Saccharomyces cerevisiae DNA Ligase IV Supports Imprecise End Joining Independently of Its Catalytic Activity
Figure 4
Catalytically inactive Dnl4 protein does not impede DSB resection.
DSBs were induced in yeast strains bearing the ILV1-cs allele similarly to Figure 3 and monitored by Southern blotting. (A) Blot showing that DSB resection is not observed without the addition of glucose at 60 min. (B) Example blot showing formation of the HO-cut band and its subsequent disappearance by a combination of NHEJ and DSB resection. (C) and (E) The ratio of the HO-uncut band to the APN1 control was normalized to the ratio at time 0 to allow monitoring of DSB formation and repair by NHEJ. (D) and (F) The ratio of the HO-cut band to the APN1 control was normalized to the ratio at 60 min when DSB formation was maximal. Disappearance of the HO-cut band at subsequent times results from NHEJ (wild-type strain only) and/or DSB resection. (C) and (D) show results from asynchronous cells while (E) and (F) show results from cells arrested in G1 with α-factor. Results are the mean ± standard deviation of five independent experiments.