1887

Abstract

We have shown previously that the EBNA 1 and latent membrane protein encoding regions of the Epstein—Barr virus (EBV) genome are highly methylated at CCGG sequences in the Burkitt's lymphoma (BL)-derived cell line Rael, but are unmethylated in a lymphoblastoid cell line (LCL) harbouring the same virus. To examine whether this is a regular phenomenon, we compared the methylation patterns of selected regions (HI C, W, H, M, E, K and N fragments) of EBV DNA in representative EBV-carrying cell types of normal and neoplastic origin. Analysis of II and I cleavage patterns showed that all probed regions were highly methylated in all six BL biopsy samples, but hypomethylated in the four LCLs immortalized by the virus. EBV DNA was also highly methylated in the nude mouse-passaged C15 nasopharyngeal carcinoma strain and partially methylated in the C18 strain. Eight BL lines propagated , ranging from a typical BL group I to a more LCL-like group III phenotype, showed heterogeneous levels of methylation. Rael, the only stable group I cell line, carried highly methylated viral genomes. The other cell lines, which have drifted to an LCL-like blastic phenotype to various degrees, showed more moderate or low viral DNA methylation. Two sublines of the BL cell line Jijoye, which could be classified as groups II and III, respectively, showed a corresponding difference in EBV DNA methylation. To assess the possible influence of hypomethylated linear EBV DNA molecules produced in lytically infected cells, the virus producer P3HR-1 and Jijoye M13 lines were compared for DNA methylation before and after treatment with phosphonoformic acid (PFA), an inhibitor of the viral DNA polymerase. PFA treatment resulted in a shift towards a more methylated pattern in both regions (HI W and E) assayed, but had no effect on virus non-producer lines (Rael, CB-M1-Ral-STO and Jijoye p79).

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-7-1591
1991-07-01
2022-08-16
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/7/JV0720071591.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-7-1591&mimeType=html&fmt=ahah

