Figure 1.

Recruitment of rad1^nuc- to 3′ non-homologous tail carrying recombination intermediates. (A) Domain structure of Rad1 and the location of rad1 mutations that disrupt nuclease activity. (B) Schematic illustration of HO break and the flanking repeats in tNS1379/EAY1141 or YMV80 strain used for ChIP assay. Location of primers and the size of the repeats are shown. Paired arrows indicate primers used for ChIP assay. (C, D) The requirements of Rad10, Rad52, Msh2, Saw1, and Slx4 for the recruitment of rad1^nuc- to the proximal (pJC1 and pJC2) (C) or distal (pJC3 and pJC4) (D) side of the 3′-flap site in tNS1379 strain. Association of rad1^nuc- to the 3′-flap 6 h post HO induction in the msh2Δ derivative of YMV80 strains is also shown (D). Fold enrichment represents the ratio of the rad1 IP PCR signal before and after HO induction, normalized by the PCR signal of the MAT control. Data represent the mean±s.d. of three or more independent experiments.