TABLE 3.
The DNA polymerase assay with 5 mM ATP and with modification of dNTP concentration or template sequence
| Mutations in wild type background | Assay modifications of dNTP concn or template sequence | NRTI-TPa
|
|||||
|---|---|---|---|---|---|---|---|
| DOT-TP
|
TFV-DP
|
AZT-TP
|
|||||
| IC50 ± SE | Fold increase | IC50 ± SE | Fold increase | IC50 ± SE | Fold increase | ||
| Wild-type HXB2(a) | 1 μM dNTP | 0.38 ± 0.04 | 1.0 | 3.3 ± 0.2 | 1.0 | 0.46 ± 0.05 | 1.0 |
| Wild-type HXB2(a) | 20 μM dNTP | 2.0 ± 0.1 | 1.0 | 19 ± 0.5 | 1.0 | 3.3 ± 0.7 | 1.0 |
| 41L/67N/70R/210W/215Y | 1 μM dNTP | 0.5 ± 0.2 | 1.3 | 13 ± 2b | 3.7 | 2.3 ± 0.7b | 5.0 |
| 41L/67N/70R/210W/215Y | 20 μM dNTP | 2.4 ± 0.3 | 1.2 | 49 ± 7b | 2.6 | 16 ± 3b | 4.8 |
| Wild-type HXB2(a) | 5′-A12-(GTCA)5-3′ | 0.38 ± 0.04 | 1.0 | 3.3 ± 0.2 | 1.0 | 0.46 ± 0.05 | 1.0 |
| Wild-type HXB2(a) | 5′-A12-(GACT)5-3′ | 0.31 ± 0.01 | 1.0 | 2.8 ± 0.3 | 1.0 | ND | ND |
| 41L/67N/70R/210W/215Y | 5′-A12-(GTCA)5-3′ | 0.5 ± 0.2 | 1.3 | 13 ± 2b | 3.7 | 2.3 ± 0.7b | 5.0 |
| 41L/67N/70R/210W/215Y | 5′-A12-(GACT)5-3′ | 0.4 ± 0.03b | 1.3 | 8.6 ± 1b | 3.1 | ND | ND |
| 41L/69S-SG/210W/215Y | 5′-A12-(GTCA)5-3′ | 1.6 ± 0.2b | 4.2 | 24 ± 5b | 7.3 | 4.4 ± 1.2b | 9.6 |
| 41L/69S-SG/210W/215Y | 5′-A12-(GACT)5-3′ | 0.78 ± 0.1b | 2.5 | 17 ± 1b | 6.0 | ND | ND |
a
Mean IC50 values ± standard errors (± SE) are expressed as μM from at least two separate experiments conducted in duplicate. ND, not determined. Fold increase was calculated by dividing the mutant RT IC50 values by respective wild-type IC50 values and assay modification.
b
Differs statistically from matched wild-type IC50 (P < 0.05) values.