1887

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Volume 68, Issue 11

Resistance to Methylation of the Human Cytomegalovirus Immediate Early Enhancer in a Model for Virus Latency and Reactivation Free

Abstract

Summary

Rat-9G cells carry several stably integrated copies of the major immediate early (IE) transcription unit of the human cytomegalovirus (HCMV). In these cells IE expression is repressed but inducible. In this report we describe the DNA methylation status of II, I and II sites within the IE gene, determined at different passage levels. Most, if not all, of the resident IE genes were progressively methylated in a similar fashion. This resulted in DNA methylation patterns in which sites surrounding the IE upstream region were preferentially methylated to a high degree. In contrast, sites within the 19 bp IE enhancer elements were markedly under-methylated. This particular DNA methylation pattern probably resulted from differences in DNA methylation rates, sites within the IE enhancer being methylated at only a very low rate. Methylation of the IE genes did not affect their inducibility, which might be related to the very low methylation level of the IE enhancer.

  • Received:
  • Accepted:
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Keyword(s): HCMV , immediate early enhancer and methylation
© Society for General Microbiology 1987
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1987-11-01
2024-04-25
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References

  1. Akrigg A., Wilkinson G. W. G., Oram J. D. 1985; The structure of the major immediate early gene of human cytomegalovirus. Virus Research 2:107–121
     [Google Scholar]
  2. Ball D. J., Gross D. S., Garrard T. 1983; 5-Methylcytosine is localized in nucleosomes that contain histone HI. Proceedings of the National Academy of Sciences, U.S.A 80:5490–5494
     [Google Scholar]
  3. Bird A. P. 1986; CpG rich islands and the function of DNA methylation. Nature; London: 321209–213
     [Google Scholar]
  4. Blanton R. A., Tevethia M. J. 1981; Immunoprecipitation of virus specific immediate early and early polypeptides from cells lytically infected with human cytomegalovirus strain AD169. Virology 112:262–273
     [Google Scholar]
  5. Bolden A. H., Nalin C. M., Ward C. A., Poonian M. S., Mccomas W. W., Weissbach A. 1985; DNA methylation: sequences flanking CG pairs modulate the specificity of the human DNA methylase. Nucleic Acids Research 13:3479–3494
     [Google Scholar]
  6. Bolden A. H., Nalin C. M., Ward C. A., Poonian M. S., Weissbach A. 1986; Primary DNA sequence determines sites of maintenance and de novo methylation by mammalian DNA methyltransferases. Molecular and Cellular Biology 6:1135–1140
     [Google Scholar]
  7. Boom R., Geelen J. L., Sol C. J., Raap A. K., Minnaar R. P., Klaver B. P., Van Der Noordaa J. 1986; Establishment of a rat cell line inducible for the expression of human cytomegalovirus immediate early gene products by protein synthesis inhibition. Journal of Virology 58:851–859
     [Google Scholar]
  8. Boshart M., Weber F., Jahn G., Dorsch-Häsler K., Fleckenstein B., Schaffner W. 1985; A Very Strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell 41:521–530
     [Google Scholar]
  9. Cameron J. M., Preston C. 1981; Comparison of the immediate early polypeptides of human cytomegalovirus isolates. Journal of General Virology 54:421–424
     [Google Scholar]
  10. Cooper D. N. 1983; Eukaryotic DNA methylation. Human Genetics 64:315–333
     [Google Scholar]
  11. Demarchi J. M. 1981; Human cytomegalovirus DNA: restriction enzyme cleavage maps and locations for immediate early, early and late RNAs. Virology 114:23–28
     [Google Scholar]
  12. Doereler W. 1981; DNA methylation - a regulatory signal in eukaryotic gene expression. Journal of General Virology 57:1–20
     [Google Scholar]
