In search of a mutational hotspot.

Abstract:

In vitro selection was used to define sequence contexts that significantly enhanced the mutagenic potential of 7, 8-dihydro-8-oxoguanine (8-oxoG). Contexts that simultaneously reduced the efficiency of 8-oxoG cleavage by formamidopyrimidine DNA N-glycosylase and increased the efficiency of misincorporating A opposite the lesion by DNA polymerase were isolated from a pool of 4(8) random octanucleotide sequences. Kinetic analysis showed that the combined effects of poor repair and high miscoding resulted in 10(2)- to 10(3)-fold increase in the mutagenic potential of 8-oxoG. Furthermore, the isolated sequence contexts correlated strongly with G --> T transversion hotspots in spontaneous mutational spectra reported for the Escherichia coli lacI and human p53 and factor IX genes. We present an example directly linking the interplay between DNA repair and replication to a "high risk sequence" for base substitution.

Polymerases:

Topics:

Mutational Analysis, Modulators/Inhibitors, Kinetic Parameters, Nucleotide Analogs / Template Lesions, Fidelity, Accessory Proteins/Complexes, Nucleotide Incorporation

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
T4 D112AE114A Hatahet Z1998 KM 0.911uM Reaction: Nucleotide incorporation; Substrate: dCTP
T4 D112AE114A Hatahet Z1998 KM 0.32uM Reaction: Nucleotide incorporation; Substrate: dATP
T4 D112AE114A Hatahet Z1998 KM 1.01uM Reaction: Nucleotide incorporation; Substrate: dCTP
T4 D112AE114A Hatahet Z1998 KM 8.24uM Reaction: Nucleotide incorporation; Substrate: dATP
T4 D112AE114A Hatahet Z1998 KM 0.18uM Reaction: Nucleotide incorporation; Substrate: dATP
T4 D112AE114A Hatahet Z1998 KM 0.34uM Reaction: Misincorporation; Substrate: dATP
T4 D112AE114A Hatahet Z1998 KM 1.38uM Reaction: Misincorporation; Substrate: dATP
T4 D112AE114A Hatahet Z1998 KM 8.87uM Reaction: Misincorporation; Substrate: dATP
T4 D112AE114A Hatahet Z1998 KM 0.11uM Reaction: Misincorporation; Substrate: dCTP
T4 D112AE114A Hatahet Z1998 KM 0.332uM Reaction: Misincorporation; Substrate: dCTP
T4 D112AE114A Hatahet Z1998 KM 0.102uM Reaction: Nucleotide incorporation; Substrate: dATP
T4 D112AE114A Hatahet Z1998 KM 0.683uM Reaction: Misincorporation; Substrate: dCTP

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