Figure 7.

Specificity of nucleotide incorporation during extension synthesis from opposite the AAF-dG adduct by yeast Polζ with or without Rev1. Four 18mer primers that differed by 1 nt at the 3′ end were labeled with ³²P at their 5′ ends and separately annealed to the damaged template with the primer 3′ end opposite the AAF-dG adduct as shown on the top. DNA polymerase assays were then performed with 10 ng (50 fmol) of yeast Polζ in the presence of all four dNTPs (N₄), or a single deoxyribonucleotide triphosphate dATP (A), dCTP (C), dGTP (G) or dTTP (T) as indicated. Lanes 1–5, reactions with Polζ alone; lanes 6–10, reactions with Polζ plus purified yeast Rev1 protein (5 ng, 45 fmol). DNA size markers in nucleotides are indicated on the left. (A) C opposite the lesion; (B) A opposite the lesion; (C) T opposite the lesion and (D) G opposite the lesion.