Table 3.

Steady-state kinetic parameters for nucleotide incorporation opposite the 5′ T of the (6-4) TT photoproduct by yPolζ

TemplatePrimer:template base pairIncoming residueVmax (nM/min)Km (μM)Vmax/Km Relative efficiencya
NondamagedG·TG0.11 ± 0.01170 ± 330.00066.0 × 10−4
A0.70 ± 0.050.67 ± 0.061.01
T0.38 ± 0.04130 ± 80.00292.9 × 10−3
C0.12 ± 0.07250 ± 560.00055.0 × 10−4
A·TG0.13 ± 0.01140 ± 110.00091.0 × 10−4
A1.06 ± 0.070.12 ± 0.078.81
T0.42 ± 0.0294 ± 140.00455.1 × 10−4
C0.34 ± 0.05140 ± 220.00242.7 × 10−4
(6-4) TTG·TG0.13 ± 0.01130 ± 170.0012.6 × 10−4
A1.2 ± 0.030.32 ± 0.053.81
T0.34 ± 0.08180 ± 320.00195.0 × 10−4
C0.27 ± 0.03160 ± 240.00174.5 × 10−4
A·TG0.24 ± 0.0257 ± 100.00423.0 × 10−3
A0.89 ± 0.040.63 ± 0.111.41
T0.43 ± 0.02130 ± 100.00332.4 × 10−3
C0.28 ± 0.03200 ± 340.00141.0 × 10−3
  • a For each DNA substrate, the relative efficiency of nucleotide incorporation was determined by dividing the efficiency (V max/Km ) for incorporating each nucleotide by the efficiency (V max/Km ) for incorporating the correct nucleotide, A.