1887

Abstract

We have used a spleen explant model to investigate mechanisms of murine cytomegalovirus latency and reactivation. Induction of immediate-early () gene expression occurs in explants after approximately 9 days in culture and virus reactivation follows induction of gene expression with kinetics similar to that of productive infection . This occurs independently of TNF receptor signalling. Treatment with the DNA methylation inhibitor 5-aza-2′-deoxycytidine and the histone deacetylase inhibitor trichostatin A results in more rapid induction of gene expression and reactivation of virus. Despite these results, which suggest a role for DNA methylation in maintenance of viral latency, we find that the major immediate-early promoter/enhancer is not methylated in latently infected mice. Our results support the hypothesis that latency is maintained by epigenetic control of gene expression, and that induction of gene expression leads to reactivation of virus, but suggest that these are not controlled by DNA methylation.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.82696-0
2007-04-01
2019-11-13
Loading full text...

Full text loading...

/deliver/fulltext/jgv/88/4/1097.html?itemId=/content/journal/jgv/10.1099/vir.0.82696-0&mimeType=html&fmt=ahah

References

  1. Bubic, I., Wagner, M., Krmpotic, A., Saulig, T., Kim, S., Yokoyama, W. M., Jonjic, S. & Koszinowski, U. H. ( 2004; ). Gain of virulence caused by loss of a gene in murine cytomegalovirus. J Virol 78, 7536–7544.[CrossRef]
    [Google Scholar]
  2. Chen, J., Ueda, K., Sakakibara, S., Okuno, T., Parravicini, C., Corbellino, M. & Yamanishi, K. ( 2001; ). Activation of latent Kaposi's sarcoma-associated herpesvirus by demethylation of the promoter of the lytic transactivator. Proc Natl Acad Sci U S A 98, 4119–4124.[CrossRef]
    [Google Scholar]
  3. Clark, S. J., Harrison, J., Paul, C. L. & Frommer, M. ( 1994; ). High sensitivity mapping of methylated cytosines. Nucleic Acids Res 22, 2990–2997.[CrossRef]
    [Google Scholar]
  4. Dorsch-Hasler, K., Keil, G. M., Weber, F., Jasin, M., Schaffner, W. & Koszinowski, U. H. ( 1985; ). A long and complex enhancer activates transcription of the gene coding for the highly abundant immediate-early mRNA in murine cytomegalovirus. Proc Natl Acad Sci U S A 82, 8325–8329.[CrossRef]
    [Google Scholar]
  5. El Kharroubi, A., Piras, G. & Stewart, C. L. ( 2001; ). DNA demethylation reactivates a subset of imprinted genes in uniparental mouse embryonic fibroblasts. J Biol Chem 276, 8674–8680.[CrossRef]
    [Google Scholar]
  6. Grassi, G., Maccaroni, P., Meyer, R., Kaiser, H., D'Ambrosio, E., Pascale, E., Grassi, M., Kuhn, A., Di Nardo, P. & other authors ( 2003; ). Inhibitors of DNA methylation and histone deacetylation activate cytomegalovirus promoter-controlled reporter gene expression in human glioblastoma cell line U87. Carcinogenesis 24, 1625–1635.[CrossRef]
    [Google Scholar]
  7. Herman, J. G. & Baylin, S. B. ( 2003; ). Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med 349, 2042–2054.[CrossRef]
    [Google Scholar]
  8. Herman, J. G., Graff, J. R., Myohanen, S., Nelkin, B. D. & Baylin, S. B. ( 1996; ). Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci U S A 93, 9821–9826.[CrossRef]
    [Google Scholar]
  9. Honess, R. W., Gompels, U. A., Barrell, B. G., Craxton, M., Cameron, K. R., Staden, R., Chang, Y. N. & Hayward, G. S. ( 1989; ). Deviations from expected frequencies of CpG dinucleotides in herpesvirus DNAs may be diagnostic of differences in the states of their latent genomes. J Gen Virol 70, 837–855.[CrossRef]
    [Google Scholar]
  10. Hummel, M., Zhang, Z., Yan, S., DePlaen, I., Golia, P., Varghese, T., Thomas, G. & Abecassis, M. I. ( 2001; ). Allogeneic transplantation induces expression of cytomegalovirus immediate-early genes in vivo: a model for reactivation from latency. J Virol 75, 4814–4822.[CrossRef]
