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. 1980 Jun 11;8(11):2349–2363. doi: 10.1093/nar/8.11.2349

Virion DNA-independent RNA polymerase from Saccharomyces cerevisiae.

J D Welsh, M J Leibowitz, R B Wickner
PMCID: PMC324086  PMID: 7003533

Abstract

The "killer" plasmid and a larger double-stranded RNA plasmid of yeast exist in intracellular virion particles. Purification of these particles from a diploid killer strain of yeast (grown into stationary growth on ethanol) resulted in co-purification of a DNA-independent RNA polymerase activity. This activity incorporates and requires all four ribonucleoside triphosphates and will not act on deoxyribonucleoside triphosphates. The reaction requires magnesium, is inhibited by sulfhydryl-oxidizing reagents and high concentrations of monovalent cation, but is insensitive to DNase, alpha-amanitin, and actinomycin D. Pyrophosphate inhibits the reaction as does ethidium bromide. Exogenous nucleic acids have no effect on the reaction. The product is mostly single-stranded RNA, some of which is released from the enzymatically active virions.

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Selected References

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

  1. Adler J., Wood H. A., Bozarth R. F. Virus-like particles from killer, neutral, and sensitive strains of Saccharomyces cerevisiae. J Virol. 1976 Feb;17(2):472–476. doi: 10.1128/jvi.17.2.472-476.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bevan E. A., Herring A. J., Mitchell D. J. Preliminary characterization of two species of dsRNA in yeast and their relationship to the "killer" character. Nature. 1973 Sep 14;245(5420):81–86. doi: 10.1038/245081b0. [DOI] [PubMed] [Google Scholar]
  3. Black D. R., Knight C. A. Ribonucleic acid transcriptase acitvity in purified wound tumor virus. J Virol. 1970 Aug;6(2):194–198. doi: 10.1128/jvi.6.2.194-198.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Borsa J., Graham A. F. Reovirus: RNA polymerase activity in purified virions. Biochem Biophys Res Commun. 1968 Dec 30;33(6):895–901. doi: 10.1016/0006-291x(68)90396-3. [DOI] [PubMed] [Google Scholar]
  5. Buck K. W. Replication of double-stranded RNA in particles of Penicillium stoloniferum virus S. Nucleic Acids Res. 1975 Oct;2(10):1889–1902. doi: 10.1093/nar/2.10.1889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buck K. W. Semi-conservative replication of double-stranded RNA by a virion-associated RNA polymerase. Biochem Biophys Res Commun. 1978 Oct 16;84(3):639–645. doi: 10.1016/0006-291x(78)90753-2. [DOI] [PubMed] [Google Scholar]
  7. Cashel M., Lazzarini R. A., Kalbacher B. An improved method for thin-layer chromatography of nucleotide mixtures containing 32P-labelled orthophosphate. J Chromatogr. 1969 Mar 11;40(1):103–109. doi: 10.1016/s0021-9673(01)96624-5. [DOI] [PubMed] [Google Scholar]
  8. Chater K. F., Morgan D. H. Ribonucleic acid synthesis by isolated viruses of Penicillium stoloniferum. J Gen Virol. 1974 Aug;24(2):307–317. doi: 10.1099/0022-1317-24-2-307. [DOI] [PubMed] [Google Scholar]
  9. Cohen J. Ribonucleic acid polymerase activity associated with purified calf rotavirus. J Gen Virol. 1977 Sep;36(3):395–402. doi: 10.1099/0022-1317-36-3-395. [DOI] [PubMed] [Google Scholar]
  10. Cohn M. S., Tabor C. W., Tabor H., Wickner R. B. Spermidine or spermine requirement for killer double-stranded RNA plasmid replication in yeast. J Biol Chem. 1978 Aug 10;253(15):5225–5227. [PubMed] [Google Scholar]
  11. FRAENKEL-CONRAT H., SINGER B., TSUGITA A. Purification of viral RNA by means of bentonite. Virology. 1961 May;14:54–58. doi: 10.1016/0042-6822(61)90131-3. [DOI] [PubMed] [Google Scholar]
