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. 2013 Dec 5;9(12):e1003814. doi: 10.1371/journal.ppat.1003814

Figure 3. 5-OH-dCTP is an in vivo substrate of mycobacterial MazG proved by chemical genetic analysis.

Figure 3

The Msm competent cells were prepared as described in Materials and Methods , the nucleotides were incorporated by transformation. (A) mazG-null Msm (ΔmazG) exhibited a dose-dependent 5-OH-dCTP induced mutagenesis. wt, wild-type Msm. The data shown are mean ± S.E. of four repeats. (B) The rifampicin-resistance mutation frequencies of wild-type Msm, the mazG-null mutant and the complemented mutant treated with 5-OH-dCTP and dCTP. Nucleotides were added at 100 µM final concentration. Mean ± S.E of 12 independent transformations. **p<0.01 vs wt, * p<0.05 vs wt. (C) The rifampicin-resistance mutation frequencies of wild-type and mazG-null Msm treated with 100 µM 2-OH-dATP, 5-CHO-dUTP and normal dNTPs. Mean ± S.E of 8 independent transformations. (D) mazG-null Msm is susceptible to killing by 5-OH-dCTP. Strains were treated with 100 µM 5-OH-dCTP for 5 hours at 37°C. Shown is percent survival compared to an untreated control (100%). The data shown are mean ± S.E. of four repeats.