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Fig. 2 | Epigenetics & Chromatin

Fig. 2

From: The DNA repair protein SHPRH is a nucleosome-stimulated ATPase and a nucleosome-E3 ubiquitin ligase

Fig. 2

SHPRH is a nucleosome-stimulated but not a dsDNA-stimulated ATPase. ATPase assay. Increasing concentrations of enzymes (hSWI/SNF: 465 ng and 930 ng in a final volume of 15 μl or SHPRH: 57 nM and 170 nM) were incubated in the presence of buffer only (white bars), 5 kb plasmid DNA (200 ng; gray bars) or the same plasmid assembled into nucleosome arrays (200 ng; black bars) and 20 μM ATP/60 μM Mg2+ supplemented with traces of γ-[32P]ATP. Hydrolyzed phosphates and non-hydrolyzed ATP were separated using thin layer chromatography after incubating the reactions for 40 min at 37 °C. The percentage of ATP hydrolysis was quantified using a phosphorimager. Error bars reflect standard deviation derived from two independent experiments

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