DNA polymerases eta and kappa and benzo[a]pyrene diol epoxides-2 mutagenesis

Polymerase fidelity assays were used to assess human DNA polymerases eta and kappa ability to incorporate nucleotides, correct or incorrect, on non-adducted 16-mer templates and on the same templates containing (+) and (-) anti-benzo[a]pyrene diol epoxide adducts. In assays with 1mM dNTPs, polymerase eta was found to incorporate A and G far more than T or the correct C. Vmax/Km for DNA polymerase eta incorporation of different nucleotides on a non-adducted dA template ranged between 10-8 to 10-6 nM primer/muM dNTP/min and on a non-adducted dG template ranged between 10-6 to 10-5 nM primer/muM dNTP/min. DNA polymerases kappa Vmax/Km values for misincorporation on a non-adducted dA template ranged from 10-6 to 10-5 nM primer/muM dNTP/min and on a template containing a trans R BaP DE-2 incorporation of G was found to be 1.65 x 10-6 nM primer/mM dNTP/min and for A 1.33 x 10-6 nM primer/mM dNTP/min.