TABLE 2.
In vivo mutation rates and in vitro elongation data
| 3Dpol | Location | No. of mutations/10 kba | Max elongation rate (nt/s) ± SEb | CTP Km (nM) ± SEc | CTP-vs-dCTP discrimination factor ± SEc |
|---|---|---|---|---|---|
| Coxsackievirus B3 | |||||
| Wild type | 4.3 | 22.8 ± 1.5 | 115 ± 7 | 360 ± 40 | |
| I230F | Palm/motif A | 11.2 | 30.4 ± 1.0 | 116 ± 7 | 230 ± 30 |
| I230W | Palm/motif A | NV | 26 ± 4d | 93 ± 7 | 330 ± 40 |
| I230L | Palm/motif A | ND | 34 ± 5d | 88 ± 7 | 240 ± 30 |
| F232Y | Palm/motif A | 11.2 | 28 ± 5 | ND | 270 ± 50e |
| F232L | Palm/motif A | Rev | 34.6 ± 1.5 | 128 ± 5 | 174 ± 8 |
| F232V | Palm/motif A | Rev | 29.9 ± 1.4 | ND | 240 ± 40e |
| S299T | Palm/motif B | 7.0 | 19.7 ± 0.7 | 92 ± 6 | 290 ± 30 |
| K360R | Palm/motif D | NV | 0.6 ± 0.1 | ND | ND |
| G64S | Palm/N terminus | IVT | 12.6 ± 0.5 (37°C) | ND | 420 ± 70 (30°C) |
| A239G | Palm/N terminus | 8.2 | 37 ± 3 | 116 ± 7 | 169 ± 12 |
| Y268W | Fingers-palm | 8.9 | 25 ± 3 | 101 ± 5 | 310 ± 30 |
| Y268H | Fingers-palm | 9.2 | 28 ± 2 | 103 ± 10 | 280 ± 40 |
| P48K | Fingers | 6.8 | 28.1 ± 1.0 | 79 ± 3 | 320 ± 60 |
| S164P | Fingers | 6.7 | 21 ± 3 | 91 ± 3 | 340 ± 18 |
| I176V | Fingers-RNA | 5.6 | 30 ± 2 | 89 ± 8 | 310 ± 30 |
| CV double mutants | |||||
| I230F + S299T | Motif A + palm | 7.2 | 25.7 ± 1.6 | 106 ± 7 | 255 ± 20 |
| I230F + F232Y | Motif A + motif A | ND | 37 ± 5d | 114 ± 10 | 190 ± 20 |
| I230F + I176V | Motif A + fingers | ND | 38 ± 3 | 77 ± 3 | 234 ± 17 |
| G64S + I176V | N terminus + fingers | Rev | 17.2 ± 1.0 | ND | ND |
| G64S + Y268H | N terminus + fingers-palm | NV | 14.6 ± 1.9 (37°C) | ND | 350 ± 50 (30°C) |
| Poliovirus | |||||
| Wild type | 88 ± 5 | 85 ± 7 | 117 ± 11 | ||
| G64S | Palm/N terminus | ND | 46 ± 2 | 66 ± 5 | 310 ± 30 |
| F230I | Palm/motif A | ND | 75 ± 8 | 73 ± 3 | 152 ± 9 |
| F232I | Palm/motif A | ND | 56 ± 16 | 92 ± 2 | 200 ± 13 |
| H273R | Fingers | ND | 86 ± 14 | 123 ± 7 | 57 ± 19 |
| K359R | Palm/motif D | ND | 3.0 ± 0.1 | 34 ± 4 | 1,300 ± 500 |
From sequencing of progeny virus genomes as described previously by Gnadig et al. (19). NV, virus was not viable; IVT, insufficient virus titer for mutational analysis; ND, not determined; Rev, virus was viable but reverted to the wild-type residue.
Maximal rate from Michaelis-Menten analysis of the lag phase in 37°C all-NTP stopped-flow assays.
From Michaelis-Menten analysis of CTP and 2′-dCTP titration data with propagation of fractional errors in the curve-fitted parameters (see Fig. 4C and Table 3 for further details regarding 30°C data).
Obtained from the length of the lag phase in the discrimination assay.
Scaled from data obtained with the original RNA template used by Gnadig et al. (19).