Sequences at the 3′ ends of yeast viral dsRNAs: proposed transcriptase and replicase initiation sites

Brennan, Victoria E.; Field, Loren J.; Cizdziel, Paul E.; Bruenn, Jeremy A.

doi:10.1093/nar/9.16.4007

Cited by 38 publications

(39 citation statements)

References 37 publications

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“…One possibility for this step is that some of the plus strands that were extruded are subsequently VOL. 6,1986 on May 10, 2018 by guest http://mcb.asm.org/ Downloaded from encapsidated by newly produced coat proteins. A second possibility is that encapsidation and plus ssRNA synthesis are coupled.…”

Section: Discussionmentioning

confidence: 99%

“…6,1986 on May 10, 2018 by guest http://mcb.asm.org/ Downloaded from denatured by heating at 83°C for 3 min in the presence of 90% formamide (final volume, 10 ,ul); then the same volume of 32P-labeled RNA polymerase reaction products, dissolved in 8 x SSC, was added. After another 1-min incubation at 83°C, annealing was begun at 46°C.…”

Section: Methodsmentioning

confidence: 99%

“…The phenolextracted RNA product is mainly single stranded as judged VOL. 6,1986 on May 10, 2018 by guest http://mcb.asm.org/ Downloaded from by its sensitivity to pancreatic RNase in the presence of high salt concentrations (Table 3). When pancreatic RNase was added to the reaction mixture after the RNA polymerase reaction was halted by adding 10 mM EDTA, the product of the M1-H VLPs was completely degraded, implying that it had been extruded from the particles (Fig.…”

Section: Methodsmentioning

confidence: 99%

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Three different M1 RNA-containing viruslike particle types in Saccharomyces cerevisiae: in vitro M1 double-stranded RNA synthesis.

Esteban¹,

Wickner²

1986

Mol. Cell. Biol.

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Killer strains of Saccharomyces cerevisiae bear at least two different double-stranded RNAs (dsRNAs) encapsidated in 39-nm viruslike particles (VLPs) of which the major coat protein is coded by the larger RNA (L-A dsRNA). The smaller dsRNA (M1 or M2) encodes an extracellular protein toxin (Kl or K2 toxin). Based on their densities on CsCl gradients, L-A-and Ml-containing particles can be separated. Using this method, we detected a new type of Ml dsRNA-containing VLP (Ml-H VLP, for heavy) that has a higher density than those previously reported (Ml-L VLP, for light). Ml-H and Ml-L VLPs are present together in the same strains and in all those we tested. Ml-H, Ml-L, and L-A VLPs all have the same types of proteins in the same approximate proportions, but whereas L-A VLPs and Ml-L VLPs have one dsRNA molecule per particle, MI-H VLPs contain two Ml dsRNA molecules per particle. Their RNA polymerase produces mainly plus single strands that are all extruded in the case of M1-H particles but are partially retained inside the Ml-L particles to be used later for dsRNA synthesis. We show that MI-H VLPs are formed in vitro from the M1-L VLPs. We also show that the peak of Ml dsRNA synthesis is in fractions lighter than Ml-L VLPs, presumably those carrying only a single plus M1 strand. We suggest that VLPs carrying two M1 dsRNAs (each 1.8 kilobases) can exist because the particle is designed to carry one L-A dsRNA (4.5 kilobases).

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Three different M1 RNA-containing viruslike particle types in Saccharomyces cerevisiae: in vitro M1 double-stranded RNA synthesis.

Esteban¹,

Wickner²

1986

Mol. Cell. Biol.

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“…They are differentiated by which of the three genes, [HOK], [NEX], and [EXL], they carry (Table 1) (17,25,31). L-A-H (the L2 of Brennan et al [4]) carries only [HOK] (R. B. Wickner, unpublished data). Wild-type K1 killer strains all carry L-A-HN (17,27), whereas wild-type K2 killers carry either L-A-HN or L-A-H (4,17,25;Wickner,unpublished data).…”

mentioning

confidence: 99%

“…ItsS was mapped near the centromere of chromosome XVI (30), and we show here that it is the same as mak6. Mutations in 11 other MAK genes (mak3, 4,7,8,10,11,12,16,18,19, and 27) do not cause cold sensitivity for growth (R. B. Wickner, unpublished data), and among the low temperature-sensitive mutants of Singh and Manney, only ltsS (mak6) shows the mak-phenotype (30). The ski mutants were first discovered based on their increased production of the K1 toxin (the superkiller phenotype) (18).…”

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confidence: 99%

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.

Ridley¹,

Sommer²,

Wickner³

1984

Mol. Cell. Biol.

145

158

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In an mktl host, L-A-HN double-stranded RNA excludes M2 double-stranded RNA at 30°C but not at 20°C. Recessive mutations suppressing the exclusion of M2 by L-A-HN in an mktl host include six ski (superkiller) genes, three of which (ski6, ski7 and ski8) are new genes. The dominant mutations in one gene (MKS50) and recessive mutations in at least two genes (mksl and mks2) suppress M2 exclusion by L-A-HN but do not show other characteristics of ski mutations and thus define a new class of killer-related chromosomal genes. Mutations in ski2, ski3, ski4, ski6, ski7, and ski8 result in increased M copy number at 30°C and prevent the cells from growing at 8°C. Elimination of M double-stranded RNA from a coldsensitive ski-strain results in the loss of cold sensitivity. ski-[KIL-sd1] strains lack L-A-HN, carry L-A-E, and have a lower M1 copy number than do ski- [KIL-kl] strains and are only slightly cold sensitive. The LTS5 (=MAK6) product is required both for low temperature growth and for M1 maintenance or replication. We propose that the elevated levels of M in ski-strains divert the host LTS5 product away from the host and to the M replication process. We also suggest that the essential role of L-A in M replication is protection of M double-stranded RNA from the negative influence of SKI' products.The killer system of Saccharomyces cerevisiae is unique among eucaryotic virus and plasmid systems in the detail in which interactions of viral and host components have been explored and in the large number of host components shown to be involved. This probably reflects the ease with which these aspects can be investigated in S. cerevisiae rather than any difference in the degree to which viral and host functions are intertwined. Numerous examples are known of host restriction of viral growth in mammalian systems, but analysis of the host genes involved is generally not practical. We show here that overproduction of a S. cerevisiae virus, due to a host mutation, results in host pathology, apparently due to over-utilization by the virus of a specific, essential host component.

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ScV “Killer” Viruses in Yeast

Mitchell

Bevan

1983

Springer Series in Molecular Biology

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Sequences at the 3′ ends of yeast viral dsRNAs: proposed transcriptase and replicase initiation sites

Cited by 38 publications

References 37 publications

Three different M1 RNA-containing viruslike particle types in Saccharomyces cerevisiae: in vitro M1 double-stranded RNA synthesis.

Three different M1 RNA-containing viruslike particle types in Saccharomyces cerevisiae: in vitro M1 double-stranded RNA synthesis.

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.

ScV “Killer” Viruses in Yeast

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