Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1983 Feb 25;11(4):1077–1097. doi: 10.1093/nar/11.4.1077

Yeast killer dsRNA plasmids are transcribed in vivo to produce full and partial-length plus-stranded RNAs.

K A Bostian, V E Burn, S Jayachandran, D J Tipper
PMCID: PMC325777  PMID: 6338480

Abstract

In vivo transcripts of the L (4.5 kb) and M (1.9 kb) dsRNA plasmids were examined in type I killers of Saccharomyces cerevisiae. Transcripts for both plasmids include full-length (l,m) and partial-length (la,ma) single-stranded species. Both L-dsRNA transcripts (l,la) have in vitro mRNA activity for L-P1, previously shown to be identical to ScV-P1, the 88,000 dalton major capsid protein of the virus-like particles containing L- and M1-dsRNAs. 1, but not 1a, is bound to poly(U)-sepharose and may be polyadenylated. Other L-dsRNA gene products and their transcripts may exist. For M1-dsRNA, both species (m, ma) have in vitro mRNA activity for M1-P1, the 32,000 dalton pre-protoxin encoded by M1-dsRNA. Both m and ma are bound to poly(U)-Sepharose and ma is probably a 5' terminal fragment of m. A functional model for M1-dsRNA killer plasmid structure is presented.

Full text

PDF

Images in this article

1082Image
on p.1082
1085Image
on p.1085
1087Image
on p.1087
1088Image
on p.1088
1089Image
on p.1089
1091Image
on p.1091
1092Image
on p.1092

