence of trans-induction by Vmw65. Journal of General Virology 73:285–292
     [Google Scholar]
  40. McKnight J. L. C., Kristie T. M., Roizman B. 1987; Binding of the virion protein mediating a gene induction in herpes simplex virus 1-infected cells to its cis site requires cellular proteins. Proceedings of the National Academy of Sciences, USA 84:7061–7065
     [Google Scholar]
  41. Mittnach S., Straub P., Kirchner H., Jacobsen H. 1988; Interferon treatment inhibits onset of herpes simplex virus immediate-early transcription. Virology 164:201–210
     [Google Scholar]
  42. Muggeridge M. I., Fraser N. W. 1986; Chromosomal organization of the herpes simplex virus genome during acute infection of the mouse central nervous system. Journal of Virology 59:764–767
     [Google Scholar]
  43. Oberman F., Panet A. 1989; Characterization of the early steps of herpes simplex virus replication in interferon-treated human cells. Journal of Interferon Research 9:563–571
     [Google Scholar]
  44. O’Hare P., Goding C. R. 1988; Herpes simplex virus regulatory elements and the immunoglobulin octamer domain bind a common factor and are both targets for virion transactivation. Celt 54:435–445
     [Google Scholar]
  45. O’Hare P., Hayward G. S. 1985; Three trans-acting regulatory proteins of herpes simplex virus modulate immediate-early gene expression in a pathway involving positive and negative feedback regulation. Journal of Virology 56:723–733
     [Google Scholar]
  46. Perry L. J., McGeoch D. J. 1988; The DNA sequences of the long repeat region and adjoining parts of the long unique region in the genome of herpes simplex virus type 1. Journal of General Virology 69:2831–2846
     [Google Scholar]
  47. Poffenberger K. L., Roizman B. 1985; A noninverting genome of a viable herpes simplex virus 1: presence of head-to-tail linkages in packaged genomes and requirements for circularization after infection. Journal of Virology 53:587–595
     [Google Scholar]
  48. Post L. E., Mackem S., Roizman B. 1981; Regulation of α genes of herpes simplex virus: expression of chimeric genes produced by fusion of thymidine kinase with a gene promoters. Cell 24:555–565
     [Google Scholar]
  49. Preston C. M. 1979a; Abnormal properties of an immediate early polypeptide in cells infected with the herpes simplex virus type 1 mutant ts K. Journal of Virology 32:357–369
     [Google Scholar]
  50. Preston C. M. 1979b; Control of herpes simplex virus type 1 mRNA synthesis in cells infected with wild-type virus or the temperature sensitive mutant ts K. Journal of Virology 29:275–284
     [Google Scholar]
  51. Preston C. M., Frame M. C., Campbell M. E. M. 1988; A complex formed between cell components and an HSV structural polypeptide binds to a viral immediate early gene regulatory sequence. Cell 52:425–434
     [Google Scholar]
  52. Ralph W. M., Cabatingan M. S., Schaffer P. A. 1994; Induction of herpes simplex virus immediate-early gene expression by a cellular activity expressed in vero and NB41A3 cells after growth arrest-release. Journal of Virology 68:6871–6882
     [Google Scholar]
  53. Rixon F. J., McLauchlan J. 1990; Insertion of DNA sequences at a unique restriction enzyme site engineered for vector purposes into the genome of herpes simplex virus type 1. Journal of General Virology 71:2931–2939
     [Google Scholar]
  54. Rock D. L., Fraser N. W. 1983; Detection of HSV-1 genome in central nervous system of latently infected mice. Nature 302:523–525
     [Google Scholar]
  55. Roizman B., Sears A. E. 1987; An inquiry into the mechanisms of herpes simplex virus latency. Annual Review of Microbiology 41:543–571
     [Google Scholar]
  56. Sacks W. R., Greene C. C., Aschman D. P., Schaffer P. A. 1985; Herpes simplex virus type 1 ICP27 is an essential regulatory protein. Journal of Virology 55:796–805
     [Google Scholar]
  57. Sacks W. R., Schaffer P. A. 1987; Deletion mutants in the gene encoding the herpes simplex virus type 1 immediate-early protein ICPO exhibit impaired growth in culture. Journal of Virology 61:829–839
     [Google Scholar]
  58. Sadowski I., Ma J., Triezenberg S. J., Ptashne M. 1988; GAL4-VP16 is an unusually potent transcriptional activator. Nature 335:563–564
     [Google Scholar]
  59. Sandri-Goldin R. M., Mendoza G. E. 1992; A herpesvirus regulatory protein appears to act post-transcriptionally by affecting mRNA processing. Genes & Development 6:848–863
     [Google Scholar]
  60. Sears A. E., Hukkanen V., Labow M. A., Levine A. J., Roizman B. 1991; Expression of herpes simplex virus 1 α transinducing factor (VP16) does not induce reactivation of latent virus or prevent the establishment of latency in mice. Journal of Virology 65:2929–2935
