Figure 3
From: Hypersusceptibility mechanism of Tenofovir-resistant HIV to EFdA
ATP- and PPi-dependent rescue of EFdA-MP terminated primers by WT and K65R RTs. (A) ATP-dependent rescue of Td31/Pd18-P0-EFdA-MP. Purified Td31/Pd18-P0-EFdA-MP was incubated with WT or K65R RT in the presence of 10 mM MgCl2, 3.5 mM ATP, 100 μM dATP, 0.5 μM dTTP, and 10 μM ddGTP at 37°C. Aliquots of the reaction were stopped at the indicated time points (0–90 min). The results of at four independent experiments were plotted using one site hyperbola in Graphpad Prism 4. (B) PPi-dependent rescue of Td31/Pd18-P0-EFdA-MP. Purified Td31/Pd18-P0-EFdA-MP was incubated with WT or K65R RT in the presence of 6 mM MgCl2, 150 μM PPi, 100 μM dATP, 0.5 μM dTTP, and 10 μM ddGTP at 37°C. Aliquots of the reaction were stopped at the indicated time points (0–40 min). The results of two independent experiments were plotted using one site hyperbola in Graphpad Prism 4.