  13. Ehrlich M., Wang R. Y. H. 1981; 5-methylcytosine in eukaryotic DNA. Science 212:1350–1357
     [Google Scholar]
  14. Gautsch J. W., Wilson M. C. 1983; Delayed de novo methylation in teratocarcinoma suggests additional tissue specific mechanisms for controlling gene expression. Nature; London: 30132–37
     [Google Scholar]
  15. Geder L. 1976; Evidence for early nuclear antigens in cytomegalovirus infected cells. Journal of General Virology 32:315–319
     [Google Scholar]
  16. Geelen J. L. M. C., Boom R., Klaver G. P. M., Minnaar R. P., Feltkamp M. C. W., Van Milligen F. J., Sol C. J. A., Van Der Noordaa J. 1987; Transcriptional activation of the major immediate early transcription unit of human cytomegalovirus by heat-shock, arsenite and protein synthesis inhibitors. Journal of General Virology 68:2925–2931
     [Google Scholar]
  17. Gibson W. 1981; Immediate early proteins of human cytomegalovirus AD169, Davis and Towne differ in electrophoretic mobility. Virology 112:350–354
     [Google Scholar]
  18. Goldstein L. C., Mcdougall J., Hackman R., Meyers J. D., Thomas E. D., Nowinsky R. C. 1982; Monoclonal antibodies to cytomegalovirus: rapid identification of clinical isolates and preliminary use in diagnosis of cytomegalovirus pneumonia. Infection and Immunity 38:273–281
     [Google Scholar]
  19. Gönczöl E., Andrews P. W., Plotkin S. A. 1984; Cytomegalovirus replicates in differentiated but not in undifferentiated human embryonal carcinoma cells. Science 224:159–161
     [Google Scholar]
  20. Gönczöl E., Andrews P. W., Plotkin S. A. 1985; Cytomegalovirus infection of human teratocarcinoma cells in culture. Journal of General Virology 66:509–515
     [Google Scholar]
  21. Gorman C. M., Rigby P. W., Lane D. P. 1985; Negative regulation of viral enhancers in undifferentiated embryonic stem cells. Cell 42:519–526
     [Google Scholar]
  22. Groudine M., Conklin K. F. 1985; Chromatin structure and de novo methylation of sperm DNA: implications for activation of the paternal genome. Science 228:1061–1068
     [Google Scholar]
  23. Hamilton J. D. 1982; Cytomegalovirus and immunity. Monographs in Virology 12:21–24
     [Google Scholar]
  24. Harbers K., Schnieke A., Stuhlmann H., Jähner D., Jaenisch R. 1981; DNA methylation and gene expression: endogenous retroviral genome becomes infectious after molecular cloning. Proceedings of the National Academy of Sciences, U.S.A 78:7609–7613
     [Google Scholar]
  25. Jaenisch R., Jähner D. 1984; Methylation, expression and chromosomal position of genes in mammals. Biochimica et biophysica acta 782:1–9
     [Google Scholar]
  26. Jaenisch R., Schnieke A., Harbers K. 1985; Treatment of mice with 5-azacytidine efficiently activates silent retroviral genomes in different tissues. Proceedings of the National Academy of Sciences, U.S.A 82:1451–1455
     [Google Scholar]
  27. Jahn G. E., Knust E., Schmolla H., Sarre T., Nelson J. A., Mcdougall J. K., Fleckenstein B. 1984; Predominant immediate early transcripts of human cytomegalovirus AD 169. Journal of Virology 49:363–370
     [Google Scholar]
  28. Jähner D., Jaenisch R. 1985a; Retrovirus induced de novo methylation of flanking host sequences correlates with gene inactivity. Nature; London: 315594–597
     [Google Scholar]
  29. Jähner D., Jaenisch R. 1985b; Chromosomal position and specific demethylation in enhancer sequences of germ line-transmitted retroviral genomes during mouse development. Molecular and Cellular Biology 5:2212–2220
     [Google Scholar]
  30. Jordan M. C. 1983; Latent infection and the elusive cytomegalovirus. Reviews of Infectious Diseases 5:205–215
     [Google Scholar]
  31. Kamata T., Tanaka S., Watanabe Y. 1978; Human cytomegalovirus induced chromatin factors responsible for changes in template activity and structure of infected cell chromatin. Virology 90:197–208
