Figure 2 : Set2 is required for C-NHEJ.

From: A histone H3K36 chromatin switch coordinates DNA double-strand break repair pathway choice

Figure 2

(a) Serial dilutions of the indicated strains were spotted onto YE6S containing 5 μg ml−1 bleomycin or no drug. Plates were incubated at 32 °C for 3 days. (b) Percentage survival of strains indicated compared with wild-type following plating on YE6S +/− the indicated bleocin concentration. Plates were incubated for 3 days at 32 °C and then scored. Means±s.e. of four independent experiments are shown. (c) Set2 is required for NHEJ plasmid rejoining. The percentage of leu+ colonies obtained following transformation rejoining of a LEU2 plasmid (pAL19) linearized with EcoRI, PstI or PvuII following transformation into wild-type (Wt), lig4Δ and set2Δ strains compared with uncut plasmid control is shown. Means±s.e. of three experiments are shown. See also Supplementary Fig. 2. (d) Quantification of Rpa1(Rad11)-GFP foci in wild-type (Wt) and set2-R255G strains following exposure to 50 Gy IR treatment. Mean±s.e. of three experiments are shown. See also Supplementary Fig. 5. (e) DSB induction results in a Set2-dependent increase in histone H3K36me3 levels proximal to the HO break. qChIP enrichments of H3K36me3 in wild-type (Wt) or set2Δ cells at various times (60, 90 or 120 min) following urg-HO induced DSB induction at the MATa site between SPAC3H1.10 and hsr1 on Chromosome I, as previously described59. Enrichment of immunoprecipitated DNA at 50 bp from the break relative to that at act1 is presented, as a ratio of that observed in a strain expressing no HO endonuclease. Error bars represent s.d. from at least two biological replicates. (f) DSB induction results in a Set2-dependent increase in Ku80 levels proximal to the HO break. qChIP enrichments of Ku80-myc in wild-type (Wt) or set2Δ cells at various times (60, 90 or 120 min) following urg-HO induction as described above. Enrichment of immunoprecipitated DNA at 50 bp from the break, relative to that at the fbp1 locus is presented. Error bars represent s.d. from at least two biological replicates.