Schematic of the H‐0 BIR assay. DSB is induced at a modified MATa locus (Chr. III). Strand invasion occurs within the “Z” sequence (blue box) and DNA synthesis continues to the end of the chromosome.
Representative Southern blots showing DSB repair products in WT, rad59Δ, rad52‐R70A, and rad59Δ rad52‐R70A cells. DNA was digested with Bsp1286I and probed with a MAT‐distal sequence (yellow box in panel A).
Viability of indicated strains is shown (mean ± SD; n = 3).
Repair efficiency compared to parental MATa at time 0 h (left) and repair efficiency of indicated mutants compared to WT by 6 h (right) (mean ± SD; n ≥ 3).
Representative Southern blots showing BIR repair product in H‐0 system in WT and indicated mutant cells.
Repair efficiency compared to parental MATa at time 0 h (left) and repair efficiency of indicated mutants compared to WT by 6 h (right) (mean ± SD; n ≥ 3).
Model presenting the possible function of DNA binding/annealing domain of Rad52 in stabilizing the D‐loop. Rad52 stabilizes/extends D‐loop by three‐strand exchange (left) or Rad52 anneals a 3′ invading strand unwound from its template back to disrupted, but still RPA‐bound, D‐loop (right).