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Replication control for pT181, an indirectly regulated plasmid.

R P Novick, S J Projan, C C Kumar

    Basic Life Sciences
    |January 1, 1985
    PubMed
    Summary
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    The Staphylococcus aureus plasmid pT181 uses a negative regulatory system involving two countertranscripts to control replication protein synthesis, differing from direct regulation seen in E. coli plasmids.

    Area of Science:

    • Molecular Biology
    • Microbiology
    • Genetics

    Background:

    • The Staphylococcus aureus plasmid pT181 is a 4,437 bp tetracycline resistance plasmid with a copy number of approximately 22 per cell.
    • Plasmid replication control mechanisms vary, with direct and indirect regulation observed in different plasmid groups.

    Purpose of the Study:

    • To elucidate the replication control mechanism of the Staphylococcus aureus plasmid pT181.
    • To compare the replication regulation of pT181 with that of Co1E1 and IncFII plasmids in Escherichia coli.

    Main Methods:

    • Sequence analysis of the pT181 replicon.
    • Identification and characterization of regulatory transcripts (RNA I and RNA II).
    • Analysis of replication control mechanisms through mutant studies.

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    Main Results:

    • pT181 replication is regulated by the 35-kd RepC protein, essential for replication, with its origin located within the repC coding sequence.
    • A negative regulatory system involving two countertranscripts (RNA I and RNA II) controls RepC synthesis, likely by interfering with repC mRNA translation.
    • pT181 exhibits indirect replication regulation, where the RepC protein is trans-active, contrasting with direct regulation in Co1E1 and IncFII plasmids.

    Conclusions:

    • pT181 employs an indirect replication control mechanism involving countertranscripts and the RepC protein.
    • Differences in regulation (direct vs. indirect) lead to distinct behaviors regarding mutant coexistence and incompatibility group formation.