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Incompatibility and IncFII plasmid replication control.

R H Rownd, D D Womble, X N Dong

    Basic Life Sciences
    |January 1, 1985
    PubMed
    Summary
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    Plasmid NR1 replication is controlled by a small RNA molecule that binds to larger RNA transcripts, inhibiting protein production and regulating plasmid copy number. This RNA-RNA interaction governs plasmid replication initiation.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Biochemistry

    Background:

    • Plasmid NR1 DNA encodes replication control and incompatibility functions.
    • Transcription occurs bidirectionally from the plasmid DNA template.

    Purpose of the Study:

    • To elucidate the mechanism of plasmid NR1 replication control and incompatibility.
    • To characterize the roles of different RNA transcripts in plasmid regulation.

    Main Methods:

    • In vivo and in vitro RNA transcription analysis.
    • Quantitative hybridization experiments to estimate intracellular RNA concentrations.
    • Mass action law predictions based on RNA-RNA duplex formation and base-stacking free energies.

    Main Results:

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  • Bidirectional transcription yields large messenger RNAs (rightward) and a small regulatory RNA (leftward).
  • The small leftward transcript acts as an incompatibility product, inversely regulating rightward transcript translation and replication initiation.
  • RNA-RNA duplex formation between the small and large transcripts inhibits replication protein synthesis.

    • Conclusions:

    • The small incompatibility RNA controls plasmid NR1 replication by inhibiting translation of essential replication proteins.
    • The mechanism involves RNA-RNA duplex formation, predictable by mass action laws and thermodynamic principles.
    • Experimental RNA concentrations align with theoretical predictions for duplex formation and regulatory function.