Selected References

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

  1. Bostian K. A., Hopper J. E., Rogers D. T., Tipper D. J. Translational analysis of the killer-associated virus-like particle dsRNA genome of S. cerevisiae: M dsRNA encodes toxin. Cell. 1980 Feb;19(2):403–414. doi: 10.1016/0092-8674(80)90514-0. [DOI] [PubMed] [Google Scholar]
  2. Bostian K. A., Lee R. C., Halvorson H. O. Preparative fractionation of nucleic acids by agarose gel electrophoresis. Anal Biochem. 1979 May;95(1):174–182. doi: 10.1016/0003-2697(79)90201-x. [DOI] [PubMed] [Google Scholar]
  3. Bostian K. A., Sturgeon J. A., Tipper D. J. Encapsidation of yeast killer double-stranded ribonucleic acids: dependence of M on L. J Bacteriol. 1980 Jul;143(1):463–470. doi: 10.1128/jb.143.1.463-470.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brennan V. E., Bobek L. A., Bruenn J. A. Yeast deRNA viral transcriptase pause products: identification of the transcript strand. Nucleic Acids Res. 1981 Oct 10;9(19):5049–5059. doi: 10.1093/nar/9.19.5049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bruenn J. A., Brennan V. E. Yeast viral double-stranded RNAs have heterogeneous 3' termini. Cell. 1980 Apr;19(4):923–933. doi: 10.1016/0092-8674(80)90084-7. [DOI] [PubMed] [Google Scholar]
  6. Bruenn J. A. Virus-like particles of yeast. Annu Rev Microbiol. 1980;34:49–68. doi: 10.1146/annurev.mi.34.100180.000405. [DOI] [PubMed] [Google Scholar]
  7. Bruenn J., Bobek L., Brennan V., Held W. Yeast viral RNA polymerase is a transcriptase. Nucleic Acids Res. 1980 Jul 11;8(13):2985–2997. doi: 10.1093/nar/8.13.2985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bussey H., Sacks W., Galley D., Saville D. Yeast killer plasmid mutations affecting toxin secretion and activity and toxin immunity function. Mol Cell Biol. 1982 Apr;2(4):346–354. doi: 10.1128/mcb.2.4.346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Darzynkiewicz E., Shatkin A. J. Assignment of reovirus mRNA ribosome binding sites to virion genome segments by nucleotide sequence analyses. Nucleic Acids Res. 1980 Jan 25;8(2):337–350. doi: 10.1093/nar/8.2.337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Field L. J., Bobek L. A., Brennan V. E., Reilly J. D., Bruenn J. A. There are at least two yeast viral double-stranded RNAs of the same size: an explanation for viral exclusion. Cell. 1982 Nov;31(1):193–200. doi: 10.1016/0092-8674(82)90419-6. [DOI] [PubMed] [Google Scholar]
  11. Franklin R. M. Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1504–1511. doi: 10.1073/pnas.55.6.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fried H. M., Fink G. R. Electron microscopic heteroduplex analysis of "killer" double-stranded RNA species from yeast. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4224–4228. doi: 10.1073/pnas.75.9.4224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Holland M. J., Holland J. P. Isolation and identification of yeast messenger ribonucleic acids coding for enolase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase. Biochemistry. 1978 Nov 14;17(23):4900–4907. doi: 10.1021/bi00616a007. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Iverson L. E., Rose J. K. Localized attenuation and discontinuous synthesis during vesicular stomatitis virus transcription. Cell. 1981 Feb;23(2):477–484. doi: 10.1016/0092-8674(81)90143-4. [DOI] [PubMed] [Google Scholar]
  17. Palatnik C. M., Storti R. V., Jacobson A. Fractionation and functional analysis of newly synthesized and decaying messenger RNAs from vegetative cells of Dictyostelium discoideum. J Mol Biol. 1979 Mar 5;128(3):371–395. doi: 10.1016/0022-2836(79)90093-7. [DOI] [PubMed] [Google Scholar]
  18. Palfree R. G., Bussey H. Yeast killer toxin: purification and characterisation of the protein toxin from Saccharomyces cerevisiae. Eur J Biochem. 1979 Feb 1;93(3):487–493. doi: 10.1111/j.1432-1033.1979.tb12847.x. [DOI] [PubMed] [Google Scholar]
  19. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  20. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Sogin S. J., Saunders C. A. Fluctuation in polyadenylate size and content in exponential- and stationary-phase cells of Saccharomyces cerevisiae. J Bacteriol. 1980 Oct;144(1):74–81. doi: 10.1128/jb.144.1.74-81.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sommer S. S., Wickner R. B. Co-curing of plasmids affecting killer double-stranded RNAs of Saccharomyces cerevisiae: [HOK], [NEX], and the abundance of L are related and further evidence that M1 requires L. J Bacteriol. 1982 May;150(2):545–551. doi: 10.1128/jb.150.2.545-551.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sommer S. S., Wickner R. B. Yeast L dsRNA consists of at least three distinct RNAs; evidence that the non-Mendelian genes [HOK], [NEX] and [EXL] are on one of these dsRNAs. Cell. 1982 Dec;31(2 Pt 1):429–441. doi: 10.1016/0092-8674(82)90136-2. [DOI] [PubMed] [Google Scholar]
  25. Thiele D. J., Wang R. W., Leibowitz M. J. Separation and sequence of the 3' termini of M double-stranded RNA from killer yeast. Nucleic Acids Res. 1982 Mar 11;10(5):1661–1678. doi: 10.1093/nar/10.5.1661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Toh-E A., Wickner R. B. "Superkiller" mutations suppress chromosomal mutations affecting double-stranded RNA killer plasmid replication in saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jan;77(1):527–530. doi: 10.1073/pnas.77.1.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Welsh D., Leibowitz M. J. Transcription of killer virion double-stranded RNA in vitro. Nucleic Acids Res. 1980 Jun 11;8(11):2365–2375. doi: 10.1093/nar/8.11.2365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wickner R. B. Plasmids controlled exclusion of the K2 killer double-stranded RNA plasmid of yeast. Cell. 1980 Aug;21(1):217–226. doi: 10.1016/0092-8674(80)90129-4. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. de la Peña P., Barros F., Gascón S., Lazo P. S., Ramos S. Effect of yeast killer toxin on sensitive cells of Saccharomyces cerevisiae. J Biol Chem. 1981 Oct 25;256(20):10420–10425. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES