Figure 2.

Deletion of SET1 Is Lethal in glc7-127 Cells

1. Schematic representation of the plasmid shuffle approach used to combine the glc7-127 mutation or the sds22-6 mutation with SET1 deletion. A URA3-marked plasmid bearing the wild-type GLC7 gene was introduced into either glc7-127 or sds22-6 cells. The SET1 gene was then disrupted. Growth of the resulting cells was compared on rich media (YPD) or on 5-FOA-containing media, which selects for cells that have lost the URA3-marked GLC7 plasmid.
2. Serial dilutions (10-fold) of the indicated strains reveals that deletion of SET1 is synthetic lethal with glc7-127 in the absence of the exogenous wild-type GLC7 allele on 5-FOA-containing media (at 25°C).
3. Yeast strains with the indicated genotypes were serially diluted 10-fold, spotted onto YPD medium or 5-FOA medium and then grown at indicated temperatures for 3 days. Note the enhanced temperature sensitivity of the sds22-6 set1 Δ cells on 5-FOA-containing media at 30°C and on rich media at 33°C relative to either sds22-6 or set1 Δ single mutants.
4. Yeast strains with the indicated genotype were again serially diluted 10-fold, spotted onto YPD medium, and then grown at indicated temperatures for 2 days. As expected, the glc7-127 mutation suppresses the ipl1-2 temperature-sensitive phenotype at 32.5°C. Importantly, ipl1-2 glc7-127 set1 Δ triple-mutant cells are viable, and the temperature-sensitive phenotype of ipl1-2 is also suppressed in these cells.