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Related Experiment Videos

Correlation between UV dose requirement for lambda bacteriophage induction and lambda repressor concentration.

J Baluch, R Sussman

    Journal of Virology
    |June 1, 1978
    PubMed
    Summary

    UV radiation induces phage lambda in Escherichia coli. Higher lambda repressor levels decrease induction, while DNA repair mutations (uvrA) significantly lower the UV dose required for induction, impacting survival.

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    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • Bacteriophage lambda integrates into the Escherichia coli genome as a prophage.
    • The lambda cI repressor protein regulates the lysogenic cycle and can be inactivated by DNA damage.
    • DNA repair mechanisms, such as the uvrA-dependent pathway, influence cellular responses to UV irradiation.

    Purpose of the Study:

    • To investigate the impact of lambda repressor concentration and DNA repair capacity on phage lambda induction and survival following UV irradiation.
    • To compare the UV sensitivity of wild-type and uvrA mutant Escherichia coli lysogens with varying repressor levels.

    Main Methods:

    • Construction of isogenic Escherichia coli K-12 strains (wild-type and uvrA mutant) with multiple lambda cI genomes.
    • Quantification of lambda repressor concentration using in vitro DNA-binding assays.

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  • Determination of phage induction and cell survival curves after UV irradiation.

    • Main Results:

    • Both wild-type and uvrA mutant lysogens exhibited dose-dependent responses to UV irradiation, influenced by repressor concentration.
    • uvrA lysogens required significantly lower UV fluences (one-tenth) for phage induction compared to wild-type.
    • Overproduction of functional lambda repressor (lambdaind+) in uvrA lysogens led to infectious center formation upon UV exposure, unlike non-functional repressor (lambdaind-).

    Conclusions:

    • Cellular repressor concentration and DNA repair proficiency are critical determinants of UV-induced phage lambda induction and lysogen survival.
    • The uvrA mutation confers increased sensitivity to UV-induced prophage induction.
    • Functional lambda repressor is essential for the production of infectious phage particles following UV irradiation in DNA repair-deficient lysogens.