     [Google Scholar]
  32. Kamata T., Tanaka S., Watanabe Y. 1979; Characterization of the human cytomegalovirus-induced factor responsible for activation of host cell chromatin template. Virology 97:224–228
     [Google Scholar]
  33. Kierszenbaum A. L., Huang E. S. 1978; Chromatin pattern consisting of repeating bipartite structures in WI-38 cells infected with human cytomegalovirus. Journal of Virology 28:661–664
     [Google Scholar]
  34. Lafemina R., Hayward G. 1986; Constitutive and retinoic acid-inducible expression of cytomegalovirus immediate early genes in human teratocarcinoma cells. Journal of Virology 58:434–440
     [Google Scholar]
  35. Littlefield J. W. 1964; Selection of hybrids from matings of fibroblasts in vitro and their presumed recombinants. Science 145:709–710
     [Google Scholar]
  36. Mcclelland M., Nelson M. 1985; The effect of site specific methylation on restriction endonuclease digestion. Nucleic Acids Research 13:r201–r207
     [Google Scholar]
  37. Mcdonough S. H., Spector D. H. 1983; Transcription in human fibroblasts permissively infected by human cytomegalovirus strain AD169. Virology 125:31–46
     [Google Scholar]
  38. Michelson-Fiske S., Horodniceanu F., Guillon J. C. 1977; Immediate early antigens in human cytomegalovirus infected cells. Nature; London: 270615–617
     [Google Scholar]
  39. Michelson S., Horodniceanu F., Kress M., Tardy-Panit M. 1979; Human cytomegalovirus-induced immediate early antigens: analysis in sodium dodecylsulfate polyacrylamide gel electrophoresis after immunoprecipitation. Journal of Virology 32:259–267
     [Google Scholar]
  40. Nelson J. A., Groudine M. 1986; Transcriptional regulation of the human cytomegalovirus major immediate early gene is associated with induction of DNAse I hypersensitive sites. Molecular and Cellular Biology 6:452–461
     [Google Scholar]
  41. Nelson M., Christ C., Schildkraut J. 1984; Alteration of apparent restriction endonuclease recognition specificities by DNA methylases. Nucleic Acids Research 12:5165–5173
     [Google Scholar]
  42. Niwa O., Yokota Y., Ishida H., Sugahara T. 1983; Independent mechanisms involved in suppression of the Moloney leukemia virus genome during differentiation of murine teratocarcinoma cells. Cell 32:1105–1113
     [Google Scholar]
  43. Razin A., Szyf M. 1984; DNA methylation patterns.Formation and function. Biochimica et biophysica acta 782:331–342
     [Google Scholar]
  44. Reeves R. 1984; Transcriptionally active chromatin. Biochimica et biophysica acta 782:343–393
     [Google Scholar]
  45. Reynolds D. W. 1978; Development of early nuclear antigen in cytomegalovirus infected cells in the presence of RNA and protein synthesis inhibitors. Journal of General Virology 40:475–480
     [Google Scholar]
  46. Riggs A. D., Jones P. 1983; 5-methylcytosine, gene regulation and cancer. Advances in Cancer Research 40:1–30
     [Google Scholar]
  47. Simon D., Stuhlmann H., Jähner D., Wagner H., Werner E., Jaenisch R. 1983; Retrovirus genomes methylated by mammalian but not bacterial methylase are non-infectious. Nature; London: 304275–277
     [Google Scholar]
  48. Smith H. O., Birnstiel M. L. 1976; A simple method for DNA restriction site mapping. Nucleic Acids Research 3:2387–2398
     [Google Scholar]
  49. Spaete R. R., Mocarski E. S. 1985; Regulation of cytomegalovirus gene expression: α and β promoters are trans activated by viral functions in permissive human fibroblasts. Journal of Virology 56:135–143
     [Google Scholar]
  50. Spector D. J., Tevethia M. 1986; Identification of a human cytomegalovirus DNA segment that complements an adenovirus 5. Immediate early mutant. Virology 151:329–338
     [Google Scholar]
  51. Stenberg R. M., Stinski M. F. 1985; Autoregulation of the human cytomegalovirus major immediate early gene. Journal of Virology 56:676–682
     [Google Scholar]
  52. Stenberg R. M., Thomsen D. R., Stinski M. F. 1984; Structural analysis of the major immediate early gene of human cytomegalovirus. Journal of Virology 49:190–199
     [Google Scholar]
  53. Stenberg R. M., Witte P. R., Stinski M. F. 1985; Multiple spliced and unspliced transcripts from human cytomegalovirus immediate-early region 2 and evidence for a common initiation site within immediate-early region 1. Journal of Virology 56:665–675
     [Google Scholar]
  54. Stinski M. F. 1978; Sequence of protein synthesis in cells infected by human cytomegalovirus: early and late virus-induced polypeptides. Journal of Virology 26:686–701
     [Google Scholar]
  55. Stinski M. F., Roehr T. J. 1985; Activation of the major immediate early gene of human cytomegalovirus by exacting elements in the promoter-regulatory sequence and by virus-specific trans-acting components. Journal of Virology 55:431–444
     [Google Scholar]
  56. Stinski M. F., Thomsen D. R., Stenberg R. M., Goldstein L. C. 1983; Organisation and expression of the immediate early genes of human cytomegalovirus. Journal of Virology 46:1–14
     [Google Scholar]
  57. Tanaka S., Otsuka M., Ihara S., Maeda F., Watanabe Y. 1979; Induction of pre-early nuclear antigen(s) in HEL cells infected with human cytomegalovirus. Microbiology and Immunology 23:263–271
     [Google Scholar]
  58. Thomsen D. R., Stenberg R. M., Goins W. F., Stinski M. F. 1984; Promoter-regulatory region of the major immediate early gene of human cytomegalovirus. Proceedings of the National Academy of Sciences, U.S.A 81:659–663
     [Google Scholar]
  59. Topp W. C. 1981; Normal rat cell lines deficient in nuclear thymidine kinase. Virology 113:408–411
     [Google Scholar]
  60. Vardimon L., Rich A. 1984; In Z-DNA the sequence GCGC is neither methylated by HhaI methyltransferase nor cleaved by HhaI restriction endonuclease. Proceedings of the National Academy of Sciences, U.S.A 81:3268–3272
     [Google Scholar]
  61. Wathen M. W., Stinski M. F. 1982; Temporal pattern of human cytomegalovirus transcription: mapping the viral RNA synthesized at immediate early, early and late times after infection. Journal of Virology 41:462–477
     [Google Scholar]
  62. Wathen M. W., Thomsen D. R., Stinski M. F. 1981; Temporal regulation of human cytomegalovirus transcription at immediate early and early times after infection. Journal of Virology 38:446–459
     [Google Scholar]
  63. Weintraub H. 1985; Assembly and propagation of repressed and derepressed chromosomal states. Cell 42:705–711
     [Google Scholar]
  64. Wilkinson G. W. G., Akrigg A., Greenaway P. 1984; Transcription of the immediate early genes of human cytomegalovirus strain AD169. Virus Research 1:101–116
     [Google Scholar]
  65. Wolf S. F., Migeon B. R. 1985; Clusters of CpGdinucleotides implicated by nuclease hypersensitivity as control elements of housekeeping genes. Nature; London: 314467–469
     [Google Scholar]
  66. Youssouffian H., Hammer S. C., Hirsch M. S., Mulder C. 1982; Methylation of the viral genome in an in vitro model of herpes simplex virus latency. Proceedings of the National Academy of Sciences, U.S.A 79:2207–2210
     [Google Scholar]
  67. Zacharias W., Larson J. E., Kilpatrick M. W., Welles R. D. 1984; HhaI methylase and restriction endonuclease as probes for B to Z DNA conformational changes in d(GCGC) sequences. Nucleic Acids Research 12:7677–7692
     [Google Scholar]
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Resistance to Methylation de novo of the Human Cytomegalovirus Immediate Early Enhancer in a Model for Virus Latency and Reactivation in vitro
J. Gen. Virol. 68, 2839 (1987); https://doi.org/10.1099/0022-1317-68-11-2839
/content/journal/jgv/10.1099/0022-1317-68-11-2839
/content/journal/jgv/10.1099/0022-1317-68-11-2839
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