Fig. 5: Ubiquitination requirement for RNF168 selectivity on H2A variants.

a Superimposition of H2A-containing, H2AZ-containing, and macroH2A-containing nucleosome illustrates the detailed residues required for RNF168-mediated site-specific ubiquitination. Zoom on the acidic patch (red), alpha1-extension helix region (blue) and RNF168-targeted lysine residues (green). b Proximity restriction of RNF168-mediated site-specific ubiquitination on H2A. Sequence alignment of human H2AX and yeast HTA (yHTA) at the N-terminal tail. The N-terminal sequence of the yHTA K13m mutant (ΔASQ) used as in c. The alpha1-extension helix residues are highlighted in blue. The proximal lysine residues are highlighted in green. c SFB-yHTA and mutant were co-transfected with Myc-RNF168 in HEK293T cells for 24 h and analyzed by western blot. Repeated three times independently with similar results. d Electrostatic potential (red-negative, blue-positive) analyses of H2A variant-containing nucleosomes. Zoomed illustration of the N-terminus and C-terminus of H2A variants. Histone tail lysines were labeled. H2AZ C-terminal lysines were absent in the illustration and G119 residue was marked. Source data are provided as Source Data file.