1887

Abstract

SUMMARY

The nature of the DNA molecules synthesized in nuclei of herpes simplex virus (HSV)-infected cells and was studied by chromatography on BND-cellulose columns after shearing to DNA fragments of 10 to 20 × 10 daltons. The incorporation of labelled precursors occurs in the DNA fragments containing single-stranded regions, presumably the replication forks. Prolongation of DNA synthesis leads to the accumulation of labelled DNA fragments that lack single-stranded sequences. Analysis of the isolated DNA fragments by density centrifugation in CsCl gradients revealed that most of the labelled DNA molecules are of virus specificity and the minority are cellular DNA fragments. Double-stranded virus DNA fragments and virus DNA fragments containing single-stranded sequences band in CsCl gradients at a density of 1.718 g/ml, the density of virion DNA. This suggests that the replicating HSV DNA molecules have the same density as the virion DNA and contain relatively little single-stranded DNA. The synthesis of HSV DNA molecules under conditions in isolated nuclei occurs by incorporation of the precursors into DNA fragments with single-stranded regions. The synthesis of cellular DNA in nuclei from hydroxyurea and cytosine arabinoside treated cells also occurs by elongation of nascent DNA chains.

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1976-08-01
2024-10-12
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References

  1. BECKER Y., ASHER Y. 1975; [i]In vitro[/i] synthesis of DNA in nuclei isolated from herpes simplex virus-infected cells, untreated and treated with metabolic inhibitors. Virology 63:209–220
    [Google Scholar]
  2. BECKER Y., OLSHEVSKY U., LEVITT J. 1967; The role of arginine in the replication of herpes simplex virus. Journal of General Virology i:471–478
    [Google Scholar]
  3. BEN-ZEEV A., WEINBERG E., BECKER Y. 1974; Isolation of herpes simplex virus DNA from virus particles and infected cells by electrophoresis in Polyacrylamide gels. Journal of General Virology 25:63–73
    [Google Scholar]
  4. GORDIN M., OLSHEVSKY U., ROSENKRANZ H. S., BECKER Y. 1973; Studies on herpes simplex virus DNA: denaturation properties. Virology 55:280–284
    [Google Scholar]
  5. HIGGINS N. P., KATO K., STRAUSS B. 1976; A model for replication repair in mammalian cells. Cancer Research In the press
    [Google Scholar]
  6. KIGER J. A., SINSHEIMER R. L. 1969; Vegetative lambda DNA. IV. Fractionation of replicating lambda DNA on benzoylated-naphthoylated DEAE cellulose. Journal of Molecular Biology 40:467–490
    [Google Scholar]
  7. KLEINSCHMIDT A. K., LANG D., JACHERTS D., ZAHN R. K. 1962; Darstellung und langenmessungen des gesamten desoxyribonucleinsäure-inhaltes von T[sub]2[/sub]-bakteriophagen. Biochimica et Biophysica Acta 61:857–864
    [Google Scholar]
  8. KLEINSCHMIDT A. K., DUNNEBAKE T. H., SPENDLOVE R. S., SCHAFFER F. L., WHITCOMB R. F. 1964; Electron microscopy of RNA from reovirus and wound tumor virus. Journal of Molecular Biology 10:282–288
    [Google Scholar]
  9. LEVITT J., BECKER Y. 1967; The effect of cytosine arabinoside on the replication of herpes simplex virus. Virology 31:129–134
    [Google Scholar]
  10. ROSENKRANZ H. S., BECKER Y. 1973; Reversible inhibition of herpes simplex virus replication by hydroxyurea. Antimicrobial Agents and Chemotherapy 3:325–331
    [Google Scholar]
  11. SCUDIERO D., HENDERSON E., NORIN A., STRAUSS B. 1975; The measurement of chemically-induced DNA repair synthesis in human cells by BND-cellulose chromatography. Mutation Research 29:473–488
    [Google Scholar]
  12. SCUDIERO D., STRAUSS B. 1974; Accumulation of single-stranded regions in DNA and the block to replication in a human cell line alkylated with methyl methane sulfonate. Journal of Molecular Biology 83:17–34
    [Google Scholar]
  13. SHLOMAI J., FRIEDMANN A., BECKER Y. 1976; Replicative intermediates of herpes simplex virus DNA. Virology 69:647–659
    [Google Scholar]
  14. SUSSENBACH J. S., VAN DER VLIET P. C, ELLENS D. J., JANSZ H. S. 1972; Linear intermediates in the replication of adenovirus DNA. Nature New Biology 239:47–49
    [Google Scholar]
  15. WEISSBACH A., HONG S. C, AUCKER J., MULLER R. 1973; Characterization of herpes simplex virus-induced deoxyribonucleic acid polymerase. Journal of Biological Chemistry 248:6270–6277
    [Google Scholar]
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