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Figure 2

From: Arabidopsis thaliana PrimPol is a primase and lesion bypass DNA polymerase with the biochemical characteristics to cope with DNA damage in the nucleus, mitochondria, and chloroplast

Figure 2

AtPrimPol recognizes a single-stranded DNA sequence and preferentially incorporates dNTPs over rNTPS. (A) AtPrimPol recognizes the cryptic G. (A) 24% denaturing gel electrophoresis showing AtPrimPol primer synthesis assayed on four different templates where the base corresponding to the cryptic nucleotide was systematically changed (lanes 4 to 11). Reactions were carried out in the presence of ATP and GTP or dGTP. Primer synthesis was mainly observed in reactions contained dGTP and longer products were only synthesized on a template containing the 3′-GTC-5′ motif. A control reaction showing primer synthesis (pAC and pACA products) by T7 Primase is used as molecular weight marker (lane 3). (B) AtPrimPol prefers rATP in its 5′ position. Primer formation was evaluated on four templates containing the 3′-GAC-5′, 3′-GCC-5′, 3′-GGC-5′, and 3′-GTC-5′ sequences. Each of these templates were assayed with U/T, G/dG, C/dC and A/dA respectively at 100 μM, and 100:1 μM. Primer synthesis on templates containing the CAG and CCG sequences was inefficient, whereas templates containing the 3′-GGC-5′ and 3′-GTC-5′ sequences readily primed and extended beyond the length of the template. Original data used to compose this figure is present in Fig. S9.

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