1887

Abstract

Summary

An early temperature-sensitive event which prevents the replication of variola-virus DNA in HeLa cells at 40° was studied in ‘temperature-shift’ experiments using 5-fluorodeoxyuridine or 5-bromodeoxyuridine. ‘Shift-up’ from 35° to 40° suppressed the replication of virus DNA, rapidly halting it even after it had begun at 35°. ‘Shift-down’ from 40° to 35° permitted the replication of virus DNA after a delay of not more than 2 hr. These results were confirmed in studies of the incorporation of [H]thymidine into DNA in the cytoplasmic fraction. It was also shown that the incorporation which followed ‘shift-down’ from 40° could be inhibited by adding -fluorophenylalanine at the time of shift. When cytoplasmic fractions from cells incubated at different temperatures were examined for enzyme activities, there was a marked increase in thymidine kinase activity in infected cells both at 35° and at 40°. DNA polymerase activity was increased five- to sixfold in infected cells at 35° but was not increased at 40° or in the presence of -fluorophenylalanine. The increased polymerase activity in infected cells at 35° was unstable at 40° both and , in contrast to its greater stability at 35°. The behaviour of this enzyme was thought to explain the observed temperature-sensitivity of the replication of variola virus DNA.

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/content/journal/jgv/10.1099/0022-1317-4-3-413
1969-04-01
2022-01-24
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References

  1. Bedson H. S., Cruickshank J. G. 1968; Multiple protein functions in the replication of pox virus DNA. J. gen. Virol 3:147
    [Google Scholar]
  2. Chrispeels M. J., Boyd R. F., Williams L.-S., Neidhardt F. C. 1968; Modification of valyl-t-RNA synthetase by bacteriophage in Escherichia coli. J. molec. Biol 31:463
    [Google Scholar]
  3. Cohen S. S., Flaks J. G., Barner H. D., Loeb M. R., Lichtenstein J. 1958; The mode of action of 5 fluorouracil and its derivatives. Proc. natn. Acad. Sci. U.S.A. 44:1004
    [Google Scholar]
  4. Cruickshank J. G., Bedson H. S. 1968; Temperature-sensitive events in the growth of variola virus in HeLa cells. J. gen. Virol 3:255
    [Google Scholar]
  5. De Waard A., Paul A. V., Lehman I. R. 1965; The structural gene for deoxyribonucleic acid polymerase in bacteriophages T 4 and T 5. Proc. natn. Acad. Sci. U.S.A. 54:1241
    [Google Scholar]
  6. Dubbs D. R., Kit S. 1964; Isolation and properties of vaccinia mutants deficient in thymidine kinase inducing activity. Virology 32:214
    [Google Scholar]
  7. Easterbrook K. B., Davern C. I. 1963; The effect of 5-bromodeoxyuridine in the multiplication of vaccinia virus. Virology 19:509
    [Google Scholar]
  8. Fenner F., Sambrook J. F. 1964; The genetics of animal viruses. Ann. Rev. Microbiol 18:47
    [Google Scholar]
  9. Joklik W. K. 1966; The poxviruses. Bact. Rev 30:33
    [Google Scholar]
  10. Joklik W. K., Becker Y. 1964; The replication and coating of vaccinia DNA. J. molec. Biol 10:452
    [Google Scholar]
  11. Jungwirth C., Joklik W. K. 1965; Studies on ‘early’ enzymes in HeLa cells infected with vaccinia virus. Virology 27:80
    [Google Scholar]
  12. Jungwirth C., Launer J. 1968; Effect of poxvirus infection on host cell deoxyribonucleic acid synthesis. J. Virol 2:401
    [Google Scholar]
  13. Kates J. R., McAuslan B. R. 1967; Relationship between protein synthesis and viral deoxyribonucleic acidsynthesis. J. Virol 1:110
    [Google Scholar]
  14. Keir H. M., Shepherd J. B. 1965; Thiol groups in deoxyribonucleic acid nucleotidyltransferase. Biochem. J 95:483
    [Google Scholar]
  15. Klemperer H. G., Haynes G. R., Shedden W. I. H., Watson D. H. 1967; A virus-specific thymidine kinase in BHK 2I cells infected with herpes simplex virus. Virology 31:120
    [Google Scholar]
  16. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folinphenol reagent. J. biol. Chem 93:265
    [Google Scholar]
  17. Magee W. E., Miller O. V. 1967; Immunological evidence for the appearance of a new DNA-polymerase in cells infected with vaccinia virus. Virology 31:64
    [Google Scholar]
  18. Regan D. J., Chu E. M. Y. 1966; A convenient method for assay of DNA synthesis in synchronised human cell cultures. J. cell. Biol 28:139
    [Google Scholar]
  19. Salzman N. P. 1960; The rate of formation of vaccinia deoxyribonucleic acid and vaccinia virus. Virology 10:150
    [Google Scholar]
  20. Stevens J. G. 1966; Selective inhibition of herpes virus DNA synthesis at elevated temperature. Virology 29:570
    [Google Scholar]
  21. Stevens J. G., Jackson N. L. 1967; Studies on a temperature-sensitive step essential to herpesvirus DNA replication. Virology 32:654
    [Google Scholar]
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