1887

Abstract

SUMMARY

Reovirus infection inhibits the incorporation of H-thymidine into cellular DNA. We have now investigated several aspects of this inhibition in L-929 cells early (8 h) after infection at high multiplicity (200 to 250 p.f.u./cell). Using equilibrium sedimentation analysis of DNA sequentially labelled with density and radioactive analogues of thymidine, we find a 52% reduction in the amount of DNA synthesized with no change in rate of replication fork movement in infected cells. Gel electrophoresis of histones labelled with H-lysine shows that infection inhibits their synthesis by 76% several hours before overall cellular protein synthesis is inhibited. There is also a reduction of nearly 50% in the size of the thymidine triphosphate pool, as measured by enzymic assay. The proportion of exogenous nucleotide in the pool is the same as in uninfected cells since there is no change in the buoyant density of DNA labelled during a short pulse with H-bromodeoxyuridine. The uptake of thymidine is reduced, but its phosphorylation to thymidine triphosphate is normal. The findings provide direct evidence that DNA synthesis is inhibited early in infection. This inhibition is accompanied by other derangements of thymidine and chromatin metabolism suggesting that there is an early and specific attack by reovirus on nuclear function in infected cells.

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1978-06-01
2022-06-27
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References

  1. Britten R. J., Graham D. E., Neufield B. R. 1974 In Methods in Enzymology vol 29 chapter 29 pp 363–408 Edited by Grossman L., Moldave R. New York: Academic Press;
    [Google Scholar]
  2. Brownstein B. L., Rozengurt E., De Asua L. J., Stoker M. 1975; Dissociation by cytochalasin B of movement, DNA synthesis and transport in 3T3 cells. Journal of Cellular Physiology 85:579–586
    [Google Scholar]
  3. Cox D. C., Clinkscales C. W. 1976; Infectious reovirus subviral particles: virus replication, cellular cytopathology, and DNA synthesis. Virology 74:259–261
    [Google Scholar]
  4. Cox D. C., Shaw J. E. 1970; Reovirus alteration of cellular DNA synthesis. Annals of the Oklahoma Academy of Sciences 1:28–37
    [Google Scholar]
  5. Cox D. C., Shaw J. E. 1974; Inhibition of the initiation of cellular DNA synthesis after reovirus infection. Journal of Virology 13:760–761
    [Google Scholar]
  6. Elgin S. C. R., Weintraub H. 1975; Chromosomal proteins and chromatin structure. Annual Review of Biochemistry 44:725–774
    [Google Scholar]
  7. Ensminger W. D. 1969; Viral inhibition of mammalian cell DNA synthesis. Ph. D. thesis Rockefeller University; U. S. A.:
    [Google Scholar]
  8. Ensminger W. D., Tamm I. 1969a; Cellular DNA and protein synthesis in reovirus-infected L cells. Virology 39:358–360
    [Google Scholar]
  9. Ensminger W. D., Tamm I. 1969b; The step in cellular DNA synthesis blocked by reovirus infection. Virology 39:935–938
    [Google Scholar]
  10. Ensminger W. D., Tamm I. 1970; Inhibition of synchronized cellular deoxyribonucleic acid synthesis during Newcastle disease virus, mengovirus or reovirus infection. Journal of Virology 5:672–676
    [Google Scholar]
  11. Follett E. A. C., Pringle C. R., Pennington T. H. 1975; Virus development in enucleate cells: echovirus, poliovirus, pseudorabies virus, reovirus, respiratory syncytial virus and Semliki Forest virus. Journal of General Virology 26:183–196
    [Google Scholar]
  12. Gautschi J. R., Kern R. M. 1973; DNA replication in mammalian cells in the presence of cycloheximide. Experimental Cell Research 80:15–27
    [Google Scholar]
  13. Gomatus P. J., Tamm I. 1963; Macromolecular synthesis in reovirus-infected cells. Biochimica et Bio-physica Acta 72:651–653
    [Google Scholar]
  14. Grunicke H., Hirsch F., Wolf H., Bauer V., Kieffer G. 1975; Selective inhibition of thymidine transport at low doses of the alkylating agent triethylene iminobenzoquinone (Trenimon). Experimental Cell Research 90:357–364
    [Google Scholar]
  15. Hand R. 1975; DNA replication in mammalian cells: altered patterns of initiation during inhibition of protein synthesis. Journal of Cell Biology 67:761–774
    [Google Scholar]
  16. Hand R. 1976a; Thymidine metabolism and DNA synthesis in Newcastle disease virus-infected cells. Journal of Virology 19:801–809
