TABLE 2.
Rates of FRET phases
The phase rates (s−1) and errors at 20 °C were obtained by fitting stopped-flow fluorescence traces with the KinTek Explorer global fitting software (see “Experimental Procedures”) (20).
| Domain mutanta and DNA substrateb | Phase rate |
|||||
|---|---|---|---|---|---|---|
| Donor |
Acceptor |
|||||
| P0 | P1 | P2 | P0 | P1 | P2 | |
| s−1 | ||||||
| Finger N70C | ||||||
| O-2 | 15 ± 2 | 0.83 ± 0.04 | 14 ± 2 | 0.63 ± 0.06 | ||
| O-3 | 12 ± 1 | 18 ± 2 | ||||
| O-4 | 11.1 ± 0.8 | 0.38 ± 0.02 | 6 ± 2 | 0.41 ± 0.05 | ||
| O-5 | 9 ± 3 | 11 ± 2 | ||||
| Finger E49C | ||||||
| O-2 | 22 ± 4 | 0.41 ± 0.02 | 18 ± 5 | 0.49 ± 0.06 | ||
| O-3 | 22 ± 4 | 17 ± 3 | ||||
| Palm S112C | ||||||
| O-2 | >100 | 12 ± 0.8 | 0.4 ± 0.1 | >100 | 14 ± 1 | 0.17 ± 0.04 |
| O-3 | >100 | 12 ± 1 | >100 | 11 ± 2 | ||
| O-4 | 21 ± 2 | 1.33 ± 0.07 | 0.19 ± 0.01 | 13 ± 7 | 1.8 ± 0.5 | 0.19 ± 0.07 |
| O-5 | 21.8 ± 0.3 | 1.42 ± 0.02 | 21 ± 1 | 2.0 ± 0.1 | ||
| Thumb S207C | ||||||
| O-2 | 5.5 ± 0.8 | 0.24 ± 0.06 | 5.5 ± 0.7 | 0.12 ± 0.04 | ||
| O-3 | 17 ± 3 | 12 ± 2 | ||||
| O-4 | 6.4 ± 0.4 | 9 ± 1 | ||||
| O-5 | 8 ± 2 | 8.0 ± 0.5 | ||||
| LF K329C | ||||||
| O-2 | 9 ± 1 | 0.32 ± 0.03 | 5 ± 2 | 0.23 ± 0.07 | ||
| O-3 | 6.1 ± 0.5 | 5.4 ± 0.4 | ||||
| O-4 | 8.3 ± 0.3 | 0.51 ± 0.05 | 7.4 ± 0.6 | 0.30 ± 0.01 | ||
| O-5 | 5 ± 1 | 5.2 ± 0.4 | ||||
a Each of the Alexa Fluor 594-labeled Dpo4 mutants also contains the C31S mutation and is listed in supplemental Table S1.
b DNA substrates are listed in Table 1.