Table 4.
Alanine-scanning mutagenesis of proline residues in NS5A domain II
| Proline mutation (in DEYN) | Replication capacitya |
sh-A161/sh-Luc ratio of replication capacityb | |
|---|---|---|---|
| sh-Luc | sh-A161 | ||
| None/D316/Y317 | +++ | − | 0.04 ± 0.03 |
| None | ++++ | ++++ | 2.50 ± 0.32 |
| P310A | + | − | 0.05 ± 0.01 |
| P341-343A | ++ | − | 0.07 ± 0.06 |
| P310A/P341A | − | − | NA |
| P341A | ++++ | ++ | 0.24 ± 0.02 |
| P342A | +++ | ++++ | 1.94 ± 0.21 |
| P343A | +++ | ++++ | 2.71 ± 0.09 |
| P347A | + | ++++ | 4.28 ± 0.10 |
| P349-351A | ++++ | ++++ | 1.36 ± 0.08 |
| P379-380A | ++ | +++ | 1.49 ± 0.04 |
| P399A | +++ | ++++ | 2.85 ± 1.35 |
| P402A | ++++ | ++++ | 1.42 ± 0.38 |
| P405A | +++ | ++++ | 1.79 ± 0.50 |
| P407A | ++++ | ++++ | 1.08 ± 0.28 |
| P417-418A | ++++ | ++++ | 1.37 ± 0.75 |
| P423A | ++++ | ++++ | 1.11 ± 0.14 |
| P426A | ++++ | ++++ | 1.89 ± 0.09 |
| P437-439A | ++ | ++++ | 3.01 ± 1.20 |
| P446A | ++ | +++ | 2.92 ± 1.37 |
a
Replication capacity of the WT (fold increase from 4 h to day 3) was set at 100%, and the following scales were used: −, 0 to 10%; +, 11 to 50%; ++, 51 to 75%; +++, 76 to 100%; ++++, >100%.
b
The ratio of replication capacity in sh-A161 cells to that in sh-Luc cells was calculated and presented for all the D316E/Y317N mutation-based genomes. NA, not assayed.