Table 1.
Ad5 polymerase mutantsa
| Mutation | Type | Polymerase motif | Description of mutation | Result of mutation |
|---|---|---|---|---|
| I664V | C | (I/Y)xGG | Disrupts interaction of template DNA and polymerase | Viable |
| I664 M | C | (I/Y)xGG | Disrupts interaction of template DNA and polymerase | Viable (weakly) |
| R665K | C | (I/Y)xGG | Disrupts interaction of template DNA and polymerase | Viable (weakly) |
| M689V | C | A | Disrupts dNTP binding and substrate selection | Viable (weakly) |
| C687S | C | A | Disrupts dNTP binding and substrate selection | Viable (weakly) |
| I664S | NC | (I/Y)xGG | Disrupts interaction of template DNA and polymerase | Nonviable |
| I664Y | NC | (I/Y)xGG | Disrupts interaction of template DNA and polymerase | Nonviable |
| GG666/7AA | C | (I/Y)xGG | Disrupts interaction of template DNA and polymerase | Nonviable |
| M689I | C | A | Disrupts dNTP binding and substrate selection | Nonviable |
| G688S | C | A | Disrupts dNTP binding and substrate selection | Nonviable |
| Y690A | NC | A | Disrupts dNTP binding and substrate selection | Nonviable |
| Y690F | C | A | Disrupts dNTP binding and substrate selection | Nonviable |
| Y690I | NC | A | Disrupts dNTP binding and substrate selection | Nonviable |
| Y690V | NC | A | Disrupts dNTP binding and substrate selection | Nonviable |
| S692Y | NC | A | Disrupts dNTP binding and substrate selection | Nonviable |
| R833T | NC | B | Disrupts dNTP binding affinity | Nonviable |
| Y844S | NC | B | Disrupts dNTP binding affinity | Nonviable |
| K837N | NC | B | Disrupts dNTP binding affinity | Nonviable |
| N841E | NC | B | Disrupts dNTP binding affinity | Nonviable |
| N841Y | C | B | Disrupts dNTP binding affinity | Nonviable |
The table shows each mutant viral molecular clone that was constructed and analyzed. The position of each mutation is shown, along with the type of mutation (C, conservative amino acid substitution; NC, nonconservative substitution), the motif targeted ([I/Y]xGG, motif A, or motif B), the expected effect of the mutation, and the replication capacity of the viral molecular clone. Clones that replicated in Hek 293A host cells are indicated in boldface type. Clones were either nonviable (failed to replicate in any cell type, even with the addition of exogenous dNs), viable (replicated similarly to the wild-type virus in all cell types tested), or semiviable (replicated moderately well in Hek 293A cells but poorly in other cell types tested).