Mutagenesis induced by oxidized DNA precursors: roles of Y family DNA polymerases in Escherichia coli.

Abstract:

To reveal the roles of Y family DNA polymerases in the mutagenesis induced by oxidatively damaged DNA precursors, 2-hydroxy-dATP (2-OH-dATP) and 8-hydroxy-dGTP (8-OH-dGTP) were introduced into Escherichia coli strains deficient in the Y family polymerases, DNA polymerase IV (pol IV, encoded by the dinB gene) and DNA polymerase V (pol V, encoded by the umuDC locus). The mutation induced by 2-OH-dATP, but not that induced by 8-OH-dGTP, occurred less frequently in the dinB- strain than in the wild-type (wt) strain, suggesting the involvement of pol IV in the mutagenesis by 2-OH-dATP. Expression of pol IV from plasmid enhanced the mutagenesis by 2-OH-dATP in the dinB- strain. This enhancement depends on the polymerase activity since the expression of a mutant pol IV lacking the polymerase activity did not increase the mutations induced by 2-OH-dATP. In contrast, both 2-OH-dATP and 8-OH-dGTP caused mutations more efficiently in the umuDC- strain than in the wt strain, suggesting that the umuDC gene products suppressed the mutagenesis by these oxidized DNA precursors. The DNA polymerase activity was not required for the suppressive effects because expression of the umuDC gene products lacking the polymerase activity also suppressed the mutagenesis. These results suggest that the E. coli pol IV was involved in mutagenesis by 2-OH-dATP and that the umuDC gene products play suppressive role(s) in the mutagenesis by damaged nucleotides.

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