Journal
MOLECULAR MICROBIOLOGY
Volume 86, Issue 6, Pages 1364-1375Publisher
WILEY-BLACKWELL
DOI: 10.1111/mmi.12061
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Funding
- Ministry of Education, Culture, Sports, Science and Technology, Japan (MEXT) [18201010, 22241016]
- Ministry of Health, Labour and Welfare, Japan (MHLW) [H21-Food-General-009]
- Japan Health Science Foundation [KHB1007]
- MHLW [20 designated-8]
- Food Safety Commission
- Grants-in-Aid for Scientific Research [18201010, 22241016] Funding Source: KAKEN
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Reactive oxygen species induce oxidative damage in DNA precursors, i.e. dNTPs, leading to point mutations upon incorporation. Escherichia coli mutT strains, deficient in the activity hydrolysing 8-oxo-7,8-dihydro-2'-deoxyguanosine 5'-triphosphate (8-oxo-dGTP), display more than a 100-fold higher spontaneous mutation frequency over the wild-type strain. 8-oxo-dGTP induces A to C transversions when misincorporated opposite template A. Here, we report that DNA pol III incorporates 8-oxo-dGTP approximate to 20 times more efficiently opposite template A compared with template C. Single, double or triple deletions of pol I, pol II, pol IV or pol V had modest effects on the mutT mutator phenotype. Only the deletion of all four polymerases led to a 70% reduction of the mutator phenotype. While pol III may account for nearly all 8-oxo-dGTP incorporation opposite template A, it only extends approximate to 30% of them, the remaining 70% being extended by the combined action of pol I, pol II, pol IV or pol V. The unique property of pol III, a C-family DNA polymerase present only in eubacteria, to preferentially incorporate 8-oxo-dGTP opposite template A during replication might explain the high spontaneous mutation frequency in E.?coli mutT compared with the mammalian counterparts lacking the 8-oxo-dGTP hydrolysing activities.
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