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1887

Abstract

SUMMARY

Equilibrium density gradient centrifugation in CsCl confirms that DNA synthesized after vaccinia virus infection of HeLa cells is homogeneous in buoyant density and thus in base composition and is similar in this respect to bulk HeLa cell DNA. In contrast, rate sedimentation in alkaline sucrose gradients distinguishes two main classes of virus-induced DNA, neither of which can be equated with cell DNA synthesized in the same cultures prior to infection. The slower sedimenting class of virus-induced DNA co-sediments with DNA from purified virus particles: the second class sediments faster than pre-labelled cell DNA. Heterogeneity of virus-induced DNA does not result from fragmentation of radioactively labelled DNA, virus-mediated breakdown of cell DNA or association with either proteins or polyamines. Both slow and fast sedimenting classes of virus-induced DNA contain sequences complementary to all restriction endonuclease III-specific fragments of the virus genome. The multiple species of DNA synthesized after infection are distinguished further by the effect of ethidium bromide. At a concentration which prevents the formation of infectious progeny virus, this compound inhibits selectively the synthesis of that class of virus-induced DNA which sediments faster in alkaline sucrose gradients.

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/content/journal/jgv/10.1099/0022-1317-42-1-223
1979-01-01
2025-01-21
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References

  1. Archard L. C., Williamson J. D. 1971; The effect of arginine deprivation on the replication of vaccinia virus. Journal of General Virology 12:249–258
    [Google Scholar]
  2. Berns K. I., Silverman C. 1970; Natural occurrence of cross-linked vaccinia virus deoxyribonucleic acid. Journal of Virology 5:299–304
    [Google Scholar]
  3. Cairns J. 1960; The initiation of vaccinia infection. Virology 11:603–634
    [Google Scholar]
  4. Dahl R., Kates J. R. 1970a; Intracellular structures containing vaccinia DNA: isolation and characterization. Virology 42:453–462
    [Google Scholar]
  5. Dahl R., Kates J. R. 1970b; Synthesis of vaccinia virus ‘early’ and ‘late’ messenger RNA in vitro with nucleoprotein structures isolated from infected cells. Virology 42:463–472
    [Google Scholar]
  6. Geshelin P., Berns K. I. 1974; Characterization and localisation of the naturally occurring cross-links in vaccinia virus DNA. Journal of Molecular Biology 88:785–796
    [Google Scholar]
  7. Harford C. G., Hamlin A., Rieders E. 1966; Electron microscopic autoradiography of DNA synthesis in cells infected with vaccinia virus. Experimental Cell Research 42:50–57
    [Google Scholar]
  8. Jeffreys A. J., Flavell R. A. 1977; A physical map of the DNA regions flanking the rabbit β-globin gene. Cell 12:429–439
    [Google Scholar]
  9. Joklk W. K. 1962; The purification of four strains of poxvirus. Virology 18:9–18
    [Google Scholar]
  10. Joklik W. K., Becker Y. 1964; The replication and coating of vaccinia DNA. Journal of Molecular 1Biology 10:452–474
    [Google Scholar]
  11. Jungwirth C., Dawid I. B. 1967; Vaccinia DNA: separation of virus from host cell DNA. Archiv fur die gesamte Virusforschung 20:464–468
    [Google Scholar]
  12. Loh P. C., Riggs J. L. 1961; Demonstration of the sequential development of vaccinial antigens and virus in infected cells: observations with cytochemical and differential fluorescent proceedures. Journal of Experimental Medicine 114:149–160
    [Google Scholar]
  13. Muller H. K., Witter R., Schaffner W., Schümperli D., Menna A., Wyler R. 1978; Comparison of five poxvirus genomes by analysis with restriction endonucleases Hind III, Bam I and EcoRI. Journal of General Virology 38:135–147
    [Google Scholar]
  14. Parkhurst J. R., Peterson A. R., Heidelberger C. 1973; Breakdown of HeLa cell DNA mediated by vaccinia virus. Proceedings of the National Academy of Sciences of the United States of America 70:3200–3204
    [Google Scholar]
  15. Peterson A. R., Fox B. W. 1971; Toxicity of tritiated thymidine in P388F lymphoma cells. II. Effects on DNA molecular weight. International Journal of Radiation Biology 19:237–246
    [Google Scholar]
  16. Pogo B. G. T., Dales S. 1974; Biogenesis of poxviruses: further evidence for inhibition of host and virus DNA synthesis by a component of the invading inoculum particle. Virology 58:377–386
    [Google Scholar]
  17. Polisky B., Kates J. 1972; Vaccinia virus intracellular DNA-protein complex: biochemical characteristics of associated protein. Virology 49:168–179
    [Google Scholar]
  18. Radloff R., Bauer W., Vinograd J. 1967; A dye-buoyant-density method for the selection and isolation of closed circular duplex DNA: The closed circular DNA in HeLa cells. Proceedings of the National Academy of Sciences of the United States of America 57:1514–1521
    [Google Scholar]
  19. Raya J. M., Vilagines R. 1970; Action du bromhydrate d’éthidium sur le développement du virus vaccinal dans les cellules en culture de rein de babouin. II. Action sur les syntheses d’ADN et d’ARN de cellules infectées et non infectées. Archiv fur die gesamte Virusforschung 30:155–158
    [Google Scholar]
  20. Rigby P. W. J., Dieckmann M., Rhodes C., Berg P. 1977; Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  21. Sarov I., Becker Y. 1967; Studies on vaccinia virus DNA. Virology 33:369–375
    [Google Scholar]
  22. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–518
    [Google Scholar]
  23. Studier F. W. 1965; Sedimentation studies of the size and shape of DNA. Journal of Molecular Biology n:373–390
    [Google Scholar]
  24. Vilagines R. 1970; Action du bromohydrate d’éthidium sur le développement du virus vaccinial dans les cellules en culture de rein de babouin. I. Action sur la cinétique de développement viral. Archiv für die gesamte Virusforschung 30:59–66
    [Google Scholar]
  25. Williamson J. D., Archard L. C. 1976; The effect of canaline on some events in vaccinia virus replication. Journal of General Virology 30:81–89
    [Google Scholar]
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