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Postreplication repair in Saccharomyces cerevisiae

M A Resnick, J Boyce, B Cox

    Journal of Bacteriology
    |April 1, 1981
    PubMed
    Summary
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    Postreplication repair in ultraviolet-irradiated yeast mutants does not involve recombination. DNA interruptions are repaired without significant pyrimidine dimer exchange, suggesting alternative repair pathways.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • DNA damage from ultraviolet (UV) radiation poses a significant threat to cell survival.
    • Excision repair is a primary mechanism for removing UV-induced DNA lesions.
    • Understanding postreplication repair (PRR) is crucial for comprehending cellular responses to DNA damage, especially in mutants deficient in excision repair.

    Purpose of the Study:

    • To investigate the mechanisms of postreplication repair in Saccharomyces cerevisiae mutants lacking excision repair capabilities.
    • To determine if molecular recombination is involved in the repair of DNA gaps formed during replication post-UV irradiation.
    • To characterize the fate of pyrimidine dimers during postreplication repair in these mutants.

    Main Methods:

    • Utilizing pulse-labeling and chase experiments to track newly synthesized DNA in UV-irradiated excision-defective yeast mutants.

    Related Experiment Videos

  • Employing alkaline sucrose gradient sedimentation to analyze the size of chromosomal DNA.
  • Assaying pyrimidine dimers using UV-endonuclease activity from Micrococcus luteus extracts.

    • Main Results:

    • Pulse-labeled DNA in UV-irradiated mutants exhibited transient interruptions.
    • These DNA interruptions were resolved during the chase period, indicating repair.
    • The resolution of DNA interruptions occurred without significant exchange of pyrimidine dimers, with less than 10-20% of induced dimers being exchanged.

    Conclusions:

    • Postreplication repair in excision-defective Saccharomyces cerevisiae mutants does not rely on molecular recombination.
    • DNA gap filling during postreplication repair proceeds independently of extensive pyrimidine dimer transfer.
    • These findings suggest alternative mechanisms for bypassing or repairing UV-induced DNA damage in the absence of excision repair.