1887

Abstract

SUMMARY

The fate of parental SV40 DNA in monkey kidney cells was investigated by infecting the cells with purified virus labelled with [H]-thymidine. 5-Bromodeoxyuridine (200 µg/ml) was added to the cells at 2 h after infection to label virus progeny DNA. At 72 h post-infection the cells were harvested and virus DNA was extracted and fractionated by isopycnic sedimentation in CsCl solution. The following DNAs with characteristic densities were found: light (LL), 1.70 g/ml; hybrid (HL), 1.75 g/ml and heavy DNA (HH), 1.80 g/ml.

Of the total cell-associated [H]-radioactivity (derived from parental virus), more than 90% was recovered in unreplicated parental DNA (LL DNA), about 2% was recovered in the HL DNA and about 0.7% was associated with the HH DNA. The unreplicated parental DNA was present as uncoated intact DNA complexed with proteins present in the infected cell. The HL DNA contains one light parental and one heavy progeny DNA strand. The nature of the radioactivity present in the HH DNA remains to be determined.

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1975-04-01
2022-01-27
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References

  1. Barbanti’Brodano G., Swetly P., Koprowski H. 1970; Early events in the infection of permissive cells with simian virus 40: adsorption, penetration, and uncoating. Journal of Virology 6:78–86
    [Google Scholar]
  2. Berkowitz D. M., Kakefunda T., Sporn M. B. 1969; A simple and rapid method for the isolation of enzymatically active HeLa cell nuclei. Journal of Ceil Biology 42:851–855
    [Google Scholar]
  3. Black P. H., Crawford E. M., Crawford L. V. 1964; The purification of simian virus 40. Virology 24:381–387
    [Google Scholar]
  4. Bourgaux P., Bourgaux-Ramoisy D., Dulbecco R. 1969; The replication of the ring-shaped DNA of polyoma virus. I. Identification of the replicative intermediate. Proceedings of the National Academy of Sciences of the United States of America 64:701–708
    [Google Scholar]
  5. Bourgaux P., Bourgaux-Ramoisy D. 1972; Unwinding of replicating polyoma virus DNA. Journal of Molecular Biology 70:399–413
    [Google Scholar]
  6. Calothy G., Hirai K., Defendi V. 1973; 5-Bromodeoxyuridine incorporation into simian virus 40 deoxyribonucleic acid. Elfects on simian virus 40 replication in monkey cells. Virology 55:329–338
    [Google Scholar]
  7. Crawford L. V., Black P. H. 1964; The nucleic acid of simian virus 40. Virology 24:388–392
    [Google Scholar]
  8. Dubbs D. R., Rachmeler M., Kit S. 1974; Recombination between temperature-sensitive mutants of simian virus 40. Virology 57:161–174
    [Google Scholar]
  9. Green M. H., Miller H. I., Hendler S. 1971; Isolation of a polyoma-nucleoprotein complex from infected mouse-cell cultures. Proceedings of the National Academy of Sciences of the United States of America 68:1032–1036
    [Google Scholar]
  10. Hall M. R., Meinke W., Goldstein D. A. 1973; Nucleoprotein complexes containing replicating simian virus 40 DNA: comparison with polyoma nucleoprotein complexes. Journal of Virology 12:901–908
    [Google Scholar]
  11. Hirai K., Defendi V. 1972; Integration of simian virus 40 deoxyribonucleic acid into the deoxyribonucleic acid of permissive monkey kidney cells. Journal of Virology 9:705–707
    [Google Scholar]
  12. Hirt B. 1966; Evidence for semi-conservative replication of circular polyoma DNA. Proceedings of the National Academy of Sciences of the United States of America 55:997–1004
    [Google Scholar]
  13. Hirt B. 1967; Selective extraction of polyoma DNA from infected mouse cell cultures. Journal of Molecular Biology 26:365–369
    [Google Scholar]
  14. Hirt B. 1969; Replicating molecules of polyoma virus DNA. Journal of Molecular Biology 40:141–144
    [Google Scholar]
  15. Holloway P. W. 1973; A simple procedure for removal of Triton X-100 from protein samples. Analytical Biochemistry 53:304–308
    [Google Scholar]
  16. Holzel F., Sokol F. 1974; Integration of progeny simian virus 40 DNA into the host cell genome. Journal of Molecular Biology 84:423–444
    [Google Scholar]
  17. Hummeler K., Tomassini N., Sokol F. 1970; Morphological aspects of the uptake of simian virus 40 by permissive cells. Journal of Virology 6:87–93
    [Google Scholar]
  18. Jaenisch R., Mai’er A., Levine A. 1971; Replicating SV40 molecules containing closed circular template DNA strands. Nature New Biology 233:72–75
    [Google Scholar]
  19. Kozinski A. W., Kozinski P. B. 1963; Fragmentary transfer of 32P-labeled parental DNA to progeny phage. II, The average size of the transferred parental fragment. Two-cycle transfer. Repair of the polynucleotide chain after fragmentation. Virology 20:213–229
    [Google Scholar]
  20. Levine A. J., Kang H. S., Billheimer F. E. 1970; DNA replication in SV40 infected cells. I. Analysis of replicating SV40 DNA. Journal of Molecular Biology 50:549–568
    [Google Scholar]
  21. Radloff R., Bauer W., Vinograd J. 1967; A dye-buoyant-density method for the detection 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]
  22. Ritzi E. M., Levine A. J. 1973; The fragmentation of cellular DNA and the formation of pseudovirions during SV40 infection of African green monkey kidney cells. Journal of General Virology 20:353–367
    [Google Scholar]
  23. Sebring E. D., Kelly T. J. Jun., Thoren M. M., Salzman N. P. 1971; Structure of replicating simian virus 40 deoxyribonucleic acid molecules. Journal of Virology 8:478–490
    [Google Scholar]
  24. Shmookler R. J., Buss J., Green M. H. 1974; Properties of the polyoma virus transcription complex obtained from mouse nuclei. Virology 57:122–127
    [Google Scholar]
  25. Sokol F., Huebner F. K., Tan K. B., Sheehan M. C., Santoli D. 1974; Heterogeneity of simian virus 40 population: virus particles containing one or two virus DN A molecules?. Journal of General Virology 22:55–71
    [Google Scholar]
  26. Tan K. B., Sokol F. 1972; Structural proteins of simian virus 40: phosphoproteins. Journal of Virology 10:985–994
    [Google Scholar]
  27. Tan K. B., Sokol F. 1973; Phosphorylation of simian virus 40 proteins in a cell-free system. Journal of Virology 12:696–703
    [Google Scholar]
  28. Tan K. B., Sokol F. 1974; Replication of simian virus 40 in permissive cells: assembly of virus components. Journal of General Virology 25:37–51
    [Google Scholar]
  29. Wake R. G., Baldwin R. L. 1962; Physical studies on the replication of DNA in vitro. Journal of Molecular Biology 5:201–216
    [Google Scholar]
  30. White M., Eason R. 1971; Nucleoprotein complexes in simian virus 40-infected cells. Journal of Virology 8363–371
    [Google Scholar]
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