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Purified bacteriophage lambda O protein binds to four repeating sequences at the lambda replication origin.

T Tsurimoto, K Matsubara

    Nucleic Acids Research
    |April 24, 1981
    PubMed
    Summary
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    Bacteriophage lambda O protein initiates DNA replication by binding to the origin (ori) sequence. This protein binds to a 95 base pair region containing four repeats, likely in dimeric form, to activate DNA synthesis.

    Area of Science:

    • Molecular Biology
    • Virology
    • Genetics

    Background:

    • Bacteriophage lambda DNA replication initiation is crucial for viral propagation.
    • The bacteriophage lambda O protein is a key initiator factor.
    • Interaction between O protein and the origin (ori) sequence is essential for replication initiation.

    Purpose of the Study:

    • To investigate the binding interaction between bacteriophage lambda O protein and its specific DNA origin (ori).
    • To characterize the binding site and stoichiometry of O protein on the lambda ori DNA sequence.
    • To elucidate the mechanism of O protein-mediated ori activation for DNA replication.

    Main Methods:

    • Purification of bacteriophage lambda O protein to near homogeneity.
    • Nuclease protection assays to map O protein binding sites on ori DNA.

    Related Experiment Videos

  • Analysis of O protein binding as a function of protein concentration.

    • Main Results:

    • The O protein binds to a 95 base pair region within the lambda ori.
    • This 95 bp region contains four tandemly arranged 19 bp repeating sequences.
    • O protein binding is concentration-dependent, with inner repeats binding at lower concentrations and outer repeats at higher concentrations.
    • Evidence suggests O protein binds in dimeric form, with four dimers potentially occupying the 95 bp region.

    Conclusions:

    • The bacteriophage lambda O protein specifically binds to a defined region of the ori sequence.
    • The binding involves multiple O protein dimers interacting with repeating units within the ori.
    • This precise interaction is critical for the activation of lambda DNA replication initiation.