  12. Faust M., Millward S. In vitro methylation of nascent reovirus mRNA by a virion-associated methyl transferase. Nucleic Acids Res. 1974 Dec;1(12):1739–1752. doi: 10.1093/nar/1.12.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fink G. R., Styles C. A. Curing of a killer factor in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2846–2849. doi: 10.1073/pnas.69.10.2846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Furuichi Y. "Methylation-coupled" transcription by virus-associated transcriptase of cytoplasmic polyhedrosis virus containing double-stranded RNA. Nucleic Acids Res. 1974 Jun;1(6):809–822. doi: 10.1093/nar/1.6.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gomatos P. J., Kuechenthal I. Reovirus-specific enzyme(s) associated with subviral particles responds in vitro to polyribocytidylate to yield double-stranded polyribocytidylate-polyriboguanylate. J Virol. 1977 Jul;23(1):80–90. doi: 10.1128/jvi.23.1.80-90.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Groot G. S., Poyton R. O. Oxygen control of cytochrome c oxidase synthesis in isolated mitochondria from Saccharomyces cerevisiae. Nature. 1975 May 15;255(5505):238–240. doi: 10.1038/255238a0. [DOI] [PubMed] [Google Scholar]
  17. Groves W. E., Davis F. C., Jr, Sells B. H. Spectrophotometric determination of microgram quantities of protein without nucleic acid interference. Anal Biochem. 1968 Feb;22(2):195–210. doi: 10.1016/0003-2697(68)90307-2. [DOI] [PubMed] [Google Scholar]
  18. Hafe L. A., Keller E. B. The polyadenylate polymerases from yeast. J Biol Chem. 1975 Mar 10;250(5):1838–1846. [PubMed] [Google Scholar]
  19. Herring A. J., Bevan E. A. Virus-like particles associated with the double-stranded RNA species found in killer and sensitive strains of the yeast Saccharomyces cerevisiae. J Gen Virol. 1974 Mar;22(3):387–394. doi: 10.1099/0022-1317-22-3-387. [DOI] [PubMed] [Google Scholar]
  20. Holland M. J., Hager G. L., Rutter W. J. Characterization of purified poly(adenylic acid)-containing messenger ribonucleic acid from Saccharomyces cerevisiae. Biochemistry. 1977 Jan 11;16(1):8–16. doi: 10.1021/bi00620a002. [DOI] [PubMed] [Google Scholar]
  21. Holm C. A., Oliver S. G., Newman A. M., Holland L. E., McLaughlin C. S., Wagner E. K., Warner R. C. The molecular weight of yeast P1 double-stranded RNA. J Biol Chem. 1978 Nov 25;253(22):8332–8336. [PubMed] [Google Scholar]
  22. Hopper J. E., Bostian K. A., Rowe L. B., Tipper D. J. Translation of the L-species dsRNA genome of the killer-associated virus-like particles of Saccharomyces cerevisiae. J Biol Chem. 1977 Dec 25;252(24):9010–9017. [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. Lapierre H., Astier-Manifacier S., Cornuet P. Activité RNA polymérase associée aux préparations purifiées de virus du Penicillium stoloniferum. C R Acad Sci Hebd Seances Acad Sci D. 1971 Sep 13;273(11):992–994. [PubMed] [Google Scholar]
  25. Leibowitz M. J., Wickner R. B. Pet18: a chromosomal gene required for cell growth and for the maintenance of mitochondrial DNA and the killer plasmid of yeast. Mol Gen Genet. 1978 Oct 4;165(2):115–121. doi: 10.1007/BF00269899. [DOI] [PubMed] [Google Scholar]
  26. Lewandowski L. J., Kalmakoff J., Tanada Y. Characterization of a Ribonucleic Acid Polymerase Activity Associated with Purified Cytoplasmic Polyhedrosis Virus of the Silkworm Bombyx mori. J Virol. 1969 Dec;4(6):857–865. doi: 10.1128/jvi.4.6.857-865.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Martin S. A., Zweerink H. J. Isolation and characterization of two types of bluetongue virus particles. Virology. 1972 Nov;50(2):495–506. doi: 10.1016/0042-6822(72)90400-x. [DOI] [PubMed] [Google Scholar]
  28. Nash C. H., Douthart R. J., Ellis L. F., Van Frank R. M., Burnett J. P., Lemke P. A. On the mycophage of Penicillium chrysogenum. Can J Microbiol. 1973 Jan;19(1):97–103. doi: 10.1139/m73-014. [DOI] [PubMed] [Google Scholar]
  29. Oliver S. G., McCREADY S. J., Holm C., Sutherland P. A., McLaughlin C. S., Cox B. S. Biochemical and physiological studies of the yeast virus-like particle. J Bacteriol. 1977 Jun;130(3):1303–1309. doi: 10.1128/jb.130.3.1303-1309.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ratti G., Buck K. W. RNA polymerase activity in double-stranded ribonucleic acid virus particles from Aspergillus foetidus. Biochem Biophys Res Commun. 1975 Sep 16;66(2):706–711. doi: 10.1016/0006-291x(75)90567-7. [DOI] [PubMed] [Google Scholar]
