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. 1974 Sep;14(3):704–708. doi: 10.1128/jvi.14.3.704-708.1974

Evidence for the Presence of RNA in the Purified Virions of Vaccinia Virus

G Roening 1, J A Holowczak 1
PMCID: PMC355566  PMID: 4851942

Abstract

Vaccinia virus, strain WR, was propagated in HeLa cells, L mouse fibroblats, or primary chicken embryo fibroblasts in the presence of [5- 3H]uridine. Carefully purified virions were found to contain significant amounts of labeled trichloroacetic acid-precipitable material which was rendered acid soluble when digested with pancreatic RNase or hydrolyzed in alkali. Controlled degradation of virions with Nonidet P-40 and 2-mercaptoethanol demonstrated that 65 to 80% of the [5- 3H]uridine-labeled molecules resided in the viral core. When the total nucleic acids were extracted from viral cores prepared from virions propagated in HeLa cells, 30 to 50% of the total incorporated [5- 3H]uridine was found in RNA; in L mouse fibroblasts, 40 to 50%; in primary chicken embryo fibroblasts, 50 to 60%. The RNA molecules do not appear to be covalently linked to the viral DNA genome but sediment in sodium dodecyl sulfate-sucrose gradients as 8 to 10S species relative to ribosomal RNA.

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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. Becker Y., Sarov I. Electron microscopy of Vaccinia virus DNA. J Mol Biol. 1968 Jun 28;34(3):655–660. doi: 10.1016/0022-2836(68)90187-3. [DOI] [PubMed] [Google Scholar]
  3. Brown M., Dorson J. W., Bollum F. J. Terminal riboadenylate transferase: a poly A polymerase in purified vaccinia virus. J Virol. 1973 Aug;12(2):203–208. doi: 10.1128/jvi.12.2.203-208.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burlingham B. T., Doerfler W. Three size-classes of intracellular adenovirus deoxyribonucleic acid. J Virol. 1971 Jun;7(6):707–719. doi: 10.1128/jvi.7.6.707-719.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Fox R. M., Mendelsohn J., Barbosa E., Goulian M. RNA in nascent DNA from cultured human lymphocytes. Nat New Biol. 1973 Oct 24;245(147):234–237. doi: 10.1038/newbio245234a0. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Holowczak J. A. Uncoating of poxviruses. I. Detection and characterization of subviral particles in the uncoating process. Virology. 1972 Oct;50(1):216–232. doi: 10.1016/0042-6822(72)90362-5. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. 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]
  13. Kates J., Beeson J. Ribonucleic acid synthesis in vaccinia virus. I. The mechanism of synthesis and release of RNA in vaccinia cores. J Mol Biol. 1970 May 28;50(1):1–18. doi: 10.1016/0022-2836(70)90100-2. [DOI] [PubMed] [Google Scholar]
  14. Kleiman J., Moss B. Protein kinase activity from vaccinia virions: solubilization and separation into heat-labile and heat-stable components. J Virol. 1973 Oct;12(4):684–689. doi: 10.1128/jvi.12.4.684-689.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lark K. G. Evidence for the direct involvement of RNA in the initiation of DNA replication in Escherichia coli 15T. J Mol Biol. 1972 Feb 28;64(1):47–60. doi: 10.1016/0022-2836(72)90320-8. [DOI] [PubMed] [Google Scholar]
  16. Lucas J. J., Ginsberg H. S. Transcription and transport of virus-specific ribonucleic acids in African green monkey kidney cells abortively infected with type 2 adenovirus. J Virol. 1972 Dec;10(6):1109–1117. doi: 10.1128/jvi.10.6.1109-1117.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. 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]
  20. Oki T., Fujiwara Y., Heidelberger C. Utilization of host-cell DNA by vaccinia virus replicating in HeLa cells irradiated intranuclearly with tritium. J Gen Virol. 1971 Dec;13(3):401–413. doi: 10.1099/0022-1317-13-3-401. [DOI] [PubMed] [Google Scholar]
  21. PLANTEROSE D. N., NISHIMURA C., SALZMAN N. P. The purification of vaccinia virus from cell cultures. Virology. 1962 Oct;18:294–301. doi: 10.1016/0042-6822(62)90016-8. [DOI] [PubMed] [Google Scholar]
  22. Paoletti E., Moss B. Protein kinase and specific phosphate acceptor proteins associated with vaccinia virus cores. J Virol. 1972 Sep;10(3):417–424. doi: 10.1128/jvi.10.3.417-424.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Parkhurst J. R., Peterson A. R., Heidelberger C. Breakdown of HeLa cell DNA mediated by vaccinia virus. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3200–3204. doi: 10.1073/pnas.70.11.3200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pogo B. G., Dales S. Regulation of the synthesis of nucleotide phosphohydrolase and neutral deoxyribonuclease: two activities present within purified vaccina virus. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1297–1303. doi: 10.1073/pnas.63.4.1297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Sarov I., Becker Y. Studies on vaccinia virus DNA. Virology. 1967 Nov;33(3):369–375. doi: 10.1016/0042-6822(67)90112-2. [DOI] [PubMed] [Google Scholar]
  27. Speyer J. F., Chao J., Chao L. Ribonucleotides covalently linked to deoxyribonucleic acid in T4 bacteriophage. J Virol. 1972 Nov;10(5):902–908. doi: 10.1128/jvi.10.5.902-908.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sugino A., Hirose S., Okazaki R. RNA-linked nascent DNA fragments in Escherichia coli. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1863–1867. doi: 10.1073/pnas.69.7.1863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Waqar M. A., Huberman J. A. Evidence for the attachment of RNA to pulse-labeled DNA in the slime mold, Physarum polycephalum. Biochem Biophys Res Commun. 1973 Mar 5;51(1):174–180. doi: 10.1016/0006-291x(73)90524-x. [DOI] [PubMed] [Google Scholar]
  30. ZWARTOUW H. T. THE CHEMICAL COMPOSITION OF VACCINIA VIRUS. J Gen Microbiol. 1964 Jan;34:115–123. doi: 10.1099/00221287-34-1-115. [DOI] [PubMed] [Google Scholar]
  31. Zimmerman S. B., Sandeen D. The ribonuclease activity of crystallized pancreatic deoxyribonuclease. Anal Biochem. 1966 Feb;14(2):269–277. doi: 10.1016/0003-2697(66)90137-0. [DOI] [PubMed] [Google Scholar]

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