1887

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Volume 69, Issue 6

Induction of Gene Expression under Human Cytomegalovirus Immediate Early Enhancer-Promoter Control by Inhibition of Protein Synthesis Is Cell Cycle-dependent Free

Abstract

Summary

In this paper we describe stably transfected rat cell lines which harbour either the human cytomegalovirus (HCMV) immediate early (IE) gene encoding the 72K IE nuclear antigen (IEA) or the bacterial chloramphenicol acetyltransferase (CAT) gene both under transcriptional control of the HCMV IE enhancer-promoter (-484 to -19 relative to the IE cap site, +1). In these cell lines IE gene or CAT gene expression is repressed but can be induced by heat-shock, by sodium arsenite and by inhibitors of protein synthesis such as cycloheximide (CH). In addition, we present evidence suggesting that CH-mediated activation is cell cycle-dependent. Thus CH-mediated induction of the 72K IEA as well as CAT gene expression was impaired and accumulation of mRNAs did not occur when cellular DNA synthesis was inhibited. Activation of IE genes by CH occurred almost exclusively in those cells which were in S-phase. In contrast, activation of gene expression by sodium arsenite occurred independently of cellular DNA synthesis and was not restricted to cells in S-phase. The data are consistent with, but not proof of, the hypothesis that the activation of IE transcription, brought about by inhibition of protein synthesis, resulted from a disturbed chromatin conformation due to DNA synthesis continuing in the absence of a supply of chromatin-organizing proteins. The possible relevance of these observations with regard to HCMV latency and reactivation is discussed.

