Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1970 Feb;5(2):99–108. doi: 10.1128/jvi.5.2.99-108.1970

Irreversible Effects of Cycloheximide During the Early Period of Vaccinia Virus Replication

Bernard Moss 1, Ronald Filler 1
PMCID: PMC375975  PMID: 5444010

Abstract

The presence of cycloheximide, an inhibitor of protein synthesis, during the period 30 to 60 min after vaccinia infection produced an irreversible block in virus replication. In contrast (i) cycloheximide given at earlier or later times, even for prolonged periods, did not prevent continuation of the infectious cycle after removal of the drug, and (ii) treatment with cycloheximide during the first 2 hr did not prevent virus growth when the early stages of replication proceeded more slowly due to infection with a low multiplicity of virus. These findings were interpreted as an indication that protein synthesis is required at a critical time in the virus growth cycle. Under the conditions in which brief cycloheximide treatment prevented virus growth, ribonucleic acid (RNA) synthesis continued at an undiminished rate for at least 2 hr after removal of the drug. Although this RNA appeared identical by polyacrylamide gel electrophoresis to “early” viral messenger RNA, it was not found associated with ribosomes or polyribosomes. Failure to observe viral protein synthesis was consistent with the latter finding. It appeared unlikely that the translational block resulted from inadequate removal of cycloheximide, since the effects of the drug were shown to be reversible at earlier or later times in infection or even at the same time when a lower multiplicity of virus was used. Interference with the normal synthesis of specific viral protein factors required for translation was postulated to explain the results.

