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Replication control and switch-off function as observed with a mini-F factor plasmid

H Tsutsui, K Matsubara

    Journal of Bacteriology
    |August 1, 1981
    PubMed
    Summary
    This summary is machine-generated.

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    This study reveals a switch-off mechanism controlling mini-F plasmid replication initiation. High copy numbers of mini-F trigger this mechanism, reducing deoxyribonucleic acid synthesis, supporting the inhibitor dilution model.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Plasmid Biology

    Background:

    • Mini-F is a 9,000 base pair fragment of the F plasmid, essential for replication maintenance and control.
    • Mini-F has a low copy number (1-2 per chromosome), while ColE1 has a high copy number (16-20 per chromosome).
    • Composite plasmids can exhibit replication characteristics of either parent plasmid.

    Purpose of the Study:

    • To investigate the control mechanisms governing plasmid replication.
    • To examine the transition between high and low copy number replication in composite plasmids.
    • To elucidate the switch-off mechanism for mini-F replication initiation.

    Main Methods:

    • Constructed a composite plasmid by joining mini-F and ColE1.
    • Inhibited ColE1 replication using deoxyribonucleic acid polymerase I inactivation.

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  • Observed replication characteristics and copy numbers under different conditions, including using ppF (mini-F replicon in phage lambda head).

    • Main Results:

    • Under normal conditions, the composite plasmid replicated with ColE1's high copy number.
    • Upon ColE1 inhibition, replication shifted to mini-F's low copy number characteristics.
    • Mini-F replication initiation was blocked at high copy numbers and restored upon reduction, indicating a copy number-dependent switch-off mechanism.

    Conclusions:

    • A copy number-dependent switch-off mechanism actively regulates mini-F replication initiation.
    • This mechanism is consistent with the 'inhibitor dilution model' for plasmid replication control.
    • The study provides insights into the nature of the hypothetical inhibitor involved in this control system.