1887

Abstract

SUMMARY

Epstein–Barr virus (EBV) nuclear antigen (EBNA) was purified from the Burkitt lymphoma line Raji and its EBV DNA-binding properties were characterized. EBNA binding protected fragments of about 30 bp of B95-8 cell-derived EBV DNA from an excess of DNase I. Human anti-EBNA antibodies prevented DNA binding. Purified extracts from EBNA-negative cells did not protect EBV DNA against DNase I digestion. Mapping of the EBV DNA fragments protected from endonuclease (RI, dIII, I) digestion revealed many binding sites. Similar results were obtained following mixing of crude cell extracts and dIII-digested fragments of EBV DNA and subsequent immunoprecipitation of the EBNA-DNA complex. In experiments involving the analysis of EBV DNA, fragments were protected from DNase I digestion by purified EBNA.

Keyword(s): DNA-binding protein , DNase I and EBNA
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/content/journal/jgv/10.1099/0022-1317-65-11-2043
1984-11-01
2022-01-29
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References

  1. Bornkamm G. W., Delius H., Zimber U., Hudewetz J., Epstein M. A. 1980; Comparison of Epstein-Barr virus strains of different origin by analysis of viral DNAs. Journal of Virology 35:603–618
    [Google Scholar]
  2. Chandler D. W., Gralla J. 1980; Specific binding and protection of form II SV40 deoxyribonucleic acid by ribonucleic acid polymerase II from wheat germ. Biochemistry 19:1604–1612
    [Google Scholar]
  3. Hennessy K., Heller M., Van Santen V., Kieff E. 1983; Simple repeat array in Epstein-Barr virus DNA encodes part of the Epstein-Barr nuclear antigen. Science 220:1396–1398
    [Google Scholar]
  4. Hirsch I., Suchankova A., Zavadova H., Vonka V. 1978; Study of Epstein-Barr virus determined nuclear antigen (EBNA) by chromatography on fixed cell nuclei. International Journal of Cancer 22:535–541
    [Google Scholar]
  5. Luka J., Siegert W., Klein G. 1977; Solubilization of the Epstein-Barr virus-determined nuclear antigen and its characterization as a DNA-binding protein. Journal of Virology 22:1–8
    [Google Scholar]
  6. Luka J., Lindahl T., Klein G. 1978; Purification of the Epstein-Barr virus-determined nuclear antigen from Epstein-Barr virus-transformed human lymphoid cell lines. Journal of Virology 27:604–611
    [Google Scholar]
  7. Luka J., Jornval H., Klein G. 1980; Purification and biochemical characterization of the Epstein-Barr virus-determined nuclear antigen and an associated protein with a 53000 dalton subunit. Journal of Virology 35:592–602
    [Google Scholar]
  8. Luka J., Jornvall H., Klein G. 1983; The Epstein-Barr virus determined nuclear antigen (EBNA): a previously identified 48K component and higher molecular weight forms of the antigen are structurally related. Intervirology 20:213–222
    [Google Scholar]
  9. Maxam A. M., Gilbert W. 1977; A new method for sequencing DNA. Proceedings of the National Academy of Sciences, U. S. A 74:560–564
    [Google Scholar]
  10. Ohno S., Luka J., Lindahl T., Klein G. 1977; Identification of a purified complement-fixing antigen as Epstein-Barr virus-determined nuclear antigen (EBNA) by its binding to metaphase chromosomes. Proceedings of the National Academy of Sciences, U. S. A 74:1605–1609
    [Google Scholar]
  11. Rigby P. W. J., Dieckmann M., Rhodes C., Berg P. 1977; Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  12. Scheller A., Covey L., Barnet B., Prlves C. 1982; A small subclass of SV40 T antigen binds to the viral origin of replication. Cell 29:375–383
    [Google Scholar]
  13. Smith G. E., Summers M. D. 1980; The bidirectional transfer of DNA and RNA to nitrocellulose or diazo-benzyloxymethyl-paper. Analytical Biochemistry 109:123–129
    [Google Scholar]
  14. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  15. Strnad B. C., Schuster B. C., Hopkins R. F., Neubauer R. H., Robin H. 1981; Identification of an Epstein-Barr virus nuclear antigen by fluoroimmunoelectrophoresis and radioimmunoelectrophoresis. Journal of Virology 38:996–1004
    [Google Scholar]
  16. Summers W. P., Crogan E. A., Shedd D., Robert M., Liu Chun-Ren., Miller G. 1982; Stable expression in mouse cells of nuclear neoantigen after transfer of a 3.4-megadalton cloned fragment of Epstein-Barr virus DNA. Proceedings of the National Academy of Sciences, U. S. A 79:5688–5692
    [Google Scholar]
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