Figure 2.

DNA interaction and activation of ARTD2_FL and the different point mutants. (A) The specific DNA dependent activity of full length ARTD2_FL and point mutants were measured using fluorescence activity assay. Protein (50 nM) was incubated with 5 μM NAD⁺ for 1 hour and leftover NAD⁺ was quantified. DNA was used in 2:1 protein:DNA ratio. Data are presented as means and standard deviation of the quadruplicates. Each DNA was assayed with all the mutants and data is shown for the ones having any NAD⁺ consumption. (B) Interaction of ARTD2_FL and the different point mutants were studied using EMSA. Residues on the 3′ side of the break are labeled with 3′, residues on the 5′ side of the break are labeled with 5′ and residue interacting with a nucleobase is labelled with **.