TABLE 3.
Fidelity of WT and mutant pol η on TT dimer by steady-state kinetics
| Enzyme | Template:dNTPa | Km (μM) | kcat (min−1) | kcat/Km (μM−1 min−1) | fincb |
|---|---|---|---|---|---|
| S. cerevisiae pol η WT | 3′-T:dATP | 2.1 ± 1.3 | 7.5 ± 0.42 | 4.4 ± 1.9 | 1 |
| 3′-T:dGTP | 130 ± 14 | 2.3 ± 0.16 | 0.018 ± 0.0029 | 4.0 × 10−3 | |
| 3′-T:dCTP | 330 ± 160 | 0.37 ± 0.13 | (1.2 ± 0.15) × 10−3 | 2.6 × 10−4 | |
| 3′-T:dTTP | 210 ± 83 | 1.8 ± 0.16 | (9.8 ± 5.5) × 10−3 | 2.2 × 10−3 | |
| 3′-CPD:dATP | 6.0 ± 2.3 | 5.7 ± 0.76 | 1.1 ± 0.46 | 0.24 | |
| S. cerevisiae pol η F34L | 3′-T:dATP | 460 ± 43 | 2.6 ± 0.30 | (5.7 ± 0.14) × 10−3 | 1 |
| 3′-T:dGTP | 1,100 ± 150 | (7.5 ± 0.80) × 10−3 | (7.1 ± 0.53) × 10−6 | 1.3 × 10−3 | |
| 3′-T:dCTP | NDc | ND | ND | ND | |
| 3′-T:dTTP | ND | ND | ND | ND | |
| 3′-CPD:dATP | 600 ± 140 | 0.16 ± 0.023 | (2.8 ± 0.33) × 10−4 | 0.048 |
a
Template and dNTP represent template base and incoming dNTP, respectively. The name of each template site is schematically illustrated in Fig. 1.
b
finc, kcat/Km value relative to that for T:dATP (which was set at 1.0).
c
ND, incorporation was below the detection limit and thus was not detected.