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Related Experiment Videos

[Mutagenic action of nitrous acid on prophage lambda].

S E Bresler, V L Kalinin, L V Kuznetsova

    Genetika
    |January 1, 1982
    PubMed
    Summary

    Nitrous acid causes lethal and mutagenic effects on lambda prophage in Escherichia coli. Delayed induction significantly increases prophage survival and alters mutation frequency, highlighting the role of DNA repair mechanisms.

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    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • Nitrous acid is a known mutagen.
    • Bacteriophage lambda and its prophage state in Escherichia coli serve as a model system for studying DNA damage and repair.
    • DNA repair pathways are crucial for cellular survival and genomic stability.

    Purpose of the Study:

    • To investigate the lethal and mutagenic effects of nitrous acid on lambda prophage in wild-type E. coli and various repair-deficient mutants.
    • To determine the influence of induction timing (immediate vs. delayed) on prophage survival and mutation frequency.
    • To elucidate the roles of specific DNA repair genes (e.g., recA, lexA, uvr) in mediating the response to nitrous acid treatment.

    Main Methods:

    • Treatment of E. coli lysogenic for lambda prophage with nitrous acid (HNO2).
    • Heat induction of prophage, performed either immediately after treatment or after a 90-minute incubation period.
    • Quantification of prophage survival and the frequency of clear plaque mutants (c mutants) in wild-type and repair-deficient strains.

    Main Results:

    • Delayed prophage induction significantly increased survival compared to immediate induction.
    • Lethal effects of nitrous acid were more pronounced in uvrA and uvrE mutants after delayed induction.
    • Mutation frequency was substantially higher in several repair-deficient mutants (uvrA, uvrA umuC, uvrD, uvrE, polA, xthA) after immediate induction.
    • RecA and LexA proteins were essential for most mutations under immediate induction, suggesting error-prone repair (W-mutagenesis).
    • Under delayed induction, mutation frequency decreased significantly in wild-type and several mutant strains, with mutations appearing less dependent on RecA and LexA, possibly fixed by replication.

    Conclusions:

    • The timing of prophage induction critically affects the outcome of nitrous acid mutagenesis.
    • DNA repair pathways, particularly those involving RecA and LexA, play a major role in generating mutations after nitrous acid treatment under immediate induction.
    • Under delayed induction, replication appears to be the primary mechanism for fixing nitrous acid-induced mutations, with reduced reliance on RecA/LexA-dependent pathways.

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