(A) Southern blots of pulsed-field gels, probed with a left end–adjacent probe (nt 15,838–16,857) to detect DSBs on chromosome III in a normal chromosome or one with all three active replications origins on chr III-L deleted (arsΔ). DNA was prepared from sae2Δ (MJL2306), sae2Δ arsΔ (MJL2529), dmc1Δ (MJL2560), and dmc1Δ arsΔ (MJL2683) 6 h after initiation of meiosis. Quantification traces for each lane, normalized to total lane intensity, are shown (normal chr III-L, black; arsΔ, gray). DSB frequencies in the left arm (I + II), central “cold” region (III), and right arm (IV + V) domains [32] and frequencies of uncut chromosomes are expressed as fraction of total lane signal. Values are corrected to account for double cutting events (see Materials and Methods). The uncorrected values are as follows: for interval I + II: sae2Δ, 0.27; sae2Δ arsΔ, 0.07; dmc1Δ, 0.42; dmc1Δ arsΔ, 0.33. For interval III: sae2Δ, 0.02; sae2Δ arsΔ, 0.02; dmc1Δ, 0.13; dmc1Δ arsΔ, 0.11. For interval IV + V: sae2Δ, 0.3; sae2Δ arsΔ, 0.4; dmc1Δ, 0.24; dmc1Δ arsΔ, 0.3.
(B) Frequencies of crossing-over in the indicated intervals (centiMorgans ± standard error), in wild type (MJL3237) or strains with an arsΔ chrIII-L (MJL3236). Black circle indicates centromere; white squares indicate replication origins (ARS305, ARS306, ARS307) that are deleted in replication-delayed chromosome arm.