Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant.
Abstract:
We describe here our first attempt in using suppressor mutations to study structure-function relationships of the bacteriophage T4 DNA polymerase. One intragenic suppressor mutation, J5(43) degrees, was isolated that suppresses the temperature sensitivity but not the mutator activity of tsM19, a DNA polymerase mutant. Thus, the substituted amino acid induced by the tsM19 lesion decreases DNA polymerase fidelity, even if the temperature sensitivity has been corrected by a second amino acid substitution in the DNA polymerase polypeptide. The isolation, mapping and characterization of the J5(43) degrees mutation as well as the purification and characterization of the tsM19-J5(43) degrees mutant DNA polymerase are presented. The suppressor isolation procedure has general applicability for the selection of suppressor mutations of other T4 DNA polymerase mutator mutants.
Polymerases:
Topics:
Mutational Analysis, Structure and Structure/Function, Fidelity, Nucleotide Incorporation, Methods
Status:
| new | topics/pols set | partial results | complete | validated |
Results:
| Polymerase | Reference | Property | Result | Context |
|---|---|---|---|---|
| T4 Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 0.6 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->GC, Host: supE (C600)) |
| T4 Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 0.2 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->GC, Host: supD (CR63)) |
| T4 Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 1.2 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supE (C600)) |
| T4 Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 2.2 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supE (C600)) |
| T4 Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 0.1 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supD (CR63)) |
| T4 Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 0.4 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supD (CR63)) |
| T4 L340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 238 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supD (CR63)) |
| T4 L340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 538 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supD (CR63)) |
| T4 L340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 1189 Mutation frequency (relative to WT) | Technique: ac forward mutational assay (Host: supE (MBM7007.2)) |
| T4 L340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 1744 Mutation frequency (relative to WT) | Technique: ac forward mutational assay (Host: supD (MBM7007.1)) |
| T4 L340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 1980 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->GC, Host: supE (C600)) |
| T4 L340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 343 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supE (C600)) |
| T4 L340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 742 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supE (C600)) |
| T4 L340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 941 Mutation frequency (relative to WT) | Technique: T4 rII 199oc reversion assay (AT->GC, Host: supD (CR63)) |
| T4 V325AL340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 844 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supD (CR63)) |
| T4 V325AL340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 674 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supD (CR63)) |
| T4 V325AL340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 2305 Mutation frequency (relative to WT) | Technique: ac forward mutational assay (Host: supE (MBM7007.2)) |
| T4 V325AL340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 4160 Mutation frequency (relative to WT) | Technique: ac forward mutational assay (Host: supD (MBM7007.1)) |
| T4 V325AL340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 480 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay ( AT->GC, Host: supE (C600)) |
| T4 V325AL340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 2500 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supE (C600)) |
| T4 V325AL340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 500 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supE (C600)) |
| T4 V325AL340P | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 642 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay ( AT->GC, Host: supD (CR63)) |
| T4 L340PQ731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 6 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supD (CR63)) |
| T4 L340PQ731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 4 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supD (CR63)) |
| T4 L340PQ731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 2930 Mutation frequency (relative to WT) | Technique: ac forward mutational assay (Host: supE (MBM7007.2)) |
| T4 L340PQ731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 96 Mutation frequency (relative to WT) | Technique: ac forward mutational assay (Host: supD (MBM7007.1)) |
| T4 L340PQ731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 1720 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->GC, Host: supE (C600)) |
| T4 L340PQ731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 1417 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supE (C600)) |
| T4 L340PQ731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 1600 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supE (C600)) |
| T4 L340PQ731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 20 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->GC, Host: supD (CR63)) |
| T4 V325AL340P/Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 35 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supD (CR63)) |
| T4 V325AL340P/Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 17 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supD (CR63)) |
| T4 V325AL340P/Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 4453 Mutation frequency (relative to WT) | Technique: ac forward mutational assay (Host: supE (MBM7007.2)) |
| T4 V325AL340P/Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 223 Mutation frequency (relative to WT) | Technique: ac forward mutational assay (Host: supD (MBM7007.1)) |
| T4 V325AL340P/Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 590 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->GC, Host: supE (C600)) |
| T4 V325AL340P/Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 1967 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay (AT->TA(CG), Host: supE (C600)) |
| T4 V325AL340P/Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 1867 Mutation frequency (relative to WT) | Technique: T4 rII P7am reversion assay (GC->TA(CG), Host: supE (C600)) |
| T4 V325AL340P/Q731am | Structure-function studies of the bacteriophage T4 DNA polymerase. Isolation of a novel suppressor mutant. | Nucleotide Substitution Rate | 49 Mutation frequency (relative to WT) | Technique: T4 rII P7oc reversion assay ( AT->GC, Host: supD (CR63)) |