Figure 4. Role of Top3 and RPA in resection.
a, Top3-Rmi1 and top3-Y356F-Rmi1 were tested in the resection system with 3′-labeled substrate. Symbol: Bl, no protein added.
b, Analysis of 5′ resection in top3Δ cells supplemented with the low copy pRS315 vector carrying TOP3 or top3-Y356F gene. Analysis was done as in Figure 2d.
c, Crossover (CO’s) and noncrossover (NCO’s) recombinants are distinguished based on their size (i). The frequencies of CO’s 8 hr after break induction were determined7 and the mean values ±SD from three independent experiments were plotted (ii).
d, SSB or hRPA supports DNA unwinding but not end resection. Symbol: HD, heat denaturation of DNA.
e, Yeast RPA stimulates 5′ end digestion (i) but attenuates 3′ end digestion (iii) by Dna2. The mean values ±SD from three independent experiments were plotted in (ii) and (iv).