(
A) Using the assay system described in
Figure 4E, wild-type,
hta1Δ,
hta2Δ, and
hta1-T126A strains were subjected to DSB induction at the
leu2 cut site (leu2::cs) by growth in galactose for the indicated times (hours in 2% galactose). Genomic DNA was purified, restriction digested, separated by gel electrophoresis and Southern blotted as previously described (
Vaze et al., 2002). The timing of HO cutting (HO cut band) and repair product formation via SSA (SSA product) is not different between the indicated genotypes. (
B) Using the SSA assay described in
Figure 4E, Rad53 phosphorylation profiles were determined by Western blot in the WT and
hta1Δ strains. The
hta1Δ strain shows no significant difference from WT in the kinetics of Rad53 phosphorylation and dephosphorylation after HO cutting and repair.