    [Google Scholar]
  11. Jones, P. A. ( 1999; ). The DNA methylation paradox. Trends Genet 15, 34–37.[CrossRef]
    [Google Scholar]
  12. Jordan, M. C. & Mar, V. L. ( 1982; ). Spontaneous activation of latent cytomegalovirus from murine spleen explants. Role of lymphocytes and macrophages in release and replication of virus. J Clin Invest 70, 762–768.[CrossRef]
    [Google Scholar]
  13. Koffron, A. J., Hummel, M., Patterson, B. K., Yan, S., Kaufman, D. B., Fryer, J. P., Stuart, F. P. & Abecassis, M. I. ( 1998; ). Cellular localization of latent murine cytomegalovirus. J Virol 72, 95–103.
    [Google Scholar]
  14. Kurz, S. K., Rapp, M., Steffens, H. P., Grzimek, N. K., Schmalz, S. & Reddehase, M. J. ( 1999; ). Focal transcriptional activity of murine cytomegalovirus during latency in the lungs. J Virol 73, 482–494.
    [Google Scholar]
  15. Magdinier, F. & Wolffe, A. P. ( 2001; ). Selective association of the methyl-CpG binding protein MBD2 with the silent p14/p16 locus in human neoplasia. Proc Natl Acad Sci U S A 98, 4990–4995.[CrossRef]
    [Google Scholar]
  16. Masucci, M. G., Contreras-Salazar, B., Ragnar, E., Falk, K., Minarovits, J., Ernberg, I. & Klein, G. ( 1989; ). 5-Azacytidine up regulates the expression of Epstein-Barr virus nuclear antigen 2 (EBNA-2) through EBNA-6 and latent membrane protein in the Burkitt's lymphoma line rael. J Virol 63, 3135–3141.
    [Google Scholar]
  17. Meier, J. L. ( 2001; ). Reactivation of the human cytomegalovirus major immediate-early regulatory region and viral replication in embryonal NTera2 cells: role of trichostatin A, retinoic acid, and deletion of the 21-base-pair repeats and modulator. J Virol 75, 1581–1593.[CrossRef]
    [Google Scholar]
  18. Mocarski, E. S. & Courcell, C. T. ( 2001; ). Cytomegaloviruses and their replication. In Fields Virology, 4th edn, pp. 2629–2673. Edited by D. M. Knipe & P. M. Howley. Philadelphia, PA: Lippincott Williams & Wilkins.
  19. Nguyen, C. T., Weisenberger, D. J., Velicescu, M., Gonzales, F. A., Lin, J. C., Liang, G. & Jones, P. A. ( 2002; ). Histone H3-lysine 9 methylation is associated with aberrant gene silencing in cancer cells and is rapidly reversed by 5-aza-2′-deoxycytidine. Cancer Res 62, 6456–6461.
    [Google Scholar]
  20. Raizis, A. M., Schmitt, F. & Jost, J. P. ( 1995; ). A bisulfite method of 5-methylcytosine mapping that minimizes template degradation. Anal Biochem 226, 161–166.[CrossRef]
    [Google Scholar]
  21. Rawlinson, W. D., Farrell, H. E. & Barrell, B. G. ( 1996; ). Analysis of the complete DNA sequence of murine cytomegalovirus. J Virol 70, 8833–8849.
    [Google Scholar]
  22. Reeves, M. B., MacAry, P. A., Lehner, P. J., Sissons, J. G. & Sinclair, J. H. ( 2005; ). Latency, chromatin remodeling, and reactivation of human cytomegalovirus in the dendritic cells of healthy carriers. Proc Natl Acad Sci U S A 102, 4140–4145.[CrossRef]
    [Google Scholar]
  23. Rodic, N., Oka, M., Hamazaki, T., Murawski, M. R., Jorgensen, M., Maatouk, D. M., Resnick, J. L., Li, E. & Terada, N. ( 2005; ). DNA methylation is required for silencing of Ant4, an adenine nucleotide translocase selectively expressed in mouse embryonic stem cells and germ cells. Stem Cells 23, 1314–1323.[CrossRef]
    [Google Scholar]
  24. Takebayashi, S., Nakao, M., Fujita, N., Sado, T., Tanaka, M., Taguchi, H. & Okumura, K. ( 2001; ). 5-Aza-2′-deoxycytidine induces histone hyperacetylation of mouse centromeric heterochromatin by a mechanism independent of DNA demethylation. Biochem Biophys Res Commun 288, 921–926.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.82696-0
Loading
/content/journal/jgv/10.1099/vir.0.82696-0
Loading

Data & Media loading...

Most Cited This Month

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