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. 2015 Jan 27;43(3):1684–1699. doi: 10.1093/nar/gkv025

Figure 5.

Figure 5.

The importance of the N21–26 motif for the interaction of Mus81-NF120 with Rad27. (A) GST pull-down assays with GST-Rad27 and derivatives of Mus81-NF120 (WT) that include Mus81-NF120Δ21–22 (Δ21–22), Mus81-NF120Δ21–24 (Δ21–24) and Mus81-NF120Δ21–26 (Δ21–26) (see text for details). (B) SDS-PAGE (8%) analysis of purified recombinant Mus81Δ21–22–Mms4Δ40N, Mus81Δ21–24–Mms4Δ40N and Mus81Δ21–26–Mms4Δ40N complexes. The gel was Coomassie-blue stained. The sizes of molecular mass marker are indicated in kDa. (C) Comparison of endonuclease activities of three recombinant Mus81Δ21–22–Mms4Δ40N, Mus81Δ21–24–Mms4Δ40N and Mus81Δ21–26–Mms4Δ40N complexes. Reactions were carried out in standard reaction mixtures containing 15 fmol of 3′F substrate and increasing amounts (5, 10, 20 and 40 fmol) of Mus81 complexes. The amount (fmol) of cleavage products formed by the endonuclease activity of Mus81 complexes on 3′F substrate was plotted against the amount (fmol) of Mus81 complexes added. The graph with error bars indicated was obtained from four independent experiments. The error bars represent the standard deviation from the mean of four independent experiments. (D) The influence of Rad27 on the endonuclease activities of Mus81Δ21–22–Mms4Δ40N, Mus81Δ21–24–Mms4Δ40N and Mus81Δ21–26–Mms4Δ40N complexes. Reactions were carried out and the graph obtained was presented as described in Figure 3C.