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J. Biol. Chem., Vol. 268, Issue 32, 23762-23765, Nov, 1993
RM Schaaper
The accuracy by which organisms duplicate their DNA is of considerable
interest. At least three mechanisms operate, serially, to secure high
fidelity: base selection, exonucleolytic proofreading, and postreplicative
mismatch correction. To obtain insights into the efficiency and specificity
of these steps in the bacterium Escherichia coli, we have performed DNA
sequence analysis of mutations occurring in the bacterial lacI gene in a
series of strains genetically disabled in one or more of these error
avoidance pathways. The base selection efficiency was estimated from
mutagenesis occurring in a mutDmutL strain, which is deficient in both
proofreading (mutD5) and mismatch repair (mutL). The proofreading
efficiency was derived comparing the mutD5 mutL strain to the mismatch
repair-deficient mutL strain. The efficiency of mismatch repair was derived
comparing the mutL strain to the wild-type strain. The results show that
base selection discriminates against errors by 200,000-2,000,000-fold,
proofreading by 40-200-fold, and mismatch repair by 20-400-fold, each
depending on the type of error. Base selection and proofreading act more
strongly against transversions than transitions, whereas mismatch repair
does the opposite. The data are based on 866 sequenced lacI mutations in a
target that allows the scoring of at least 127 different mutations in 76
distinct DNA sequence contexts in vivo. They may therefore have general
significance.
Base selection, proofreading, and mismatch repair during DNA replication in Escherichia coli
Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709.
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