Abstract
The double-stranded RNA (dsRNA) viruses of Saccharomyces cerevisiae consist of 4.5-kilobase-pair (kb) L species and 1.7- to 2.1-kb M species, both found in cytoplasmic viruslike particles (VLPs). The L species encode their own capsid protein, and one (LA) has been shown to encode a putative capsid-polymerase fusion protein (cap-pol) that presumably provides VLPs with their transcriptase and replicase functions. The M1 and M2 dsRNAs encode the K1 and K2 toxins and specific immunity mechanisms. Maintenance of M1 and M2 is dependent on the presence of LA, which provides capsid and cap-pol for M dsRNA maintenance. Although a number of different S. cerevisiae killers have been described, only K1 and K2 have been studied in any detail. Their secreted polypeptide toxins disrupt cytoplasmic membrane functions in sensitive yeast cells. K28, named for the wine S. cerevisiae strain 28, appears to be unique; its toxin is unusually stable and disrupts DNA synthesis in sensitive cells. We have now demonstrated that 4.5-kb L28 and 2.1-kb M28 dsRNAs can be isolated from strain 28 in typical VLPs, that these VLPs are sufficient to confer K28 toxin and immunity phenotypes on transfected spheroplasts, and that the immunity of the transfectants is distinct from that of either M1 or M2. In vitro transcripts from the M28 VLPs show no cross-hybridization to denatured M1 or M2 dsRNAs, while L28 is an LA species competent for maintenance of M1. K28, encoded by M28, is thus the third unique killer system in S. cerevisiae to be clearly defined. It is now amenable to genetic analysis in standard laboratory strains.
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- Ball S. G., Tirtiaux C., Wickner R. B. Genetic Control of L-a and L-(Bc) Dsrna Copy Number in Killer Systems of SACCHAROMYCES CEREVISIAE. Genetics. 1984 Jun;107(2):199–217. doi: 10.1093/genetics/107.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bostian K. A., Elliott Q., Bussey H., Burn V., Smith A., Tipper D. J. Sequence of the preprotoxin dsRNA gene of type I killer yeast: multiple processing events produce a two-component toxin. Cell. 1984 Mar;36(3):741–751. doi: 10.1016/0092-8674(84)90354-4. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
- Diamond M. E., Dowhanick J. J., Nemeroff M. E., Pietras D. F., Tu C. L., Bruenn J. A. Overlapping genes in a yeast double-stranded RNA virus. J Virol. 1989 Sep;63(9):3983–3990. doi: 10.1128/jvi.63.9.3983-3990.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- El-Sherbeini M., Tipper D. J., Mitchell D. J., Bostian K. A. Virus-like particle capsid proteins encoded by different L double-stranded RNAs of Saccharomyces cerevisiae: their roles in maintenance of M double-stranded killer plasmids. Mol Cell Biol. 1984 Dec;4(12):2818–2827. doi: 10.1128/mcb.4.12.2818. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Esteban R., Wickner R. B. Three different M1 RNA-containing viruslike particle types in Saccharomyces cerevisiae: in vitro M1 double-stranded RNA synthesis. Mol Cell Biol. 1986 May;6(5):1552–1561. doi: 10.1128/mcb.6.5.1552. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Hanes S. D., Burn V. E., Sturley S. L., Tipper D. J., Bostian K. A. Expression of a cDNA derived from the yeast killer preprotoxin gene: implications for processing and immunity. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1675–1679. doi: 10.1073/pnas.83.6.1675. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hannig E. M., Leibowitz M. J. Structure and expression of the M2 genomic segment of a type 2 killer virus of yeast. Nucleic Acids Res. 1985 Jun 25;13(12):4379–4400. doi: 10.1093/nar/13.12.4379. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hill J. E., Myers A. M., Koerner T. J., Tzagoloff A. Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast. 1986 Sep;2(3):163–167. doi: 10.1002/yea.320020304. [DOI] [PubMed] [Google Scholar]
- 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]
- Icho T., Wickner R. B. The double-stranded RNA genome of yeast virus L-A encodes its own putative RNA polymerase by fusing two open reading frames. J Biol Chem. 1989 Apr 25;264(12):6716–6723. [PubMed] [Google Scholar]
- Lolle S., Skipper N., Bussey H., Thomas D. Y. The expression of cDNA clones of yeast M1 double-stranded RNA in yeast confers both killer and immunity phenotypes. EMBO J. 1984 Jun;3(6):1383–1387. doi: 10.1002/j.1460-2075.1984.tb01981.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pfeiffer P., Radler F. Comparison of the killer toxin of several yeasts and the purification of a toxin of type K2. Arch Microbiol. 1984 Apr;137(4):357–361. doi: 10.1007/BF00410734. [DOI] [PubMed] [Google Scholar]
- Ridley S. P., Sommer S. S., Wickner R. B. Superkiller mutations in Saccharomyces cerevisiae suppress exclusion of M2 double-stranded RNA by L-A-HN and confer cold sensitivity in the presence of M and L-A-HN. Mol Cell Biol. 1984 Apr;4(4):761–770. doi: 10.1128/mcb.4.4.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmitt M., Radler F. Mannoprotein of the yeast cell wall as primary receptor for the killer toxin of Saccharomyces cerevisiae strain 28. J Gen Microbiol. 1987 Dec;133(12):3347–3354. doi: 10.1099/00221287-133-12-3347. [DOI] [PubMed] [Google Scholar]
- Skipper N. Synthesis of a double-stranded cDNA transcript of the killer toxin-coding region of the yeast M1 double-stranded RNA. Biochem Biophys Res Commun. 1983 Jul 29;114(2):518–525. doi: 10.1016/0006-291x(83)90811-2. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Thiele D. J., Hannig E. M., Leibowitz M. J. Multiple L double-stranded RNA species of Saccharomyces cerevisiae: evidence for separate encapsidation. Mol Cell Biol. 1984 Jan;4(1):92–100. doi: 10.1128/mcb.4.1.92. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thiele D. J., Leibowitz M. J. Structural and functional analysis of separated strands of killer double-stranded RNA of yeast. Nucleic Acids Res. 1982 Nov 11;10(21):6903–6918. doi: 10.1093/nar/10.21.6903. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tipper D. J., Bostian K. A. Double-stranded ribonucleic acid killer systems in yeasts. Microbiol Rev. 1984 Jun;48(2):125–156. doi: 10.1128/mr.48.2.125-156.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Welsh J. D., Leibowitz M. J. Localization of genes for the double-stranded RNA killer virus of yeast. Proc Natl Acad Sci U S A. 1982 Feb;79(3):786–789. doi: 10.1073/pnas.79.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Welsh J. D., Leibowitz M. J., Wickner R. B. Virion DNA-independent RNA polymerase from Saccharomyces cerevisiae. Nucleic Acids Res. 1980 Jun 11;8(11):2349–2363. doi: 10.1093/nar/8.11.2349. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wickner R. B. "Killer character" of Saccharomyces cerevisiae: curing by growth at elevated temperature. J Bacteriol. 1974 Mar;117(3):1356–1357. doi: 10.1128/jb.117.3.1356-1357.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wickner R. B. Double-stranded RNA replication in yeast: the killer system. Annu Rev Biochem. 1986;55:373–395. doi: 10.1146/annurev.bi.55.070186.002105. [DOI] [PubMed] [Google Scholar]
- el-Sherbeini M., Bostian K. A. Viruses in fungi: infection of yeast with the K1 and K2 killer viruses. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4293–4297. doi: 10.1073/pnas.84.12.4293. [DOI] [PMC free article] [PubMed] [Google Scholar]