  • Received:
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Keyword(s): CMV , human , IE enhancer and S-phase
© Society for General Microbiology 1988
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1988-06-01
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. ANNUNZIATO A. T., SEALE R. L. 1983; Chromatin replication, reconstitution and assembly. Molecular and Cellular Biochemistry 55:99–112
     [Google Scholar]
  3. ANNUNZIATO A. T., SEALE R. 1984; Presence of nucleosomes within irregularly cleaved fragments of newly replicated chromatin. Nucleic Acids Research 12:6179–6196
     [Google Scholar]
  4. ASHBURNER M., BONNER J. J. 1979; The induction of gene activity in Drosophila by heat-shock. Cell 17:241–254
     [Google Scholar]
  5. BIRD A. P. 1986; CpG-rich islands and the function of DNA methylation. Nature, London 321:209–213
     [Google Scholar]
  6. BOOM R. 1987 Repression and activation of the human cytomegalovirus major immediate early transcription unit Ph.D. Thesis, University of Amsterdam
    [Google Scholar]
  7. BOOM R., GEELEN J. L., SOL C. J. A., 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. BOOM R., GEELEN J. L, SOL C. J., MINNAAR R. P., VAN DER NOORDAA J. 1987; Resistance to methylation de novo of the human cytomegalovirus immediate early enhancer in a model for virus latency and reactivation in vitro. Journal of General Virology 68:2839–2852
     [Google Scholar]
  9. 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]
  10. BROWN D. P. 1984; The role of stable complexes that repress and activate eukaryotic genes. Cell 37:359–365
     [Google Scholar]
  11. BURDON R. 1986; Heat-shock and the heat-shock proteins. Biochemical Journal 240:313–324
     [Google Scholar]
  12. COLBÈRE-GARAPIN F., CHOUSTERMAN S., HORODNICEANU F., KOURILSKY P., GARAPIN A. 1979; Cloning of the active thymidine kinase gene of herpes simplex virus type 1 in E. coli Kl2. Proceedings of the National Academy of Sciences, U.S.A 76:3755–3759
     [Google Scholar]
  13. COZZARELLI N. R. 1977; The mechanism of action of inhibitors of DNA synthesis. Annual Review of Biochemistry 46:641–668
     [Google Scholar]
  14. DANI C, BLANCHARD J. M., PIECHACZYK M., EL SABOUTY S., MARTY L., JEANTEUR P. 1984; Extreme instability of myc mRNA in normal and transformed human cells. Proceedings of the National Academy of Sciences, U.S.A 81:7046–7050
     [Google Scholar]
  15. ELDER P. K., SCHMIDT L. J., ONO T., GETZ M. J. 1984; Specific stimulation of actin gene transcription by epidermal growth factor and cycloheximide. Proceedings of the National Academy of Sciences, U.S.A 81:7476–7480
     [Google Scholar]
  16. GEELEN I. L. M. C, BOOM R., KLAVER G. P. M., MINNAAR R. P., FELTKAMP M. C. W., VAN MILLIGEN F. J., SOL C. I. 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. GLEAVES C A., SMITH T. F., SHUSTER E. A., PEARSON G. R. 1985; Comparison of standard tube and shell vial culture techniques for the detection of cytomegalovirus in clinical specimens. Journal of Clinical Microbiology 21:217–221
     [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, PADMANABHAN R., HOWARD B. H. 1983; High efficiency DNA-mediated transformation of primate cells. Science 221:551–553
     [Google Scholar]
  22. GRAHAM R. C. 1965; Cytochemical demonstration of peroxidase activity with 3-amino-9-ethylcarbazole. Journal of Histochemistry and Cytochemistry 13:150–152
     [Google Scholar]
  23. GREENBERG M. E., HERMANOWSKY A. L., ZIFF E. B. 1986; Effect of protein synthesis inhibitors on growth factor activation of c-fos, c-myc and actin gene transcription. Molecular and Cellular Biology 6:1050–1057
     [Google Scholar]
  24. HENNIGHAUSEN L., FLECKENSTEIN B. 1986; Nuclear factor 1 interacts with five DNA elements in the promoter region of the human cytomegalovirus major immediate early gene. EMBO Journal 5:1367–1371
     [Google Scholar]
  25. 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 AD169. Journal of Virology 49:363–370
     [Google Scholar]
  26. JORDAN M. C. 1983; Latent infection and the elusive cytomegalovirus. Reviews of Infectious Diseases 5:205–215
     [Google Scholar]
  27. 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]
  28. LINIAL M., GUNDERSON N., GROUDINE M. 1985; Enhanced transcription of c-myc in bursal lymphoma cells requires continuous protein synthesis. Science 230:1126–1132
     [Google Scholar]
  29. LITTLEFIELD J. W. 1964; Selection of hybrids from matings of fibroblasts in vitro and their presumed recombinants. Science 145:709–710
     [Google Scholar]
  30. MCDONOUGH S. H., SPECTOR D. H. 1983; Transcription in human fibroblasts permissively infected by human cytomegalovirus strain AD169. Virology 125:31–46
     [Google Scholar]
  31. MAXAM A. M., GILBERT W. 1980; Sequencing end-labelled DNA with base-specific chemical cleavages. Methods in Enzymology 65:499–560
     [Google Scholar]
  32. MUNRO S., PELHAM H. 1985; What turns on heat-shock genes?. Nature, London 317:477–478
     [Google Scholar]
  33. 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]
  34. PELHAM H. 1985; Activation of heat-shock genes in eukaryotes. Trends in Genetics 1:31–35
     [Google Scholar]
  35. PIGNATTI P. F., CASSAI E., MENEGUZZI G., CHENCINER N., MILANESI G. 1979; Herpes simplex virus DNA isolation from infected cells with a novel procedure. Virology 93:260–264
     [Google Scholar]
  36. REEVES R. 1984; Transcriptionally active chromatin. Biochimica et biophysica acta 782:343–393
     [Google Scholar]
  37. RILEY D., WEINTRAUB H. 1979; Conservative segregation of parental histones during replication in the presence of cycloheximide. Proceedings of the National Academy of Sciences, U.S.A 76:328–332
     [Google Scholar]
  38. RINGOLD G. M., DIECKMANN B., VANNICE J. L., TRAHEY M., MCCORMICK F. 1984; Inhibition of protein synthesis stimulates the transcription of human β-interferon genes in Chinese hamster ovary cells. Proceedings of the National Academy of Sciences, U.S.A 81:3964–3968
     [Google Scholar]
  39. SCHÜMPERLI D. 1986; Cell-cycle regulation of histone gene expression. Cell 45:471–472
     [Google Scholar]
  40. SEALE R. L., SIMPSON R. T. 1975; Effects of cycloheximide on chromatin biosynthesis. Journal of Molecular Biology 94:479–501
     [Google Scholar]
  41. SHUSTER E. A., BENEKE I. S., TEGTMEYER G. E., PEARSON G. R., CLEAVES C. A., WOLD A. D., SMITH T. F. 1985; Monoclonal antibody for rapid laboratory detection of cytomegalovirus infections: characterization and diagnostic application. Mayo Clinic Proceedings 60:577–585
     [Google Scholar]
  42. SIVE H. L., HEINTZ N., ROEDER R. G. 1984; Regulation of human histone gene expression during the HeLa cell cycle requires protein synthesis. Molecular and Cellular Biology 4:2723–2734
     [Google Scholar]
  43. SPAETE R. R., MOCARSKI E. S. 1985; Regulation of cytomegalovirus gene expression: a and β promoters are transactivated by viral functions in permissive human fibroblasts. Journal of Virology 56:135–143
     [Google Scholar]
  44. STENBERG R. M., STINSKI M. F. 1985; Autoregulation of the human cytomegalovirus major immediate early gene. Journal of Virology 56:676–683
     [Google Scholar]
  45. 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]
  46. 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]
  47. STIMAC E., GROPPI V. E. JR, COFFINO P. 1984; Inhibition of protein synthesis stabilizes histone mRNA. Molecular and Cellular Biology 4:2082–2090
     [Google Scholar]
  48. 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 transacting components. Journal of Virology 55:431–441
     [Google Scholar]
  49. 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]
  50. 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]
  51. TIMSON J. 1975; Hydroxyurea. Mutation Research 32:115–132
     [Google Scholar]
  52. TOPP W. C. 1981; Normal rat cell lines deficient in nuclear thymidine kinase. Virology 113:408–411
     [Google Scholar]
  53. WEINTRAUB H. 1985; Assembly and propagation of repressed and derepressed chromosomal states. Cell 42:705–711
     [Google Scholar]
  54. 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]
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Induction of Gene Expression under Human Cytomegalovirus Immediate Early Enhancer-Promoter Control by Inhibition of Protein Synthesis Is Cell Cycle-dependent
J. Gen. Virol. 69, 1179 (1988); https://doi.org/10.1099/0022-1317-69-6-1179
/content/journal/jgv/10.1099/0022-1317-69-6-1179
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