  31. Reich E., Goldberg I. H. Actinomycin and nucleic acid function. Prog Nucleic Acid Res Mol Biol. 1964;3:183–234. doi: 10.1016/s0079-6603(08)60742-4. [DOI] [PubMed] [Google Scholar]
  32. Shatkin A. J., Sipe J. D. RNA polymerase activity in purified reoviruses. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1462–1469. doi: 10.1073/pnas.61.4.1462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Silverstein S. C., Christman J. K., Acs G. The reovirus replicative cycle. Annu Rev Biochem. 1976;45:375–408. doi: 10.1146/annurev.bi.45.070176.002111. [DOI] [PubMed] [Google Scholar]
  34. Somers J. M., Bevan E. A. The inheritance of the killer character in yeast. Genet Res. 1969 Feb;13(1):71–83. doi: 10.1017/s0016672300002743. [DOI] [PubMed] [Google Scholar]
  35. Stoltzfus C. M., Morgan M., Banerjee A. K., Shatkin A. J. Poly(A) polymerase activity in reovirus. J Virol. 1974 Jun;13(6):1338–1345. doi: 10.1128/jvi.13.6.1338-1345.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Van Etten J. L., Vidaver A. K., Koski R. K., Semancik J. S. RNA polymerase activity associated with bacteriophage phi 6. J Virol. 1973 Sep;12(3):464–471. doi: 10.1128/jvi.12.3.464-471.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Verwoerd D. W., Huismans H. Studies on the in vitro and the in vivo transcription of the bluetongue virus genome. Onderstepoort J Vet Res. 1972 Dec;39(4):185–191. [PubMed] [Google Scholar]
  38. Vodkin M., Katterman F., Fink G. R. Yeast killer mutants with altered double-stranded ribonucleic acid. J Bacteriol. 1974 Feb;117(2):681–686. doi: 10.1128/jb.117.2.681-686.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wehrli W., Knüsel F., Schmid K., Staehelin M. Interaction of rifamycin with bacterial RNA polymerase. Proc Natl Acad Sci U S A. 1968 Oct;61(2):667–673. doi: 10.1073/pnas.61.2.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wickner R. B. Chromosomal and nonchromosomal mutations affecting the "killer character" of Saccharomyces cerevisiae. Genetics. 1974 Mar;76(3):423–432. doi: 10.1093/genetics/76.3.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wickner R. B. Deletion of mitochondrial DNA bypassing a chromosomal gene needed for maintenance of the killer plasmid of yeast. Genetics. 1977 Nov;87(3):441–452. doi: 10.1093/genetics/87.3.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wickner R. B. Killer of Saccharomyces cerevisiae: a double-stranded ribonucleic acid plasmid. Bacteriol Rev. 1976 Sep;40(3):757–773. doi: 10.1128/br.40.3.757-773.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wickner R. B., Leibowitz M. J. Chromosomal genes essential for replication of a double-stranded RNA plasmid of Saccharomyces cerevisiae: the killer character of yeast. J Mol Biol. 1976 Aug 15;105(3):427–443. doi: 10.1016/0022-2836(76)90102-9. [DOI] [PubMed] [Google Scholar]
  44. Wickner R. B., Leibowitz M. J. Mak mutants of yeast: mapping and characterization. J Bacteriol. 1979 Oct;140(1):154–160. doi: 10.1128/jb.140.1.154-160.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wickner R. B., Leibowitz M. J. Two chromosomal genes required for killing expression in killer strains of Saccharomyces cerevisiae. Genetics. 1976 Mar 25;82(3):429–442. doi: 10.1093/genetics/82.3.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wickner R. B. The killer double-stranded RNA plasmids of yeast. Plasmid. 1979 Jul;2(3):303–322. doi: 10.1016/0147-619x(79)90015-5. [DOI] [PubMed] [Google Scholar]
  47. Wickner R. B. Twenty-six chromosomal genes needed to maintain the killer double-stranded RNA plasmid of Saccharomyces cerevisiae. Genetics. 1978 Mar;88(3):419–425. doi: 10.1093/genetics/88.3.419. [DOI] [PMC free article] [PubMed] [Google Scholar]

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