Fig. 4.

RFC and Elg1 stimulate the Mus81 complex and Rad27 nuclease activities. a Reaction mixtures containing DNA substrate (5 nM) and the Mus81 complex (M/M) (0.25 nM, lanes 1–5) or Rad27 (0.25 nM, lanes 6–10) were incubated with increasing amounts of the RFC complex (0.25, 0.5, 1.25, and 2.5 nM) at 37 °C for 20 min and then analyzed. b Quantification of data from three independent experiments. c RFC alone does not cleave the DNA substrate. Time-course enhancement of the Mus81 complex activity by RFC. The Mus81 complex (0.25 nM) was incubated with the 3′ flap DNA substrate (5 nM) at 37 °C for 60 min in the presence of RFC (1.25 nM). Aliquots of the reactions were taken at the indicated times and analyzed. Last lane indicates the effect of RFC complex on the DNA substrate in the absence of the Mus81 complex. d, e Time course enhancement of the Mus81 complex and Rad27 cleavage by RFC. The Mus81 complex (M/M, 0.25 nM) and Rad27 (0.25 nM) were incubated with the 3′ flap or 5′ flap DNA substrates (5 nM) at 37 °C for 30 min (d) in the absence or (e) in the presence of RFC (1.25 nM). Aliquots of the reactions were taken at the indicated times and analyzed. f Quantification of data in (d) and (e) from three independent experiments. g Reaction mixtures containing DNA substrate (5 nM), Mus81-Mms4 (0.25 nM, lanes 1–5) or Rad27 (0.25 nM, lanes 6–10) and increasing amounts of Elg1-RFC complex (0.25, 1.25, 2.5, and 10 nM) were incubated at 37 °C for 20 min and then analyzed. h Quantification of data in (g) from three independent experiments. *Raw data provided in Additional file 2