Full text

PDF
99

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BECKER Y., JOKLIK W. K. MESSENGER RNA IN CELLS INFECTED WITH VACCINIA VIRUS. Proc Natl Acad Sci U S A. 1964 Apr;51:577–585. doi: 10.1073/pnas.51.4.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bishop D. H., Claybrook J. R., Spiegelman S. Electrophoretic separation of viral nucleic acids on polyacrylamide gels. J Mol Biol. 1967 Jun 28;26(3):373–387. doi: 10.1016/0022-2836(67)90310-5. [DOI] [PubMed] [Google Scholar]
  3. Chrambach A., Reisfeld R. A., Wyckoff M., Zaccari J. A procedure for rapid and sensitive staining of protein fractionated by polyacrylamide gel electrophoresis. Anal Biochem. 1967 Jul;20(1):150–154. doi: 10.1016/0003-2697(67)90272-2. [DOI] [PubMed] [Google Scholar]
  4. DALES S. The uptake and development of vaccinia virus in strain L cells followed with labeled viral deoxyribonucleic acid. J Cell Biol. 1963 Jul;18:51–72. doi: 10.1083/jcb.18.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dales S. Effects of streptovitacin A on the initial events in the replication of vaccinia and reovirus. Proc Natl Acad Sci U S A. 1965 Aug;54(2):462–468. doi: 10.1073/pnas.54.2.462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  7. JOKLIK W. K., BECKER Y. THE REPLICATION AND COATING OF VACCINIA DNA. J Mol Biol. 1964 Dec;10:452–474. doi: 10.1016/s0022-2836(64)80066-8. [DOI] [PubMed] [Google Scholar]
  8. JOKLIK W. K. THE INTRACELLULAR UNCOATING OF POXVIRUS DNA. I. THE FATE OF RADIOACTIVELY-LABELED RABBITPOX VIRUS. J Mol Biol. 1964 Feb;8:263–276. doi: 10.1016/s0022-2836(64)80136-4. [DOI] [PubMed] [Google Scholar]
  9. JOKLIK W. K. The preparation and characteristics of highly purified radioactively labelled poxvirus. Biochim Biophys Acta. 1962 Aug 20;61:290–301. doi: 10.1016/0926-6550(62)90091-9. [DOI] [PubMed] [Google Scholar]
  10. Kates J. R., McAuslan B. R. Messenger RNA synthesis by a "coated" viral genome. Proc Natl Acad Sci U S A. 1967 Feb;57(2):314–320. doi: 10.1073/pnas.57.2.314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kates J. R., McAuslan B. R. Poxvirus DNA-dependent RNA polymerase. Proc Natl Acad Sci U S A. 1967 Jul;58(1):134–141. doi: 10.1073/pnas.58.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Katz E., Moss B. Synthesis of vaccinia viral proteins in cytoplasmic extracts. I. Incorporation of radioactively labeled amino acids into polypeptides. J Virol. 1969 Oct;4(4):416–422. doi: 10.1128/jvi.4.4.416-422.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Loening U. E. The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochem J. 1967 Jan;102(1):251–257. doi: 10.1042/bj1020251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Moss B., Ingram V. M. The repression and induction by thyroxin of hemoglobin synthesis during amphibian metamorphosis. Proc Natl Acad Sci U S A. 1965 Sep;54(3):967–974. doi: 10.1073/pnas.54.3.967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moss B. Inhibition of HeLa cell protein synthesis by the vaccinia virion. J Virol. 1968 Oct;2(10):1028–1037. doi: 10.1128/jvi.2.10.1028-1037.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Moss B., Katz E. Synthesis of vaccinia viral proteins in cytoplasmic extracts. II. Identification of early and late viral proteins. J Virol. 1969 Nov;4(5):596–602. doi: 10.1128/jvi.4.5.596-602.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moss B., Salzman N. P. Sequential protein synthesis following vaccinia virus infection. J Virol. 1968 Oct;2(10):1016–1027. doi: 10.1128/jvi.2.10.1016-1027.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Munyon W. H., Kit S. Induction of cytoplasmic ribonucleic acid (RNA) synthesis in vaccinia-infected LM cells during inhibition of protein synthesis. Virology. 1966 Jun;29(2):303–309. doi: 10.1016/0042-6822(66)90037-7. [DOI] [PubMed] [Google Scholar]
  20. Munyon W., Paoletti E., Grace J. T., Jr RNA polymerase activity in purified infectious vaccinia virus. Proc Natl Acad Sci U S A. 1967 Dec;58(6):2280–2287. doi: 10.1073/pnas.58.6.2280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Noll H. Chain initiation and control of protein synthesis. Science. 1966 Mar 11;151(3715):1241–1245. doi: 10.1126/science.151.3715.1241. [DOI] [PubMed] [Google Scholar]
  22. Oda K. I., Joklik W. K. Hybridization and sedimentation studies on "early" and "late" vaccinia messenger RNA. J Mol Biol. 1967 Aug 14;27(3):395–419. doi: 10.1016/0022-2836(67)90047-2. [DOI] [PubMed] [Google Scholar]
  23. SALZMAN N. P., SHATKIN A. J., SEBRING E. D. THE SYNTHESIS OF A DNA-LIKE RNA IN THE CYTOPLASM OF HELA CELLS INFECTED WITH VACCINIA VIRUS. J Mol Biol. 1964 Mar;8:405–416. doi: 10.1016/s0022-2836(64)80204-7. [DOI] [PubMed] [Google Scholar]
  24. SALZMAN N. P. The rate of formation of vaccinia deoxyribonucleic acid and vaccinia virus. Virology. 1960 Jan;10:150–152. doi: 10.1016/0042-6822(60)90015-5. [DOI] [PubMed] [Google Scholar]
  25. SHATKIN A. J. ACTINOMYCIN D AND VACCINIA VIRUS INFECTION OF HELA CELLS. Nature. 1963 Jul 27;199:357–358. doi: 10.1038/199357a0. [DOI] [PubMed] [Google Scholar]
  26. Salzman N. P., Sebring E. D. Sequential formation of vaccinia virus proteins and viral deoxyribonucleic acid replication. J Virol. 1967 Feb;1(1):16–23. doi: 10.1128/jvi.1.1.16-23.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. WARNER J. R., KNOPF P. M., RICH A. A multiple ribosomal structure in protein synthesis. Proc Natl Acad Sci U S A. 1963 Jan 15;49:122–129. doi: 10.1073/pnas.49.1.122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Woodson B. Vaccinia mRNA synthesis under conditions which prevent uncoating. Biochem Biophys Res Commun. 1967 Apr 20;27(2):169–175. doi: 10.1016/s0006-291x(67)80057-3. [DOI] [PubMed] [Google Scholar]
  29. Young R. W., Fulhorst H. W. Recovery of S35 radioactivity from protein-bearing polyacrylamide gel. Anal Biochem. 1965 May;11(2):389–391. doi: 10.1016/0003-2697(65)90030-8. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES