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Coding sequence for the pT181 repC product: a plasmid-coded protein uniquely required for replication.

R P Novick, G K Adler, S Majumder

    Proceedings of the National Academy of Sciences of the United States of America
    |July 1, 1982
    PubMed
    Summary
    This summary is machine-generated.

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    The Staphylococcus aureus plasmid pT181 requires a specific protein, RepC, for replication. This study confirms RepC

    Area of Science:

    • Molecular Biology
    • Microbiology
    • Genetics

    Background:

    • The pT181 plasmid from Staphylococcus aureus confers tetracycline resistance.
    • Previous studies proposed a diffusible plasmid product essential for replication based on thermosensitive mutants (Tsr).

    Purpose of the Study:

    • To investigate the nature of the proposed diffusible product required for pT181 plasmid replication.
    • To identify and characterize the gene responsible for pT181 plasmid maintenance.

    Main Methods:

    • Genetic complementation analysis of thermosensitive mutants.
    • Biochemical confirmation of replication defects.
    • DNA sequence analysis to identify open reading frames.
    • In vitro replication assays using cell-free extracts.

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    Main Results:

    • Thermosensitive mutants (Tsr) exhibit primary replication defects, confirming the proposed model.
    • All five mutations map to a single cistron, designated repC.
    • DNA sequencing revealed an open reading frame encoding a 313-amino acid protein (RepC).
    • Cell-free extracts demonstrated the requirement of a pT181-coded protein of ~313 amino acids for replication, identified as the RepC product.

    Conclusions:

    • The RepC protein is essential for pT181 plasmid replication in Staphylococcus aureus.
    • Replication rate of pT181 may be regulated by the synthesis of the RepC protein.