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

Mutation research (1996), Volume 350, Page 9

PubMed entry

Abstract:

During DNA replication, DNA polymerases alternate between DNA ...

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:

T4 D863up/R335C,T4 D863up/S345F,T4 S345F,T4 R335C,T4,T4 D863 uplication

Topics:

Mutational Analysis, Fidelity, Methods

Status:

Results:

Polymerase	Reference	Property	Result	Context
T4 D863up/S345F	Selection of bacteriophage T4 antimutator DNA polymerases: a link between proofreading and sensitivity to phosphonoacetic acid.	Nucleotide Substitution Rate	0.05 Mutation frequency (relative to WT)	Technique: T4 rII 199oc reversion assay (30°C)
T4 S345F	Selection of bacteriophage T4 antimutator DNA polymerases: a link between proofreading and sensitivity to phosphonoacetic acid.	Nucleotide Substitution Rate	0.02 Mutation frequency (relative to WT)	Technique: T4 rII 199oc reversion assay (30°C)
T4 D863up/R335C	Selection of bacteriophage T4 antimutator DNA polymerases: a link between proofreading and sensitivity to phosphonoacetic acid.	Nucleotide Substitution Rate	0.1 Mutation frequency (relative to WT)	Technique: T4 rII 199oc reversion assay (30°C)
T4 R335C	Selection of bacteriophage T4 antimutator DNA polymerases: a link between proofreading and sensitivity to phosphonoacetic acid.	Nucleotide Substitution Rate	0.01 Mutation frequency (relative to WT)	Technique: T4 rII 199oc reversion assay (30°C)
T4 D863 uplication	Selection of bacteriophage T4 antimutator DNA polymerases: a link between proofreading and sensitivity to phosphonoacetic acid.	Nucleotide Substitution Rate	10 Mutation frequency (relative to WT)	Technique: T4 rII 199oc reversion assay (30°C)