Figure 4.

Effect of Rev1 protein on Polζ-catalyzed nucleotide insertion opposite the AAF-dG adduct. A 17mer primer was labeled with ³²P at its 5′ end and annealed to the damaged template with the primer 3′ end terminating right before 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). 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.