FIG. 5.

Formation of meiotic DSBs in the tdh1⁺ locus depends on Rec12 and the Atf1-Pcr1 complex. (A) Haploid pat1-114 rad50s strains PKH138 (wild type [WT]), PKH118 (rec12Δ), PKH163 (atf1Δ), and PKH160 (pcr1Δ) were cultured, and DNA was prepared as described in Materials and Methods. tdh1⁺ ORF and CRE sequences are indicated as in Fig. 2. An arrowhead indicates the DSB site. The genome-wide DSBs in the same samples were analyzed by pulsed-field gel electrophoresis. (B) DNA samples from the MNase-digested chromatins and synchronous meioses in Fig. 1 were digested with ApaLI and AflII and analyzed in the same gel. DSBs are indicated by a dotted line; tdh1⁺ ORF and CRE sequences are indicated as in Fig. 2.