Systematic identification of synthetic lethal mutations with reduced-genome Escherichia coli : synthetic genetic interactions among yoaA, xthA and holC related to survival from MMS exposure

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  • Watanabe Keisuke
    Department of Biological Sciences, Graduate Schools of Science and Engineering, Tokyo Metropolitan University
  • Tominaga Kento
    Department of Biological Sciences, Graduate Schools of Science and Engineering, Tokyo Metropolitan University
  • Kitamura Maiko
    Department of Biological Sciences, Graduate Schools of Science and Engineering, Tokyo Metropolitan University
  • Kato Jun-ichi
    Department of Biological Sciences, Graduate Schools of Science and Engineering, Tokyo Metropolitan University

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  • Systematic identification of synthetic lethal mutations with reduced-genome <i>Escherichia coli</i>: synthetic genetic interactions among <i>yoaA</i>, <i>xthA</i> and <i>holC</i> related to survival from MMS exposure

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<p>Reduced-genome Escherichia coli strains lacking up to 38.9% of the parental chromosome have been constructed by combining large-scale chromosome deletion mutations. Functionally redundant genes involved in essential processes can be systematically identified using these reduced-genome strains. One large-scale chromosome deletion mutation could be introduced into the wild-type strain but not into the largest reduced-genome strain, suggesting a synthetic lethal interaction between genes removed by the deletion and those already absent in the reduced-genome strain. Thus, introduction of the deletion mutation into a series of reduced-genome mutants could allow the identification of other chromosome deletion mutations responsible for the synthetic lethal phenotype. We identified a synthetic lethality caused by disruption of nfo and xthA, two genes encoding apurinic/apyrimidinic (AP) endonucleases involved in the DNA base excision repair pathway, and two other large-scale chromosome deletions. We constructed temperature-sensitive mutants harboring quadruple-deletion mutations in the affected genes/chromosome regions. Using these mutants, we identified two multi-copy suppressors: holC, encoding the chi subunit of DNA polymerase III, and yoaA, encoding a putative DNA helicase. Addition of the yoaA disruption increased the methyl methanesulfonate (MMS) sensitivity of xthA single-deletion or xthA nfo double-deletion mutants. This increased MMS sensitivity was not suppressed by the presence of multi-copy holC. These results indicate that yoaA is involved in MMS sensitivity and suggest that YoaA functions together with HolC.</p>

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