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SOS induction by P1 Km miniplasmids.

M A Capage, J R Scott

    Journal of Bacteriology
    |August 1, 1983
    PubMed
    Summary

    P1 miniplasmids lacking EcoRI-6 triggered cellular SOS responses due to aberrant DNA replication initiation. The presence of EcoRI-6 partially restored normal replication, preventing some SOS pathway activations.

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    Area of Science:

    • Molecular Biology
    • Bacteriophage Genetics
    • DNA Replication

    Background:

    • Bacteriophage P1 is a model system for studying DNA replication and host-phage interactions.
    • Miniplasmids are essential tools for dissecting the minimal genetic elements required for replication.
    • The SOS response is a global regulatory network in bacteria, typically induced by DNA damage or replication stress.

    Purpose of the Study:

    • To construct and characterize functional P1 miniplasmids.
    • To investigate the role of specific P1 EcoRI fragments in replication and host-cell interactions.
    • To determine the cause of SOS pathway induction by P1 miniplasmids.

    Main Methods:

    • In vitro construction of P1 Km miniplasmids with varying EcoRI fragments.
    • Assays for SOS pathway induction: recA operon expression, e14 element excision, and lambda phage induction.
    • Analysis of miniplasmid replication origins and dependence on host factors (recA+).

    Main Results:

    • The smallest functional P1 miniplasmid replicon included EcoRI-5 (right side) and EcoRI-8.
    • Miniplasmids lacking EcoRI-6 induced SOS functions (recA expression, e14/lambda excision), linked to aberrant replication initiation.
    • EcoRI-6, in cis or trans, reduced SOS induction but still elevated recA expression, suggesting partial restoration of replication.

    Conclusions:

    • P1 EcoRI fragments 5 and 8 alone are insufficient for normal P1 replication, leading to SOS induction.
    • The P1 EcoRI-6 fragment, or its product, plays a role in ensuring proper replication initiation and mitigating SOS responses.
    • Aberrant initiation of DNA replication by P1 miniplasmids is a key trigger for cellular SOS functions.

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