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. 1970 Jul;66(3):677–684. doi: 10.1073/pnas.66.3.677

Formation of a Vaccinia Virus Structural Polypeptide from a Higher Molecular Weight Precursor: Inhibition by Rifampicin

Ehud Katz 1, Bernard Moss 1
PMCID: PMC283104  PMID: 5269234

Abstract

A vaccinia virus core polypeptide, with a molecular weight of 76,000 and a relative deficiency in tryptophan, was shown by pulse-chase experiments to form from a precursor. The latter may be a rapidly labeled, 125,000-molecular weight, tryptophan-deficient, virus-induced polypeptide, which diminished in quantity during the chase period and was barely detectable after two to three hours.

Rifampicin completely prevented the formation of the core polypeptide without inhibiting the synthesis of the precursor. A rifampicin-resistant vaccinia mutant was used to demonstrate the specificity of this effect. The sequence of events after the removal of the drug suggested that cleavage of the precursor occurs during the formation of the virus core. Rifampicin appears to act by interrupting earlier maturational events which precede the formation of the core polypeptide.

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Selected References

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

  1. DAVIE E. W., RATNOFF O. D. WATERFALL SEQUENCE FOR INTRINSIC BLOOD CLOTTING. Science. 1964 Sep 18;145(3638):1310–1312. doi: 10.1126/science.145.3638.1310. [DOI] [PubMed] [Google Scholar]
  2. Dales S., Mosbach E. H. Vaccinia as a model for membrane biogenesis. Virology. 1968 Aug;35(4):564–583. doi: 10.1016/0042-6822(68)90286-9. [DOI] [PubMed] [Google Scholar]
  3. Dunker A. K., Rueckert R. R. Observations on molecular weight determinations on polyacrylamide gel. J Biol Chem. 1969 Sep 25;244(18):5074–5080. [PubMed] [Google Scholar]
  4. Easterbrook K. B. Controlled degradation of vaccinia virions in vitro: an electron microscopic study. J Ultrastruct Res. 1966 Mar;14(5):484–496. doi: 10.1016/s0022-5320(66)80077-1. [DOI] [PubMed] [Google Scholar]
  5. Eddleman H. L., Champe S. P. Components in T4-infected cells associated with phage assembly. Virology. 1966 Nov;30(3):471–481. doi: 10.1016/0042-6822(66)90123-1. [DOI] [PubMed] [Google Scholar]
  6. Fairbanks G., Jr, Levinthal C., Reeder R. H. Analysis of C14-labeled proteins by disc electrophoresis. Biochem Biophys Res Commun. 1965 Aug 16;20(4):393–399. doi: 10.1016/0006-291x(65)90589-9. [DOI] [PubMed] [Google Scholar]
  7. Gold P. H., Dales S. Localization of nucleotide phosphohydrolase activity within vaccinia. Proc Natl Acad Sci U S A. 1968 Jul;60(3):845–852. doi: 10.1073/pnas.60.3.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Heller E., Argaman M., Levy H., Goldblum N. Selective inhibition of vaccinia virus by the antibiotic rifampicin. Nature. 1969 Apr 19;222(5190):273–274. doi: 10.1038/222273a0. [DOI] [PubMed] [Google Scholar]
  9. Holland J. J., Kiehn E. D. Specific cleavage of viral proteins as steps in the synthesis and maturation of enteroviruses. Proc Natl Acad Sci U S A. 1968 Jul;60(3):1015–1022. doi: 10.1073/pnas.60.3.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jacobson M. F., Baltimore D. Morphogenesis of poliovirus. I. Association of the viral RNA with coat protein. J Mol Biol. 1968 Apr 28;33(2):369–378. doi: 10.1016/0022-2836(68)90195-2. [DOI] [PubMed] [Google Scholar]
  11. Jacobson M. F., Baltimore D. Polypeptide cleavages in the formation of poliovirus proteins. Proc Natl Acad Sci U S A. 1968 Sep;61(1):77–84. doi: 10.1073/pnas.61.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Joklik W. K. The poxviruses. Annu Rev Microbiol. 1968;22:359–390. doi: 10.1146/annurev.mi.22.100168.002043. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Kates J., Dahl R., Mielke M. Synthesis and intracellular localization of vaccinia virus deoxyribonucleic acid-dependent ribonucleic acid polymerase. J Virol. 1968 Sep;2(9):894–900. doi: 10.1128/jvi.2.9.894-900.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mcauslan B. R. Rifampicin inhibition of vaccinia replication. Biochem Biophys Res Commun. 1969 Oct 8;37(2):289–295. doi: 10.1016/0006-291x(69)90733-5. [DOI] [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., Rosenblum E. N. Vaccinia virus directed RNA and protein synthesis in the presence of rifampicin. Biochem Biophys Res Commun. 1969 Aug 22;36(5):858–865. doi: 10.1016/0006-291x(69)90688-3. [DOI] [PubMed] [Google Scholar]
  18. Moss B., Rosenblum E. N., Katz E., Grimley P. M. Rifampicin: a specific inhibitor of vaccinia virus assembly. Nature. 1969 Dec 27;224(5226):1280–1284. doi: 10.1038/2241280a0. [DOI] [PubMed] [Google Scholar]
  19. 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]
  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. Munyon W., Paoletti E., Ospina J., Grace J. T., Jr Nucleotide phosphohydrolase in purified vaccinia virus. J Virol. 1968 Mar;2(3):167–172. doi: 10.1128/jvi.2.3.167-172.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pitkanen A., McAuslan B., Hedgpeth J., Woodson B. Induction of poxvirus ribonucleic acid polymerases. J Virol. 1968 Dec;2(12):1363–1367. doi: 10.1128/jvi.2.12.1363-1367.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pogo B. G., Dales S. Two deoxyribonuclease activities within purified vaccinia virus. Proc Natl Acad Sci U S A. 1969 Jul;63(3):820–827. doi: 10.1073/pnas.63.3.820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  27. Steiner D. F., Oyer P. E. The biosynthesis of insulin and a probable precursor of insulin by a human islet cell adenoma. Proc Natl Acad Sci U S A. 1967 Feb;57(2):473–480. doi: 10.1073/pnas.57.2.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Subak-Sharpe J. H., Timbury M. C., Williams J. F. Rifampicin inhibits the growth of some mammalian viruses. Nature. 1969 Apr 26;222(5191):341–345. doi: 10.1038/222341a0. [DOI] [PubMed] [Google Scholar]
  29. Summers D. F., Maizel J. V., Jr Evidence for large precursor proteins in poliovirus synthesis. Proc Natl Acad Sci U S A. 1968 Mar;59(3):966–971. doi: 10.1073/pnas.59.3.966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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