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. 2023 Sep 30;13(10):jkad167.
doi: 10.1093/g3journal/jkad167.

RNA viruses, M satellites, chromosomal killer genes, and killer/nonkiller phenotypes in the 100-genomes S. cerevisiae strains

Sriram Vijayraghavan  1 , Stanislav G Kozmin  1 , Pooja K Strope  1 , Daniel A Skelly  2 , Paul M Magwene  2 , Fred S Dietrich  1 , John H McCusker  1
Affiliations

RNA viruses, M satellites, chromosomal killer genes, and killer/nonkiller phenotypes in the 100-genomes S. cerevisiae strains

Sriram Vijayraghavan et al. G3 (Bethesda). .

Abstract

We characterized previously identified RNA viruses (L-A, L-BC, 20S, and 23S), L-A-dependent M satellites (M1, M2, M28, and Mlus), and M satellite-dependent killer phenotypes in the Saccharomyces cerevisiae 100-genomes genetic resource population. L-BC was present in all strains, albeit in 2 distinct levels, L-BChi and L-BClo; the L-BC level is associated with the L-BC genotype. L-BChi, L-A, 20S, 23S, M1, M2, and Mlus (M28 was absent) were in fewer strains than the similarly inherited 2µ plasmid. Novel L-A-dependent phenotypes were identified. Ten M+ strains exhibited M satellite-dependent killing (K+) of at least 1 of the naturally M0 and cured M0 derivatives of the 100-genomes strains; in these M0 strains, sensitivities to K1+, K2+, and K28+ strains varied. Finally, to complement our M satellite-encoded killer toxin analysis, we assembled the chromosomal KHS1 and KHR1 killer genes and used naturally M0 and cured M0 derivatives of the 100-genomes strains to assess and characterize the chromosomal killer phenotypes.

Keywords: Saccharomyces cerevisiae; 100-genomes strains; RNA virus; associations; killer; phenotypes.

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Conflict of interest statement

Conflicts of interest The authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Representative gels (top) of strains carrying different species of the dsRNA totiviruses (L-A and L-BC) and satellites (M1, M2, and Mlus). Shown are isogenic strain pairs of native and L-A–cured strains. (Bottom) Corresponding PCRs from cDNAs of RNA derived from the indicated strains, probing for presence/absence of L-A (A), L-BC (B), or M (M1, M2, and Mlus). All samples were treated with DNase prior to gel and PCR analysis. Primers for PCR analysis LA-F4 + LA-R5 (L-A), LBC-F2 + LBC-R2 (L-BC), M1-F + M1-R (M1), M2-F + M2-R (M2), and Mlus-F2 + Mlus-R2 (Mlus).
Fig. 2.
Fig. 2.
Killer activity in M-containing strains. Isogenic strain pairs of native and L-A–cured strains were tested on lawns of M0 strains. Representative strains displaying a) K1, b) K2, c) Klus, and d) K28 killer activity are shown. For Klus, only 1 strain background (YJM428) showed sensitivity to the Mlus toxin. For K28, a corresponding M0 strain was not tested.
Fig. 3.
Fig. 3.
Phenotypic analysis of a) YJM1133 and b) YJM1419 L-A+ M+ parental as well as isogenic L-A+ M0 and L-A0 M0 derivatives. Ten-fold dilutions of isogenic strains differing only in the presence/absence of L-A and/or M were spotted on media with the indicated compounds and monitored for growth.
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