1887

Abstract

Epstein–Barr virus (EBV) nuclear antigen-1 (EBNA-1), which binds to both the EBV origin of replication () and metaphase chromosomes, is essential for the replication/retention and segregation/partition of -containing plasmids. Here the chromosomal localization of EBNA-1 fused to green fluorescent protein (GFP–EBNA-1) is examined by confocal microscopy combined with a ‘premature chromosome condensation’ (PCC) procedure. Analyses show that GFP–EBNA-1 expressed in living cells that lack plasmids is associated with cellular chromatin that has been condensed rapidly by the PCC procedure into identifiable forms that are unique to each phase of interphase as well as metaphase chromosomes. Studies of cellular chromosomal DNAs labelled with BrdU or Cy3-dUTP indicate that GFP–EBNA-1 colocalizes highly with the labelled, newly replicated regions of interphase chromatin in cells. These results suggest that EBNA-1 is associated not only with cellular metaphase chromosomes but also with condensing chromatin/chromosomes and probably with interphase chromatin, especially with its newly replicated regions.

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2002-10-01
2020-08-03
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References

  1. Aiyar A., Tyree C., Sugden B.. 1998; The plasmid replicon of EBV consists of multiple cis -acting elements that facilitate DNA synthesis by the cell and a viral maintenance element. EMBO Journal17:6394–6403
    [Google Scholar]
  2. Alsbeih G., Raaphorst G. P.. 1999; Differential induction of premature chromosome condensation by calyculin A in human fibroblast and tumor cell lines. Anticancer Research19:903–908
    [Google Scholar]
  3. Ballestas M. E., Chatis P. A., Kaye K. M.. 1999; Efficient persistence of extrachromosomal KSHV DNA mediated by latency-associated nuclear antigen. Science284:641–644
    [Google Scholar]
  4. Carayon P., Bord A.. 1992; Identification of DNA-replicating lymphocyte subsets using a new method to label the bromo-deoxyuridine incorporated into the DNA. Journal of Immunological Methods147:225–230
    [Google Scholar]
  5. Coco-Martin J. M., Begg A. C.. 1997; Detection of radiation-induced chromosome aberrations using fluorescence in situ hybridization in drug-induced premature chromosome condensations of tumour cell lines with different radiosensitivities. International Journal of Radiation Biology71:265–273
    [Google Scholar]
  6. Cook P. R.. 2001; Principles of Nuclear Structure and Function New York: Wiley-Liss;
    [Google Scholar]
  7. Fujita T., Ikeda M., Kusano S., Yamazaki M., Ito S., Obayashi M., Yanagi K.. 2001; Amino acid substitution analyses of the DNA contact region, two amphipathic alpha-helices and a recognition-helix-like helix outside the dimeric beta-barrel of Epstein–Barr virus nuclear antigen 1. Intervirology44:271–282
    [Google Scholar]
  8. Gotoh E., Asakawa Y., Kosaka H.. 1995; Inhibition of protein serine/threonine phosphatases directly induces premature chromosome condensation in mammalian somatic cells. Biomedical Research16:63–68
    [Google Scholar]
  9. Grogan E. A., Summers W. P., Dowling S., Shedd D., Gradoville L., Miller G.. 1983; Two Epstein–Barr viral nuclear neoantigens distinguished by gene transfer, serology, and chromosome binding. Proceedings of the National Academy of Sciences, USA80:7650–7653
    [Google Scholar]
  10. Guo X. W., Th′ng J. P., Swank R. A., Anderson H. J., Tudan C., Bradbury E. M., Roberge M.. 1995; Chromosome condensation induced by fostriecin does not require p34cdc2 kinase activity and histone H1 hyperphosphorylation, but is associated with enhanced histone H2A and H3 phosphorylation. EMBO Journal14:976–985
    [Google Scholar]
  11. Hung S. C., Kang M. S., Kieff E.. 2001; Maintenance of Epstein–Barr virus (EBV) oriP-based episomes requires EBV-encoded nuclear antigen-1 chromosome-binding domains, which can be replaced by high-mobility group-I or histone H1. Proceedings of the National Academy of Sciences, USA98:1865–1870
    [Google Scholar]
  12. Ito S., Ikeda M., Kato N., Matsumoto A., Ishikawa Y., Kumakubo S., Yanagi K.. 2000; Epstein–Barr virus nuclear antigen-1 binds to nuclear transporter karyopherin alpha1/NPI-1 in addition to karyopherin alpha2/Rch1. Virology266:110–119
    [Google Scholar]
  13. Johnson R. T., Rao P. N.. 1970; Mammalian cell fusion: induction of premature chromosome condensation in interphase nuclei. Nature226:717–722
    [Google Scholar]
  14. Kanda T., Otter M., Wahl G. M.. 2001; Coupling of mitotic chromosome tethering and replication competence in Epstein–Barr virus-based plasmids. Molecular and Cellular Biology21:3576–3588
