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Genetic exchanges induced by structural damage in nonreplicating phage lambda DNA.

P Howard-Flanders, P F Lin, E Bardwell

    Basic Life Sciences
    |January 1, 1975
    PubMed
    Summary
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    DNA damage from specific light treatments can induce genetic recombination in lambda phage. Photoproducts other than thymine dimers, and psoralen-induced cross-links processed by repair enzymes, significantly increase recombination rates.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Photochemistry

    Background:

    • Genetic recombination is a fundamental process in DNA repair and evolution.
    • Bacteriophage lambda serves as a model system for studying DNA damage and repair mechanisms.
    • Understanding DNA damage-induced recombination is crucial for fields ranging from cancer research to astrobiology.

    Purpose of the Study:

    • To investigate the role of specific DNA photoproducts in inducing genetic recombination in lambda phage.
    • To determine whether DNA replication is necessary for photoproducts to cause recombination.
    • To elucidate the mechanisms by which psoralen-induced cross-links and monoadducts lead to recombination.

    Main Methods:

    • Irradiation of lambda phage with different wavelengths and photosensitizers (254 nm, 313 nm with acetophenone D, 360 nm with trimethylpsoralen).

    Related Experiment Videos

  • Infection of homoimmune lysogens with treated phage under conditions blocking DNA replication and excision-repair.
  • Analysis of genetic recombination frequencies in two-factor crosses with varying marker separations.
  • Testing recombination in wild-type and mutant (uvrA, uvrB, polA) strains.

    • Main Results:

    • Thymine dimers alone, without DNA replication, did not significantly increase recombination.
    • 254 nm light, causing pyrimidine dimers and other photoproducts, markedly increased recombination in the absence of replication.
    • Psoralen (trimethylpsoralen) treatment significantly increased recombination in wild-type but not in uvrA/uvrB mutants.
    • Psoralen-induced cross-links appeared to act over distances of at least 5% of the genome, suggesting a role for excision enzymes and strand breaks.

    Conclusions:

    • Photoproducts other than thymine dimers can induce recombination independently of DNA replication.
    • Psoralen-induced recombination requires excision repair enzymes, potentially generating strand breaks that initiate recombination.
    • Cross-links can mediate recombination over significant genomic distances, possibly through heteroduplex formation.