Figure 3 : Gcn5 promotes HR and suppresses NHEJ.

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

Figure 3

(a,b) Methylation and acetylation of H3K36 are mutually inhibitory. (a) Western blot analysis of H3K36me3 in wild type (Wt), set2Δ and gcn5Δ nuclear extract. (b) Western blot analysis of H3K36ac in wild type (Wt), set2Δ and gcn5Δ nuclear extract. (c) Fivefold serial dilutions of wild-type (Wt), gcn5Δ and rad51Δ strains on YE6S and YE6S+5 μg ml−1 bleomycin. Plates were incubated at 32 °C for 3 days. At least two biological replicates were performed. (d) Percentage DSB-induced marker loss in wild type (Wt) and gcn5Δ containing Ch16-RMYAH. The levels of NHEJ/SCC, GC, Ch16 loss and LOH are shown. Means±s.e. of three experiments are shown. * represents significant difference compared with wild type (P<0.05, t-test). See also Table 1. (e) Quantification of Rad51-CFP foci in wild-type and gcn5Δ strains following exposure to 50 Gy IR. (f) Quantification of Rpa1(Rad11)-GFP foci in wild-type and gcn5Δ strains strains following exposure to 50 Gy IR treatment. Data are the mean of three experiments and error bars (±s.e.) are shown. (g) Gcn5 suppresses NHEJ plasmid rejoining. The percentage of leu+ colonies obtained following transformation and rejoining of a LEU2 plasmid (pAL19) linearized with EcoRI, PstI into wild-type, lig4Δ and gcn5Δ strains compared with uncut plasmid control is shown. Means±s.e. of three experiments are shown. (h) Gcn5 functions redundantly with other HAT complex subunits in the DNA damage response. Fivefold serial dilutions of wild-type (Wt), gcn5Δ, hat1Δ, hat1Δ gcn5Δ, nto1Δ, nto1Δ gcn5Δ, eaf6Δ, eaf6Δ gcn5Δ, eaf7Δ, and eaf7Δ gcn5Δ strains on YE6S and YE6S+5 μg ml−1 bleocin. At least two biological replicates were performed.