Figure 1: The H2A/H2B dimer is specifically modified at K13–15 by the RNF168 RING domain.
From: The nucleosome acidic patch plays a critical role in RNF168-dependent ubiquitination of histone H2A
(a) RNF168 targets H2A in isolation in an inefficient manner and its activity is not specific for K13–15. RING1B/BMI1 are inactive towards the histone protein alone. Ubiquitination assay performed in presence of H2A WT or K13–15R in isolation. (b) RNF168 targets H2A specifically on K13–15 within the H2A/H2B dimer. Ubiquitination assays performed in presence of WT or K13–15R H2A in H2A/H2B dimer. Single time-point assay, where RING1B/BMI1 were used as negative control. Time course assay (10–30–60–90 min) in the presence of WT or K13–15 R dimers (assays performed at 200 mM NaCl). (c) RNF168 has comparable activity for H2A in dimers and in nucleosomes, while isolated H2A is targeted with low efficiency. Time course assay (10–30–60–90 min) to compare activity of RNF168 RING domain towards H2A alone, H2A/H2B dimers and NCPs.
