1887

Abstract

SUMMARY

Denatured DNA of herpes simplex virus was released from the particles using an alkaline detergent, Decon-75. The largest single strands sedimented on alkaline sucrose gradients with a mol. wt. of 47.2 ± 0.33 × 10, slightly less than half the value calculated for the intact duplex (104 × 10). About 50% of the DNA was found in fragments which sedimented slower than this in a heterogeneous manner. On agarose gel electrophoresis the largest strands migrated with a mol. wt. of 40 × 10. The reason for this difference is not known but since the individual strands of T 4 DNA were shown to migrate with slightly different values, factors other than mol. wt. may affect the migration of single-stranded DNA. Two fragments of mol. wt. 35 × 10 and 30 × 10 were observed but the rest of the fragments remained unresolved by this technique. Virus DNA associated with the nucleus of infected cells had a much lower mol. wt. than particle DNA (3.2 × 10). Although this value increased if the DNA was isolated from intact cells, the average sedimentation coefficient of nuclear virus DNA was never as high as the largest strands of particle DNA.

When the largest single strands from particles were prepared by sucrose gradient fractionation they exhibited a unimodal mol. wt. distribution after both sedimentation and electrophoretic analysis. These ‘intact’ single strands were annealed and analysed by banding in CsCl gradients and by analyses employing the endonuclease. The results show that the ‘intact’ strands reassociated with the same kinetics and to the same extent as total virus DNA, suggesting that both strands of the duplex were present in equal amounts.

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1973-12-01
2024-04-16
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References

