| Â | cGTP incorporation | ncGTP misincorporation |
|---|
RNAP | kpol (s-1) | Kd (μM) | kpol (s-1) | Kd (μM) |
|---|
WT/Stl | 3.6 ± 0.2 × 10-3 | 24 ± 6 | 1.1 ± 0.4 × 10-4 | 710 ± 70 |
H1242A | 9.8 × 10-1 | 20 | 1.7 ± 0.2 × 10-3 | 2300 ± 500 |
R1239A | 2.1 | 20 | 2.7 ± 0.3 × 10-3 | 2700 ± 500 |
H1242A/R1239A | 6.9 ± 0.5 × 10-2 | 37 ± 11 | 4.0 ± 0.4 × 10-4 | 2900 ± 500 |
M1238A | 5.6 ± 0.5 × 10-2 | 47 ± 17 | 3.6 ± 0.3 × 10-4 | 990 ± 230 |
- Incorporation of cGTP was studied in ECG1, misincorporation of ncGTP - in ECA. Kd and kpol (reaction rate at saturating nucleotide triphosphate concentration) for incorporation by R1239A and H1242A RNAPs were obtained by fitting the kinetics data into a simple kinetic model as described in Additional File 1: Supplementary Methods. Kd and kpol for incorporation by other RNAPs and misincorporation by all RNAPs was obtained by fitting into the Michaelis-Menten equation as described in Materials and Methods. Errors are the standard error of data fitting. The values were rounded to two significant digits.
