Effect of temperature on transition and transversion mutagenesis: characterization of wild type and a mutator T4 DNA polymerase mutant.

Abstract:

The effect of temperature on genetically well-defined mutational pathways was examined in the bacteriophage T4. The mutational site was a T4 rII ochre mutant which could revert to rII+ via a transversion or to the amber convertant via a transition. Temperature did not strongly affect any of the pathways examined in a wild-type background; however, increased temperature reduced the mutational activity of a mutator DNA polymerase mutant. Possible models to explain the role of temperature in mutagenesis are discussed as well as the significance of low temperatures for in vitro mutagenesis reactions.

Polymerases:

Topics:

Mutational Analysis, Fidelity, Nucleotide Incorporation, Exonuclease Activity

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
T4 L340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 2000 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->GC pathway (P7oc -> P7am), 30°C)
T4 L340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 333 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->TA(CG) pathway (P7oc -> rII+), 30°C)
T4 L340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 750 Mutation frequency (relative to WT) Technique: T4 rII P7am reversion assay (GC->TA(CG) pathway (P7am -> rII+), 30°C)
T4 V325AL340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 34 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->GC pathway (P7oc -> P7am), 43°C)
T4 V325AL340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 36 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->TA(CG) pathway (P7oc -> rII+), 43°C)
T4 V325AL340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 20 Mutation frequency (relative to WT) Technique: T4 rII P7am reversion assay (GC->TA(CG) pathway (P7am -> rII+), 43°C)
T4 V325AL340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 480 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->GC pathway (P7oc -> P7am), 30°C)
T4 V325AL340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 2500 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->TA(CG) pathway (P7oc -> rII+), 30°C)
T4 V325AL340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 500 Mutation frequency (relative to WT) Technique: T4 rII P7am reversion assay (GC->TA(CG) pathway (P7am -> rII+), 30°C)
T4 V325AL340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 214 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->GC pathway (P7oc -> P7am), 37°C)
T4 V325AL340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 333 Mutation frequency (relative to WT) Technique: T4 rII P7oc reversion assay (AT->TA(CG) pathway (P7oc -> rII+), 37°C)
T4 V325AL340P Reha-Krantz LJ1984 Nucleotide Substitution Rate 36 Mutation frequency (relative to WT) Technique: T4 rII P7am reversion assay (GC->TA(CG) pathway (P7am -> rII+), 37°C)

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