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Plasmid pSC101 replication in integratively suppressed cells requires dnaA function.

J Felton, A Wright

    Molecular & General Genetics : MGG
    |September 1, 1979
    PubMed
    Summary
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    Plasmid pKC17 maintenance in E. coli depends on the dnaA gene. Suppressor mutations in rpoB allow plasmid stability at higher temperatures, even when the dnaA46 mutant is inactivated.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Plasmid maintenance is crucial for bacterial genetic stability.
    • The dnaA gene product is essential for DNA replication initiation in E. coli.
    • Plasmid pKC17, a pSC101 derivative, has specific maintenance requirements.

    Purpose of the Study:

    • To investigate the role of the dnaA gene product in plasmid pKC17 maintenance.
    • To determine the effect of temperature-sensitive dnaA mutations on plasmid stability.
    • To explore the impact of suppressor mutations on plasmid maintenance under stress conditions.

    Main Methods:

    • Segregation experiments were performed using an E. coli dnaA46 mutant.
    • Growth conditions involved various temperatures, with P2sig5 prophage enabling growth at elevated temperatures.

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  • The effect of rpoB mutations on plasmid pKC17 stability was assessed at different temperatures.

    • Main Results:

    • A functional dnaA gene product is required for the stable maintenance of plasmid pKC17.
    • The dnaA46 mutant exhibited temperature sensitivity, impacting plasmid stability at elevated temperatures.
    • rpoB suppressor mutations restored efficient maintenance of plasmid pKC17 up to 40°C, overcoming dnaA46 inactivation.

    Conclusions:

    • The dnaA gene product plays a critical role in the replication and stable inheritance of plasmid pKC17.
    • rpoB mutations can compensate for the loss of dnaA function regarding plasmid maintenance.
    • This study highlights the complex interplay between chromosomal and plasmid replication machinery in E. coli.