  1. Abelson J. N., Thomas C. A. 1966; The anatomy of the T 5 bacteriophage DNA molecule. Journal of Molecular Biology 18:262–291
    [Google Scholar]
  2. Albertsson P. A. 1967; Two phase separation of viruses. In Methods in Virology vol II chap. 19 Edited by Maramorosch K., Koprowski E. New York and London: Academic Press;
    [Google Scholar]
  3. Becker Y., Dym H., Sarov I. 1968; Herpes simplex virus DNA. Virology 36:184–192
    [Google Scholar]
  4. Bollum F. J. 1959; Thermal conversion of nonpriming deoxynucleic acid to primer. Journal of Biological Chemistry 234:2733–2734
    [Google Scholar]
  5. Britten R. J., Kohne D. E. 1968; Repeated sequences in DNA. Science, New York 161:529–540
    [Google Scholar]
  6. Bujard H. 1969; Location of the single strand interruptions in the DNA of phage T 5. Proceedings of the National Academy of Sciences of the United States of America 62:1167–1174
    [Google Scholar]
  7. Burgi E., Hershey A. D. 1963; Sedimentation rate as a measure of molecular weight of DNA. Bio-physical Journal 3:309–321
    [Google Scholar]
  8. Dulbecco R., Vogt M. 1954; Plaque formation and isolation of pure lines with poliomyelitis viruses. Journal of Experimental Medicine 99:167–182
    [Google Scholar]
  9. Fenner F. 1968 In The Biology of Animal Viruses vol 1: chap. 6 New York and London: Academic Press;
    [Google Scholar]
  10. Frenkel N., Roizman B. 1972; Separation of the herpesvirus deoxyribonucleic acid duplex into unique fragments and intact strand on sedimentation in alkaline gradients. Journal of Virology 10:565–572
    [Google Scholar]
  11. Hayward G. S. 1972; Gel electrophoretic separation of the complementary strands of bacteriophage DNA. Virology 49:342–344
    [Google Scholar]
  12. Hayward G. S., Smith M. G. 1972a; The chromosome of bacteriophage T5. I. Analysis of the single- stranded DNA fragments by agarose gel electrophoresis. Journal of Molecular Biology 63:383–395
    [Google Scholar]
  13. Hayward G. S., Smith M. G. 1972b; The chromosome of bacteriophage T5. II. Arrangement of the single- stranded DNA fragments in the T5+ and T5st (0) chromosomes. Journal of Molecular Biology 63:397–407
    [Google Scholar]
  14. Jaquemin-Sablon A., Richardson C. C. 1970; Analysis of the interruptions in bacteriophage T5 DNA. Journal of Molecular Biology 47:477–493
    [Google Scholar]
  15. Kieff E. D., Bachenheimer S. L., Roizmann B. 1971; Size, composition and structure of the deoxyribonucleic acid of subtypes 1 and 2 herpes simplex virus. Journal of Virology 8:125–132
    [Google Scholar]
  16. Kozinski A. W., Szybalski W. 1959; Dispersive transfer of the parental DNA molecule to the progeny of phage Φ × 174. Virology 9:260–274
    [Google Scholar]
  17. Labedan B., Crochet M., Legault-Demare J., Stevens B. J. 1973; Location of the first step transfer fragment and single-strand interruptions in T5 sto bacteriophage DNA. Journal of Molecular Biology 75:213–234
    [Google Scholar]
  18. Lanni Y. T. 1961; Invasion by bacteriophage T 5. III. Stages revealed by changes in susceptibility of early complexes to abortive infection. Virology 15:127–135
    [Google Scholar]
  19. Lee L., Kieff E. D., Bachenheimer S., Roizman B. 1971; The size and composition of Marek′s disease virus DNA. Journal of Virology 72:289–294
    [Google Scholar]
  20. Linn S., Lehman I. R. 1965; An endonuclease from Neurospora Crassa specific for polynucleotides lacking an ordered structure. Journal of Biological Chemistry 240:1287–1293
    [Google Scholar]
  21. Luria S. E., Darnell J. E. 1968 In General Virology 2nd ed pp 117 New York, London, Sydney: John Wiley;
    [Google Scholar]
  22. Macpherson I., Stoker M. 1962; Polyoma transformation of hamster cell clones-an investigation of genetic factors affecting cell competence. Virology 16:147
    [Google Scholar]
  23. McHattie L. A., Ritchie D. A., Thomas C. A., Richardson C. C. 1967; Terminal repetition in permuted T 2 bacteriophage DNA molecules. Journal of Molecular Biology 23:355–363
    [Google Scholar]
  24. Reznikoff W. S., Thomas C. A. 1969; The anatomy of the SP 50 bacteriophage DNA molecule. Virology 37:309–317
    [Google Scholar]
  25. Ritchie D. A., Malcolm F. E. 1970; Heat stable and density mutants of phages T 1, T 3 and T 7. Journal of General Virology 9:35–43
    [Google Scholar]
  26. Ritchie D. A. 1970; Physical characterization of the DNA released from phage T 4 particles by heat inactivation. FEBS Letters II:257–260
    [Google Scholar]
  27. Russell W. C. 1962; A sensitive and precise plaque assay for herpes virus. Nature, London 195:1028–1029
    [Google Scholar]
  28. Russell W. C., Gold E., Keir H. M., Omura H., Watson D. H., Wildy P. 1964; The growth of herpes simplex virus and its nucleic acid. Virology 22:103–110
    [Google Scholar]
  29. Schildkraut C. L., Marmur J., Doty P. 1962; Determination of the base composition of deoxynucleic acid from its buoyant density in CsCl. Journal of Molecular Biology 4:430–443
    [Google Scholar]
  30. Szybalski W. 1968 In Equilibrium Sedimentation of Viruses, Nucleic Acids and other Macromolecules in Density Gradients Fractions no. 1 Palo Alto, U,. S. A.: Spinco Division of Beckman Instruments Inc;
    [Google Scholar]
  31. Thomas C. A., Mchattie L. A. 1964; Circular T 2 DNA molecules. Proceedings of the National Academy of Sciences of the United States of America 52:1297–1301
    [Google Scholar]
  32. Thomas C. A., Mchattie L. A. 1967; The anatomy of viral DNA molecules. Annual Review of Biochemistry 36: (II) 485–518
    [Google Scholar]
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