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. 2019 Feb 4;15(2):e1007956. doi: 10.1371/journal.pgen.1007956

Fig 1. Temperature-dependent dynamic deactivation of pol V Mut E38K/ΔC17.

Fig 1

(A) Sketch showing dynamic deactivation of pol V Mut E38K/ΔC17 (200 nM) with ATPγS /ATP (500 μM) at 37°C and 30°C in the presence of saturating concentration of dNTP’s (mix of dTTP, dCTP, dGTP 500 μM each). Representative DNA synthesis gels for pol V Mut E38K/ΔC17 with ATP/ATPγS at 37°C and 30°C are presented in (B-E). Each experiment was repeated 3 times, and the average % PE (percent p/t DNA extended) with the SD (standard deviation) for each reaction time point is graphed in panels (F-G). Pol V Mut E38K/ΔC17 deactivates in about 1.5 h at 37°C and completes only one round of DNA synthesis with ATP/ATPγS (B-C and F-G black circles in the graphs). In contrast, at 30°C, pol V Mut E38K/ΔC17 deactivates in about 3h, which allows the enzyme to complete 4 rounds of DNA synthesis with ATPγS and 3 rounds with ATP (D-E and F-G white triangles in the graphs). Deactivated pol V Mut E38K/ΔC17 is not “dead” and is reactivated by adding RecA* (200 nM; RecA* in B-G). % PE refers to percent p/t DNA extended and was calculated as the integrated gel band intensities of extended hairpin DNA over total DNA intensity.