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. 2014 Mar 20;42(9):5830–5845. doi: 10.1093/nar/gku214

Figure 8.

Figure 8.

Processivity of the polymerase activity of PrimPol. PrimPol was pre-incubated for 30 min at 37°C with an undamaged DNA primer-template substrate to allow PrimPol to bind to the DNA. The reaction was initiated by the addition of dNTPs and an excess of sonicated herring sperm DNA (trap) and time points taken at 15, 30, 60, 120 and 360 s. (A) After 360 s, wild-type PrimPol incorporated up to 4 nucleotides opposite the template but a significant fraction of enzyme incorporated only 1, 2 or 3 nucleotides (left panel). To confirm that the trap prevents polymerase extending from a second template, the trap was also added into the pre-incubation mix with PrimPol and the DNA substrate. This reaction was supplemented with dNTPs and there is no extension (right panel), thus successfully exhibiting the effectiveness of the trap. (B) PrimPol1–354 also predominantly incorporates up to 4 nucleotides but there were fewer polymerases incorporating only 1, 2 or 3 nucleotides (left panel). The effectiveness of the trap was also successfully confirmed (right panel). (C) Percentage of PrimPol molecules incorporating at least n dNTPs for either full-length PrimPol or PrimPol1–354, calculated using Equation (1).