    [Google Scholar]
  17. Hand R. 1976b; Chromosomal DNA replication: retarded fork progression and altered initiation in cells infected with mengovirus or Newcastle disease virus. Virology 81:609–614
    [Google Scholar]
  18. Hand R., Oblin C. 1977; DNA synthesis in mengovirus-infected cells: mechanism of inhibition. Journal of General Virology 37:349–358
    [Google Scholar]
  19. Hand R., Tamm I. 1971; Reovirus: analysis of proteins from released and cell-associated virus. Journal of General Virology 12:121–130
    [Google Scholar]
  20. Hand R., Tamm L. 1972; Rate of DNA chain growth in mammalian cells infected with cytocidal RNA viruses. Virology 47:331–337
    [Google Scholar]
  21. Hand R., Tamm L. 1974; Initiation of DNA replication in mammalian cells and its inhibition by reovirus infection. Journal of Molecular Biology 82:175–183
    [Google Scholar]
  22. Hand R., Tamm I. 1977; Inhibition of mammalian DNA replication by dichlorobenzimidazole riboside. Experimental Cell Research 107:343–354
    [Google Scholar]
  23. Hauschka P. V. 1973 In Methods in Cell Biology vol 7 chapter 19 pp 362–462 Edited by Prescott D. M. New York: Academic Press;
    [Google Scholar]
  24. Hauschka P. V., Everhart L. P., Rubin R. W. 1972; Alteration of nucleoside transport of Chinese hamster cells by dibutyryl adenosine 3′:5′-cyclic monophosphate. Proceedings of the National Academy of Sciences of the United States of America 69:3542–3546
    [Google Scholar]
  25. Joklik W. 1974 In Comprehensive Virology vol 2 chapter 5 pp 231–334 Edited by FraenkelConrat H., Wagner R. New York: Plenum Press;
    [Google Scholar]
  26. Laemmli U. K. 1970; Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature, London 237:680–685
    [Google Scholar]
  27. Lindberg U., Skoog L. 1970; A method for the determination of dATP and dTTP in picomole amounts. Analytical Biochemistry 30:152–160
    [Google Scholar]
  28. Lowry O. G., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  29. Margolis G., Kilham L., Baringer J. R. 1975; Identity of Cowdry type B inclusions and nuclear bodies: observations in reovirus encephalitis. Experimental and Molecular Pathology 23:228–244
    [Google Scholar]
  30. Meuth M., Green H. 1974; Induction of a deoxycytidineless state in cultured mammalian cells by bromo-deoxyuridine. Cell 2:109–112
    [Google Scholar]
  31. Painter R. B., Schaefer A. W. 1969; Rate of synthesis along replicons of different kinds of mammalian cells. Journal of Molecular Biology 45:467–479
    [Google Scholar]
  32. Plagemann P. G. W. 1971; Nucleotide pools of Novikoff rat hepatoma cells growing in suspension culture. 1. Kinetics of incorporation of nucleosides into nucleotide pools and pool sizes during growth cycle. Journal of Cellular Physiology 77:213–240
    [Google Scholar]
  33. Shatkin A. J. 1965; Actinomycin and the differential synthesis of reovirus and L-cell RNA. Biochemical and Biophysical Research Communications 19:506–510
    [Google Scholar]
  34. Shaw J. E., Cox D. C. 1973; Early inhibition of cellular DNA synthesis by high multiplicities of infectious and UV-inactivated reovirus. Journal of Virology 12:704–710
    [Google Scholar]
  35. Silverstein S. C., Astell C., Christman J., Klett H., Acs G. 1974; Synthesis of reovirus oligo adenylic acid in vivo and in vitro. Journal of Virology 13:740–752
    [Google Scholar]
  36. Solter A. W., Hanschumacher R. E. 1969; A rapid quantitative determination of deoxyribonucleoside triphosphates based on the enzymatic synthesis of DNA. Biochemica et Biophysica Acta 174:585–590
    [Google Scholar]
  37. Weintraub H., Holtzer H. 1972; Fine control of DNA synthesis in developing chick red blood cells. Journal of Molecular Biology 66:13–35
    [Google Scholar]
  38. Winocour E., Robbins E. 1970; Histone synthesis in polyoma- and SV40-infected cells. Virology 40:307–315
    [Google Scholar]
  39. Wohlheuter R. M., Marz R., Graff J. C., Plagemann P. G. W. 1976; The application of rapid kinetic techniques to the transport of thymidine and 3-O-methylglucose into mammalian cells in suspension culture. Journal of Cellular Physiology 89:605–612
    [Google Scholar]
  40. Zweerink H. J., Joklik W. J. 1970; Studies on the intracellular synthesis of reovirus-specified proteins. Virology 41501–518
    [Google Scholar]
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