References

  1. Allday M. J., Kundu D., Finerty S., Griffin B. E. 1990; CpG methylation of viral DNA in EBV-associated tumours. International Journal of Cancer 45:1125–1130
    [Google Scholar]
  2. Busson P., Ganem G., Flores P., Mugneret F., Clausse B., Caillou B., Braham K., Wakasugi H., Pipinski M., Tursz T. 1988; Establishment and characterization of three transplantable EBV-containing nasopharyngeal carcinomas. International Journal of Cancer 42:599–606
    [Google Scholar]
  3. Cedar H. 1988; DNA methylation and gene activity. Cell 53:3–4
    [Google Scholar]
  4. Desrosiers R. C., Mulder C., Fleckenstein B. 1979; Methylation of herpesvirus saimiri DNA in lymphoid tumour cell lines. Proceedings of the National Academy of Sciences, U.S.A 76:3839–3843
    [Google Scholar]
  5. Doerfler W. 1981; DNA methylation - a regulatory signal in eukaryotic gene expression. Journal of General Virology 57:1–20
    [Google Scholar]
  6. Doerfler W. 1983; DNA methylation and gene activity. Annual Review of Biochemistry 52:93–124
    [Google Scholar]
  7. Ehlin-Henriksson B., Klein G. 1984; Distinction between Burkitt lymphoma subgroups by monoclonal antibodies: relationships between antigen expression and type of chromosomal translocation. International Journal of Cancer 33:459–463
    [Google Scholar]
  8. Ernberg I., Kallin B., Dillner J., Falk K., Ehlin-Henriksson B., Hammarskjöld M. L., Klein G. 1986; Lymphoblastoid cell lines and Burkitt-lymphoma-derived cell lines differ in the expression of a second Epstein–Barr virus-encoded nuclear antigen. International Journal of Cancer 38:729–737
    [Google Scholar]
  9. Ernberg I., Falk K., Minarovits J., Busson P., Tursz T., Masucci M. G., Klein G. 1989; The role of methylation in the phenotype-dependent modulation of Epstein–Barr nuclear antigen 2 and latent membrane protein genes in cells latently infected with Epstein–Barr virus. Journal of General Virology 70:2989–3002
    [Google Scholar]
  10. Fåhraeus R., Hu L. F., Ernberg I., Finke J., Rowe M., Klein G., Falk K., Nilsson E., Yadav M., Busson P., Tursz T., Kallin B. 1988; Expression of Epstein–Barr virus proteins in nasopharyngeal carcinoma. International Journal of Cancer 42:329–338
    [Google Scholar]
  11. Fåhraeus R., Jansson A., Ricksten A., Sjöblom A., Rymo L. 1990; Epstein–Barr virus-encoded nuclear antigen 2 activates the viral latent membrane protein promoter by modulating the activity of a negative regulatory element. Proceedings of the National Academy of Sciences, U.S.A 87:7390–7394
    [Google Scholar]
  12. Gompels U. A., Craxton M. A., Honess R. W. 1988; Conservation of gene organization in the lymphotropic herpesviruses herpesvirus saimiri and Epstein–Barr virus. Journal of Virology 62:757–767
    [Google Scholar]
  13. Goodman S. R., Prezyna G., Clough-Benz W. 1978; Two Epstein–Barr virus-associated DNA polymerase activities. Journal of Biological Chemistry 253:8617–8628
    [Google Scholar]
  14. Gunven P., Klein G., Clifford P., Singh S. 1974; Epstein–Barr virus-associated membrane-reactive antibodies during long-term survival after Burkitt’s lymphoma. Proceedings of the National Academy of Sciences, U.S.A 71:1422–1426
    [Google Scholar]
  15. Hammerschmidt W., Sugden B. 1988; Identification and characterization of oriLyt, a lytic origin of DNA replication of Epstein–Barr virus. Cell 56:427–433
    [Google Scholar]
  16. 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 DN As may be diagnostic of differences in the states of their latent genomes. Journal of General Virology 70:837–855
    [Google Scholar]
  17. Hu L. F., Minarovits J., Cao S. L., Contreras-Salazar B., Rymo L., Falk K., Klein G., Ernberg I. 1991; The variable expression of LMP (the latent membrane protein) in nasopharyngeal carcinoma (NPC) can be related to the methylation status of the Epstein–Barr virus BNLF 1 5′-flanking region. Journal of Virology (in press)
    [Google Scholar]
  18. Jones M. D., Foster L., Sheedy T., Griffin B. E. 1984; The EB virus genome in Daudi Burkitt’s lymphoma cells has a deletion similar to that observed in a non-transforming strain (P3HR-1) of the virus. EMBO Journal 3:813–821
    [Google Scholar]
  19. Kallin B., Dillner J., Ernberg I., Ehlin-Henriksson B., Rosen A., Henle W., Henle G., Klein G. 1986; Four virally determined nuclear antigens are expressed in Epstein–Barr virus-transformed cells. Proceedings of the National Academy of Sciences, U.S.A 83:1499–1503
    [Google Scholar]
  20. Klein G. 1989; Viral latency and transformation: the strategy of Epstein–Barr virus. Cell 58:5–8
    [Google Scholar]
  21. Lewin N., Minarovits J., Weber G., Ehlin-Henriksson B., Wen T., Mellstedt H., Klein G., Klein E. 1991; Clonality and methylation status of the Esptein–Barr virus (EBV) genomes in in vivo infected EBV-carrying chronic lymphocytic leukemia (CLL) cell lines. International Journal of Cancer (in press)
    [Google Scholar]
  22. Maniatis T., Fritsch E. F., Sambrook J. 1982; Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  23. 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. Journal of Virology 63:3135–3141
    [Google Scholar]
  24. Öberg B. 1983; Antiviral effects of phosphonoformate (PFA, Foscamet sodium). Pharmacological Therapeutics 19:387–415
    [Google Scholar]
  25. Olweny Cm. L. M., Atine I., Kaddu-Mukasa A., Owor R., Andersson-Anvret M., Klein G., Henle W., de The G. 1977; Epstein–Barr virus genome studies in Burkitt’s and non-Burkitt’s lymphomas in Uganda. Journal of the National Cancer Institute 58:1191–1196
    [Google Scholar]
  26. Perlmann C., Saemundsen A. K., Klein G. 1982; A fraction of Epstein–Barr virus DNA is methylated in and around the Eco RI-J fragment. Virology 123:217–221
    [Google Scholar]
  27. Ricksten A., Olsson A., Andersson T., Rymo L. 1988; The 5′ flanking region of the gene for the Epstein–Barr virus-encoded nuclear antigen 2 contains a cell type-specific cis-acting regulatory element that activates transcription in transfected B-cells. Nucleic Acids Research 16:8391–8409
    [Google Scholar]
  28. Rowe D., Heston L., Metlay J., Miller G. 1985; Identification and expression of a nuclear antigen from the genomic region of the Jijoye strain of Epstein–Barr virus that is missing in its nonimmortalizing deletion mutant, P3HR-1. Proceedings of the National Academy of Sciences, U.S.A 82:7429–7433
    [Google Scholar]
  29. Rowe M., Rowe D. T., Gregory C. D., Young L. S., Farrell P. J., Rupani H., Rickinson A. D. 1987; Differences in B cell growth phenotype reflect novel patterns of Epstein–Barr virus latent gene expression in Burkitt’s lymphoma cells. EMBO Journal 6:2743–2751
    [Google Scholar]
  30. Shaw J. E. 1985; The circular intracellular form of Epstein–Barr virus DNA is amplified by the virus-associated DNA polymerase. Journal of Virology 53:1012–1015
    [Google Scholar]
  31. Summers W. C., Klein G. 1976; Inhibition of Epstein–Barr virus DNA synthesis and late gene expression by phosphonoacetic acid. Journal of Virology 18:151–155
    [Google Scholar]
  32. Szyf M., Eliasson L., Mann V., Klein G., Razin A. 1985; Cellular and viral DNA hypomethylation associated with induction of Epstein–Barr virus lytic cycle. Proceedings of the National Academy of Sciences, U.S.A 82:8090–8094
    [Google Scholar]
  33. Waalwijk C., Flavell R.A. 1978; Msp I, an isoschizomer of Hpa II which cleaves both unmethylated and methylated Hpa II sites. Nucleic Acids Research 5:3231–3236
    [Google Scholar]
  34. Yates J., Warren N., Reisman D., Sugden B. 1984; A cis-acting element from the Epstein–Barr viral genome that permits stable replication of recombinant plasmids in latently infected cells. Proceedings of the National Academy of Sciences, U.S.A 81:3806–3810
    [Google Scholar]
  35. Young L. S., Yao Q. Y., Rooney C. M., Sculley T. B., Moss D. J., Rupani H., Laux G., Bornkamm G. W., Rickinson A. B. 1987; New type B isolates of Epstein–Barr virus from Burkitt’s lymphoma and from normal individuals in endemic areas. Journal of General Virology 68:2853–2862
    [Google Scholar]
  36. Yousuffian H., Hammer S. M., 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]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-7-1591
Loading
/content/journal/jgv/10.1099/0022-1317-72-7-1591
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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