Selection of bacteriophage T4 antimutator DNA polymerases: a link between proofreading and sensitivity to phosphonoacetic acid.

Mutation research (1996), Volume 350, Page 9

Abstract:

During DNA replication, DNA polymerases alternate between DNA synthesis and proofreading the newly synthesized DNA. In order to understand the molecular details of how DNA polymerases determine the balance between polymerase and proofreading activities, it would be useful to have mutants which switch between the two activities either more or less frequently. Antimutator DNA polymerases switch more frequently and thus have more opportunity for proofreading. We have observed that mutant DNA polymerases which proofread less frequently have a mutator phenotype and are inhibited by the pyrophosphate analogue phosphonoacetic acid. Sensitivity to phosphonoacetic acid can be used to isolate second-site suppressor mutations. These suppressor mutations encode amino acid substitutions which produce antimutator DNA polymerases.

Polymerases:

Topics:

Mutational Analysis, Fidelity, Methods

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
T4 D863up/S345F Reha-Krantz LJ1996 Nucleotide Substitution Rate 0.05 Mutation frequency (relative to WT) Technique: T4 rII 199oc reversion assay (30°C)
T4 S345F Reha-Krantz LJ1996 Nucleotide Substitution Rate 0.02 Mutation frequency (relative to WT) Technique: T4 rII 199oc reversion assay (30°C)
T4 D863up/R335C Reha-Krantz LJ1996 Nucleotide Substitution Rate 0.1 Mutation frequency (relative to WT) Technique: T4 rII 199oc reversion assay (30°C)
T4 R335C Reha-Krantz LJ1996 Nucleotide Substitution Rate 0.01 Mutation frequency (relative to WT) Technique: T4 rII 199oc reversion assay (30°C)
T4 D863 uplication Reha-Krantz LJ1996 Nucleotide Substitution Rate 10 Mutation frequency (relative to WT) Technique: T4 rII 199oc reversion assay (30°C)

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