     [Google Scholar]
  61. Simmons A., Slobedman B., Speck P., Arthur J., Efstathiou S. 1992; Two patterns of persistence of herpes simplex virus DNA sequences in the nervous systems of latently infected mice. Journal of General Virology 73:1287–1291
     [Google Scholar]
  62. Slobedman B., Efstathiou S., Simmons A. 1994; Quantitative analysis of herpes simplex virus DNA and transcriptional activity in ganglia of mice latently infected with wild-type and thymidine kinase-deficient viral strains. Journal of General Virology 75:2469–2474
     [Google Scholar]
  63. Smibert C. A., Popova B., Xiao P., Capone J. P., Smiley J. R. 1994; Herpes simplex virus VP16 forms a complex with the virion host shutoff protein vhs. Journal of Virology 68:2339–2346
     [Google Scholar]
  64. Spivack J. G., Fraser N. W. 1987; Detection of herpes simplex virus type 1 transcripts during latent infection in mice. Journal of Virology 61:3841–3847
     [Google Scholar]
  65. Steiner I., Spivack J. G., Deshmane S. L., Ace C. I., Preston C. M., Fraser N. W. 1990; A herpes simplex virus type 1 mutant containing a non-transinducing Vmw65 protein establishes latent infection in vivo in the absence of viral replication and reactivates efficiently from explanted trigeminal ganglia. Journal of Virology 64:1630–1638
     [Google Scholar]
  66. Stern S., Tanaka M., Herr W. 1989; The oct-1 homeodomain directs formation of a multiprotein-DNA complex with the HSV transactivator VP16. Nature 341:624–630
     [Google Scholar]
  67. Stevens J. G., Wagner E. K., Devi-Rao G. B., Cook M. L., Feldman L. 1987; RNA complementary to a herpesvirus alpha gene mRNA is predominant in latently infected neurons. Science 235:1056–1059
     [Google Scholar]
  68. Stinski M. F., Roehr T. J. 1985; Activation of the major immediate early gene of human cytomegalovirus by cis-acting elements in the promoter-regulatory sequence and by virus-specific trans-acting components. Journal of Virology 55:431–441
     [Google Scholar]
  69. Stow N. D., Stow E. C. 1986; Isolation and characterization of a herpes simplex virus type 1 mutant containing a deletion within the gene encoding the immediate early polypeptide Vmw110. Journal of General Virology 67:2571–2585
     [Google Scholar]
  70. Stow E. C., Stow N. D. 1989; Complementation of a herpes simplex virus type 1 Vmw110 mutant by human cytomegalovirus. Journal of General Virology 70:695–704
     [Google Scholar]
  71. Triezenberg S. J., Kingsbury R. C., McKnight S. L. 1988; Functional dissection of VP16, the trans-activator of herpes simplex virus immediate early gene expression. Genes & Development 2:718–729
     [Google Scholar]
  72. Valyi-Nagy T., Deshmane S. L., Spivack J. G., Steiner I., Ace C. I., Preston C. M., Fraser N. W. 1991; Investigation of herpes simplex virus type 1 (HSV-1) gene expression and DNA synthesis during the establishment of latent infection by an HSV-1 mutant, in 1814, that does not replicate in mouse trigeminal ganglia. Journal of Virology 72:641–649
     [Google Scholar]
  73. Watson R. J., Clements J. B. 1980; A herpes simplex virus type 1 function required for early and late virus RNA synthesis. Nature 285:329–330
     [Google Scholar]
  74. Weinheimer S. P., Boyd B. A., Durham S. K., Resnick J. L., O’Boyle D. R. 1992; Deletion of the VP16 open reading frame of herpes simplex virus type 1. Journal of Virology 66:258–269
     [Google Scholar]
  75. Wigdahl B. L., Scheck A. C., Declercq E., Rapp F. 1982; High efficiency latency and activation of herpes simplex virus in human cells. Science 217:1145–1146
     [Google Scholar]
  76. Wigdahl B. L., Scheck A. C., Ziegler R. J., Declercq E., Rapp F. 1984; Analysis of the herpes simplex virus genome during latency in human diploid fibroblasts and rat sensory neurons. Journal of Virology 49:205–213
     [Google Scholar]
  77. Wilkie N. M., Clements J. B., Boll W., Mantei N., Lonsdale D., Weissman C. 1979; Hybrid plasmids containing an active thymidine kinase gene of herpes simplex virus. Nucleic Acids Research 7:859–877
     [Google Scholar]
  78. Wilson A. C., Lamarco K., Peterson M. G., Herr W. 1993; The VP 16 accessory protein HCF is a family of polypeptides processed from a large precursor protein. Cell 74:115–125
     [Google Scholar]
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Quiescent viral genomes in human fibroblasts after infection with herpes simplex virus type 1 Vmw65 mutants
J. Gen. Virol. 76, 1417 (1995); https://doi.org/10.1099/0022-1317-76-6-1417
/content/journal/jgv/10.1099/0022-1317-76-6-1417
/content/journal/jgv/10.1099/0022-1317-76-6-1417
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