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Recombination of uracil-containing lambda bacteriophages.

J B Hays, B K Duncan, S Boehmer

    Journal of Bacteriology
    |January 1, 1981
    PubMed
    Summary
    This summary is machine-generated.

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    Controlled uracil incorporation into bacteriophage DNA increases recombination frequencies. Uracil residues are lethal to phages but stimulate recombination, with excision repair playing a key role.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Deoxyribonucleic acid (DNA) normally contains thymine, but uracil can be incorporated under specific conditions.
    • The presence of uracil in DNA can impact DNA replication, repair, and recombination processes.
    • Bacteriophages, such as lambda phage, are valuable model systems for studying DNA metabolism.

    Purpose of the Study:

    • To investigate the controlled incorporation of uracil into lambda bacteriophage DNA.
    • To determine the biological consequences of uracil substitution for thymine on phage viability and recombination.
    • To elucidate the role of DNA repair pathways in processing uracil-containing DNA.

    Main Methods:

    • Bacterial mutants (dut ung thy) of Escherichia coli were used to grow lambda bacteriophages with incorporated uracil.

    Related Experiment Videos

  • Alkaline sucrose sedimentation and uracil DNA glycosylase treatment were employed to quantify uracil incorporation.
  • Phage plating efficiency assays on wild-type and mutant bacteria assessed uracil's impact on viability.
  • Recombination frequencies were measured using lambda tandem duplication phages in repressed infections, with genetic analysis of recombination-deficient mutants (recA, recB, xth).

    • Main Results:

    • Uracil substitution for thymine in phage DNA ranged from 0.17% to 1.9%.
    • Uracil incorporation reduced phage plating efficiency, with a lethality probability of approximately 1% per uracil residue.
    • Uracil-containing phages exhibited 5 to 10 times higher recombination frequencies compared to controls.
    • Uracil-stimulated recombination was dependent on RecA and RecB proteins and significantly enhanced in xth mutants, suggesting a role for excision repair.

    Conclusions:

    • Controlled uracil incorporation into bacteriophage DNA is feasible and impacts phage biology.
    • Uracil acts as a mutagenic lesion, reducing viability but also stimulating recombination.
    • Excision repair pathways are crucial for processing uracil in DNA and influence uracil-mediated recombination.
    • This study provides insights into DNA repair mechanisms and the consequences of base modification in viral genomes.