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. 2015 Sep 30;35(21):3714–3725. doi: 10.1128/MCB.00298-15

FIG 6.

FIG 6

Loss of the methyl-binding domain of Set3 suppressed esa1 DNA damage sensitivity. (A) The esa1 set1Δ strain was synthetically sick. The esa1 set1Δ strain grew slowly at 30°C on rich medium and was inviable when grown on 0.2% DMSO or 10 μg/ml of CPT. (B) Mutation of H3K4 to alanine disrupted suppression of esa1 DNA damage sensitivity by hos2Δ, which was also sensitive to H3K4A. Shown are serial dilutions of wild-type (LPY14161), esa1-414 (LPY14163), esa1-414 hos2Δ (LPY15906), and hos2Δ strains expressing wild-type histones from a plasmid; wild type (LPY21480), esa1-414 (LPY21481), esa1-414 hos2Δ (LPY21482), and hos2Δ (LPY21483) strains expressing H3K4A from a plasmid. (C) esa1 strains are synthetically sick when combined with COMPASS complex deletions promoting simultaneous loss of H3K4 di- and trimethylation. The following strains were assayed: wild-type (LPY6497), esa1-531 (LPY14757), esa1-531 cps25Δ (LPY21498), esa1-531 cps40Δ (LPY21495), esa1-531 cps60Δ (LPY21503), esa1-531 hos2Δ (LPY14761), cps25Δ (LPY21499), cps40Δ (LPY21494), and cps60Δ (LPY21520) strains. (D) hos2Δ could not suppress esa1 when CPS25 or CPS40 were also deleted. Strains tested included wild-type (LPY6497), esa1-531 (LPY14757), esa1-531 hos2Δ cps25Δ (LPY21656), esa1-531 hos2Δ cps40Δ (LPY21661), and esa1-531 hos2Δ (LPY14761) strains. (E) Vector-transformed (pLP1358) wild-type, esa1-531, esa1-531 hos2Δ, esa1-531 set3Δ, set3Δ, and hos2Δ strains shown in Fig. 5B and Fig. S3 in the supplemental material were compared to set3-W140 (PHD domain mutant, pLP3020)-transformed esa1-531 set3Δ and set3Δ strains.