TABLE 2.
Summary of mutagenesis results
| Replicon | Translation (%) | Replication (%)a | Reversionb |
|---|---|---|---|
| 1st bp | |||
| 79A-U1 (WT) | 100 | 100 | |
| 79AxC1 | 98 | 15 | Partial (C or U) |
| 79AxA1 | 106 | 16 | Yes |
| 79AxG1 | 96 | 14 | Yes |
| 79UxU1 | 113 | <1 | ND |
| 79U-A1 | 109 | <1 | ND |
| 79G-U1 | 99 | 132 | No |
| 2nd bp | |||
| 78G-C2 (WT) | 100 | 100 | |
| 78G-U2 | 102 | <1 | ND |
| 78GxA2 | 101 | <1 | ND |
| 78GxG2 | 115 | <1 | ND |
| 78CxC2 | 113 | <1 | ND |
| 3rd bp | |||
| 77A-U3 (WT) | 100 | 100 | |
| 77AxC3 | 108 | <1 | ND |
| 77AxA3 | 107 | <1 | ND |
| 77AxG3 | 105 | <1 | ND |
| 77UxU3 | 97 | <1 | ND |
| 77G-U3 | 96 | 23 | No |
| 77U-A3 | 98 | 56 | No |
| 4th bp | |||
| 76U-A4 (WT) | 100 | 100 | |
| 76UxU4 | 120 | <1 | ND |
| 76U-C4 | 107 | <1 | ND |
| 76U-G4 | 104 | 6 | No |
| 5th bp | |||
| 75C-G5 (WT) | 100 | 100 | |
| 75CxA5 | 92 | <1 | ND |
| 75CxC5 | 112 | <1 | ND |
| 75CxU5 | 110 | <1 | ND |
| 75GxG5 | 84 | <1 | ND |
| 75AxG5 | 83 | <1 | ND |
| 75U-G5 | 90 | 3.5 | No |
| 75G-C5 | 97 | <1 | ND |
| 6th bp | |||
| 74U-G6 (WT) | 100 | 100 | |
| 74UxC6 | 101 | 27 | No |
| 74UxU6 | 111 | 73 | No |
| 74U-A6 | 112 | 181 | No |
a
The translation and replication levels of mutant replicons are expressed as the replicons' Rluc activities as a percentage of the wild-type (WT) level at 2.5 and 72 h posttransfection, respectively.
b
For reversion analysis, mutant replicons were extracted at 72 h posttransfection, polyadenylated, and subjected to 3′ RACE analysis (see details in Materials and Methods). ND, not determined.