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Replicative intermediates in UV-irradiated simian virus 40.

J M Clark, P C Hanawalt

    Mutation Research
    |July 1, 1984
    PubMed
    Summary
    This summary is machine-generated.

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    Simian virus 40 (SV40) replication in UV-damaged DNA reveals that replication forks pause at pyrimidine dimers. Daughter strand continuity is eventually restored, suggesting DNA repair mechanisms are involved in processing UV-damaged DNA.

    Area of Science:

    • Molecular Biology
    • Virology
    • DNA Repair

    Background:

    • DNA damage, particularly from UV radiation, poses a significant threat to cellular integrity.
    • Understanding how DNA replication proceeds in the presence of DNA lesions is crucial for cell survival and preventing mutations.

    Purpose of the Study:

    • To investigate the mechanisms of DNA replication fork progression across UV-induced DNA damage.
    • To elucidate the role of Simian virus 40 (SV40) as a model system for studying DNA repair during replication in mammalian cells.

    Main Methods:

    • Utilized a synchronized SV40 DNA replication system in monkey kidney cells.
    • Employed UV irradiation to introduce pyrimidine dimers into viral DNA.
    • Isolated and analyzed replicative intermediates (RI) using density gradients and chromatography.

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  • Assessed DNA strand lengths and dimer presence in newly synthesized DNA.

    • Main Results:

    • Replication forks were temporarily blocked by pyrimidine dimers on the leading strand.
    • Daughter strand continuity opposite dimers was eventually established, despite no significant dimer excision.
    • Newly synthesized DNA strands initially showed a size bias, which resolved over time.
    • Form I DNA molecules containing dimers in daughter strands were generated late after irradiation.

    Conclusions:

    • Replication fork stalling at UV-induced dimers is a transient event.
    • DNA repair mechanisms, potentially involving trans-dimer synthesis and strand exchange, contribute to the completion of replication of damaged DNA.
    • SV40 serves as a valuable model for studying complex DNA replication and repair processes in mammalian systems.