Fig. 5: Additional SLX4 segments resolved in the structure of the XPF-ERCC1-SLX4IP-SLX4330-555-DNA complex.
From: Molecular basis of XPF-ERCC1 targeting to SLX4-dependent DNA repair pathways
a Left: Cryo-EM map of the XPF-ERCC1-SLX4IP-SLX4330-555-DNA complex. Three SLX4 segments (orange) are visualised and labelled. Right: Cryo-EM map of the DNA-free complex from the same data collection shown in the same orientation. Only SLX4 residues 526-552 are visualised, in line with the higher-resolution reconstructions shown in Fig. 1. Maps are shown without any b-factor sharpening applied to facilitate visualisation of weaker densities. b Schematic depiction of the sequence elements and domains contained in the SLX4 constructs used for in vitro endonuclease assays of XPF-ERCC1-SLX4IP-SLX4 complex variants. c In-vitro endonuclease assay comparing the activity of XPF-ERCC1 to the activities of different variants of the XPF-ERCC1-SLX4IP-SLX4330-555 complex. Conversion of the input substrate (see Fig. 3a) into 23-nt product was monitored by detection of Cy3 fluorescence. To facilitate visualisation, 10 min time points are marked with a red dot. The removal of the UBZ-2 domain and the addition of the BTB domain do not have an appreciable effect on endonuclease activity. Removal of the sequence elements that are solely visualised in the DNA-bound XPF-ERCC1-SLX4IP-SLX4330-555 complex strongly reduces cleavage activity. Two additional experimental repeats and all protein sample processing controls are provided in Supplementary Fig. 7a–e. Source Data are provided as a Source data file.
