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DNA-Binding Proteins Regulating pIP501 Transfer and Replication.

Elisabeth Grohmann1, Nikolaus Goessweiner-Mohr2, Sabine Brantl3

  • 1Division of Infectious Diseases, University Medical Center FreiburgFreiburg im Breisgau, Germany; Life Sciences and Technology, Beuth University of Applied Sciences BerlinBerlin, Germany.

Frontiers in Molecular Biosciences
|August 27, 2016
PubMed
Summary
This summary is machine-generated.

The pIP501 plasmid, a key tool in studying bacterial genetics, replicates via a theta mechanism and transfers using a broad-host-range type IV secretion system. Its replication and transfer are tightly regulated by specific proteins and RNA molecules.

Keywords:
broad-host-rangeconjugative plasmidcopy number controlreplicationtransfer controltype IV secretion

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Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • pIP501 is a Gram-positive model plasmid utilized for studying plasmid replication and conjugative transfer.
  • It is frequently found in clinical strains of Enterococcus faecalis and Enterococcus faecium, conferring multiple antibiotic resistances.

Purpose of the Study:

  • To elucidate the regulatory mechanisms governing the replication and conjugative transfer of the pIP501 plasmid.
  • To understand the roles of key proteins and RNA molecules in controlling plasmid copy number and transfer efficiency.

Main Methods:

  • Analysis of plasmid replication using theta mechanism.
  • Investigation of regulatory elements including repR, oriR, CopR, and RNAIII.
  • Characterization of the type IV secretion system (T4SS) operon and its regulatory protein TraN.

Main Results:

  • Plasmid replication is initiated by RepR protein at oriR and regulated by CopR and antisense RNAIII, with deletions increasing copy number.
  • The conjugative transfer of pIP501 relies on a broad-host-range T4SS, transferable to various Gram-positive and even Gram-negative bacteria.
  • TraA protein regulates T4SS expression, while TraN acts as a repressor of pIP501 transfer, with its absence significantly increasing transfer rates.

Conclusions:

  • The pIP501 plasmid possesses intricate regulatory systems for both replication and conjugative transfer.
  • Understanding these mechanisms is crucial for studying antibiotic resistance dissemination and developing novel genetic tools.