Figure 5.

XL-MS Experiments to Probe the Conformational Changes in UHRF1 upon Chromatin and E2 Binding

(A–E) XL-MS diagrams of UHRF1 in isolation (A), with mono-nucleosomes (B), 12 × 187 bp chromatin arrays (C), UbcH5a/UBE2D1 (D), and in the context of 12 × 187 bp arrays and UbcH5a (E). Intra-protein crosslinks are in red and inter-protein crosslinks in blue (histones) or green (UbcH5a). XL-MS experiments with UHRF1^WT are on the left and UHRF1^ΔMW on the right. Schematic representations of the proposed UHRF1 conformation in each experiment are shown with arrows indicating the predominant crosslinks from UHRF1 to histones and UbcH5a. UHRF1 domains are color coded as in Figure 1A, with histones in yellow and UbcH5a in green. Table S1 has the list of crosslinks identified in each experiment, and Data S1 has an example spectrum as output from StavRox.