    [Google Scholar]
  15. Kieff E., Rickinson A. B.. 2001; Epstein–Barr virus and its replication. In Fields Virology pp2511–2573 Edited by Knipe D. M., Howley P. M.. Philadelphia: Lippincott Williams & Wilkins;
    [Google Scholar]
  16. Kube D., Vockerodt M., Weber O., Hell K., Wolf J., Haier B., Grasser F. A., Muller-Lantzsch N., Kieff E., Diehl V., Tesch H.. 1999; Expression of Epstein–Barr virus nuclear antigen 1 is associated with enhanced expression of CD25 in the Hodgkin cell line L428. Journal of Virology73:1630–1636
    [Google Scholar]
  17. Kusano S., Tamada K., Senpuku H., Harada S., Ito S., Yanagi K.. 2001; Epstein–Barr virus nuclear antigen-1-dependent and -independent oriP -binding cellular proteins. Intervirology44:283–290
    [Google Scholar]
  18. Mackey D., Sugden B.. 1999; The linking regions of EBNA1 are essential for its support of replication and transcription. Molecular and Cellular Biology19:3349–3359
    [Google Scholar]
  19. McNeil P. L., Warder E.. 1987; Glass beads load macromolecules into living cells. Journal of Cell Science88:669–678
    [Google Scholar]
  20. Manders E. M., Kimura H., Cook P. R.. 1999; Direct imaging of DNA in living cells reveals the dynamics of chromosome formation. Journal of Cell Biology144:813–821
    [Google Scholar]
  21. Marechal V., Dehee A., Chikhi-Brachet R., Piolot T., Coppey-Moisan M., Nicolas J. C.. 1999; Mapping EBNA-1 domains involved in binding to metaphase chromosomes. Journal of Virology73:4385–4392
    [Google Scholar]
  22. Nonkwelo C., Skinner J., Bell A., Rickinson A., Sample J.. 1996; Transcription start sites downstream of the Epstein–Barr virus (EBV) Fp promoter in early-passage Burkitt lymphoma cells define a fourth promoter for expression of the EBV EBNA-1 protein. Journal of Virology70:623–627
    [Google Scholar]
  23. Petti L., Sample C., Kieff E.. 1990; Subnuclear localization and phosphorylation of Epstein–Barr virus latent infection nuclear proteins. Virology176:563–574
    [Google Scholar]
  24. Piolot T., Tramier M., Coppey M., Nicolas J. C., Marechal V.. 2001; Close but distinct regions of human herpesvirus 8 latency-associated nuclear antigen 1 are responsible for nuclear targeting and binding to human mitotic chromosomes. Journal of Virology75:3948–3959
    [Google Scholar]
  25. Rao P. N.. 1977; Premature chromosome condensation and the fine structure of chromosomes. In Molecular Structure of Human Chromosomes pp205–231 Edited by Yunis J. J.. New York: Academic Press;
    [Google Scholar]
  26. Rickinson A. B., Kieff E.. 2001; Epstein–Barr Virus. In Fields Virology pp2575–2627 Edited by Knipe D. M., Howley P. M.. Philadelphia: Lippincott Williams & Wilkins;
    [Google Scholar]
  27. Sadoni N., Langer S., Fauth C., Bernardi G., Cremer T., Turner B. M., Zink D.. 1999; Nuclear organization of mammalian genomes. Polar chromosome territories build up functionally distinct higher order compartments. Journal of Cell Biology146:1211–1226
    [Google Scholar]
  28. Shire K., Ceccarelli D. F., Avolio-Hunter T. M., Frappier L.. 1999; EBP2, a human protein that interacts with sequences of the Epstein–Barr virus nuclear antigen 1 important for plasmid maintenance. Journal of Virology73:2587–2595
    [Google Scholar]
  29. Sugden B., Warren N.. 1989; A promoter of Epstein–Barr virus that can function during latent infection can be transactivated by EBNA-1, a viral protein required for viral DNA replication during latent infection. Journal of Virology63:2644–2649
    [Google Scholar]
  30. Ward W. W., Bokman S. H.. 1982; Reversible denaturation of Aequorea green-fluorescent protein: physical separation and characterization of the renatured protein. Biochemistry21:4535–4540
    [Google Scholar]
  31. Yates J. L.. 1996; Epstein–Barr virus DNA replication. In DNA Replication in Eukaryotic Cells pp751–774 Edited by DePamphilis M. L.. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  32. Yates J. L., Warren N., Sugden B.. 1985; Stable replication of plasmids derived from Epstein–Barr virus in various mammalian cells. Nature313:812–815
    [Google Scholar]
  33. Yates J. L., Camiolo S. M., Bashaw J. M.. 2000; The minimal replicator of Epstein–Barr virus oriP . Journal of Virology74:4512–4522
    [Google Scholar]
  34. Zhang D., Frappier L., Gibbs E., Hurwitz J., O’Donnell M.. 1998; Human RPA (hSSB) interacts with EBNA1, the latent origin binding protein of Epstein–Barr virus. Nucleic Acids Research26:631–637
    [Google Scholar]
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