Extended Data Fig. 6: Monoubiquitination assays in the absence of DNA suggest that the three phosphomimetic mutations have an additive effect on D2-I closure.
From: The DNA-damage kinase ATR activates the FANCD2-FANCI clamp by priming it for ubiquitination
(a) Time-course monoubiquitination assays in the absence of DNA were performed with FANCD2 incubated with different FANCI constructs and analyzed using SDS-PAGE. Eight different FANCI constructs were used: wild-type FANCI, FANCI-Dxx (S558D), FANCI-xDx (S561D), FANCI-xxD (T567D), FANCI-DDx (S558D and S561D), FANCI-DxD (S558D and T567D), FANCI-xDD (S561D and T567D), FANCI-3D (S558D, S561D and T567D), and FANCI-delta loop (deletion of residues 555–582). (b) Quantification of monoubiquitination assays performed in panel a. Data obtained from four independent assays were quantified as percentage of ubiquitinated FANCD2. Replicate data were fitted using a global non-linear regression function and plotted. Data points of each replicate (symbols), the corresponding means (solid lines) and the 95% confidence intervals (shaded area between dashed lines) are shown.
