TABLE 4.
Kinetic parameters calculated from the RNA unwinding (Uw) and rewinding (Rw) experiments
P5′Uw and P15′Rw, percentage of ssRNA and dsRNA formed at 5 and 15 min, respectively; AUw and ARw, final reaction amplitude.
| Types of substrate | Unwinding |
Rewindinga |
||||||
|---|---|---|---|---|---|---|---|---|
| NsP2 |
His-nsP2 |
NsP2 |
His-nsP2 |
|||||
| P5′Uw | AUw | P5′Uw | AUw | P15′Rw | ARw | P15′Rw | ARw | |
| % of ssRNA formed | % of dsRNA formed | |||||||
| 12-Base 5′ overhang | 24.3 ± 5.0 | 48.0 ± 6.7 | 14.2 ± 5.4 | 24.6 ± 4.3 | 3.5 ± 1.8 | 9.8 ± 0.4 | 23.9 ± 0.4 | 45.4 ± 1.0 |
| 24-Base 5′ overhang | 34.7 ± 1.7 | 54.3 ± 1.7 | 33.9 ± 0.3 | 29.9 ± 2.3b | 11.4 ± 1.9 | 16.7 ± 4.3 | 30.4 ± 2.5 | 39.5 ± 1.5 |
| 12-Base 3′ and 5′ overhangs (fork) | 20.1 ± 0.8 | 36.1 ± 6.1 | 11.3 ± 0.1 | 28.5 ± 4.3 | 30.0 ± 0.2 | 47.1 ± 0.7 | 47.4 ± 3.6 | 64.3 ± 0.1 |
a The activities represent the difference between the amount of dsRNA formed in the presence and absence of the enzyme.
b In this reaction, the maximal reaction amplitude was 46.5 ± 1.9% (Fig. 5A).