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Two replication initiation sites on R-plasmid DNA.

J H Crosa, L K Luttropp, F Heffron

    Molecular & General Genetics : MGG
    |September 15, 1975
    PubMed
    Summary
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    Replication of R 6 K plasmid DNA initiates at two distinct sites. Some molecules utilize both initiation sites simultaneously during replication.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • The conjugative R-plasmid R 6 K is a key element in bacterial genetic exchange.
    • Understanding plasmid replication mechanisms is crucial for molecular biology and genetics.

    Purpose of the Study:

    • To investigate the replication initiation sites of a deletion mutant of the R 6 K plasmid.
    • To characterize the spatial and functional relationship of these initiation sites.

    Main Methods:

    • Cleavage of R 6 K plasmid DNA with EcoRI restriction endonuclease.
    • Electron microscopy and molecular measurements of EcoRI-treated replicative intermediate molecules.

    Main Results:

    • Replicating R 6 K plasmid DNA molecules were cleaved at a single site by EcoRI.

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  • Replication initiation was found to occur at two specific sites on the plasmid DNA.
  • These sites are located at approximately 23% and 39% of the total length from the EcoRI cleavage site.
  • Approximately 5% of replicating molecules initiated replication from both sites concurrently.

    • Conclusions:

    • The R 6 K plasmid possesses multiple, distinct replication origins.
    • The simultaneous use of both initiation sites by a subset of molecules suggests complex regulatory mechanisms governing plasmid replication.