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. 2014 Oct 28;42(20):12722–12734. doi: 10.1093/nar/gku1027

Figure 5.

Figure 5.

Infidelity of manganese-dependent 8-oxo-dGMP and 8-oxo-GMP incorporation. (A) Polymerase reaction mixtures containing 10 mM Tris-HCl, pH 7.5, 50 nM 5′ 32P-labeled 13-mer/18-mer primer-template as specified, 1 μM DinB2, 1 mM MnCl2, and either 100 μM oxo-dGTP or oxo-rGTP were incubated at 37°C for 10 min. The reaction products were analyzed by urea-PAGE and visualized by autoradiography. (B) Kinetics. Primer extension reactions were constituted as in (A). Aliquots were withdrawn at the times specified and quenched with EDTA/formamide. The % primer extension with oxo-dGTP (left panel) or oxo-rGTP (right panel) is plotted as a function of reaction time for each oligo-dX5 template. Each datum is the average of three separate experiments ±SEM. The rate constants (kobs ± SE) for the first step of oxo-dGMP addition to the primer-template were obtained by non-linear regression curve fitting of the data to a one-phase association function in Prism.