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A Fluorescence-based Method to Study Bacterial Gene Regulation in Infected Tissues
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AraC-Type Regulator Rbf Controls the Staphylococcus epidermidis Biofilm Phenotype by Negatively Regulating the

Sarah E Rowe1, Christopher Campbell2, Colm Lowry2

  • 1Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland School of Biomolecular and Biomedical Science and Conway Institute, University College Dublin, Belfield, Dublin, Ireland.

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|August 10, 2016
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The transcriptional regulator Rbf controls Staphylococcus epidermidis biofilm formation by downregulating SarR and indirectly repressing SarX. This intricate regulation fine-tunes polysaccharide intercellular adhesin (PIA) production, crucial for medical device infections.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Medical Mycology

Background:

  • Staphylococcus epidermidis is a major cause of medical device-related infections due to its biofilm-forming capacity.
  • Polysaccharide intercellular adhesin (PIA) production, encoded by the icaADBC operon, is a key mechanism for S. epidermidis biofilm formation.
  • The regulation of icaADBC expression is complex and critical for controlling biofilm phenotypes.

Purpose of the Study:

  • To elucidate the role of the transcriptional regulator Rbf in the regulation of icaADBC expression and PIA production in Staphylococcus epidermidis.
  • To identify the specific mechanisms by which Rbf influences icaADBC transcription and biofilm formation.
  • To understand the interplay between Rbf and other known regulators of the ica operon.

Main Methods:

  • Electrophoretic mobility shift assays (EMSAs) to assess Rbf binding to promoter regions.
  • Reverse transcription (RT-PCR) to quantify gene expression levels.
  • Construction and analysis of rbf sarR double mutants to evaluate genetic interactions.

Main Results:

  • Rbf activates icaADBC transcription and PIA production indirectly, as it does not bind the ica operon promoter.
  • Rbf directly represses the transcription of sarR, a known repressor of the ica operon.
  • Rbf also indirectly represses the transcription of SarX, an activator of the ica operon, with SarR downregulation having a dominant effect.

Conclusions:

  • Rbf plays a significant role in regulating S. epidermidis biofilm formation by modulating the activity of SarR and SarX.
  • The Rbf-SarR-SarX regulatory axis provides a fine-tuning mechanism for icaADBC expression and biofilm phenotype.
  • Understanding this regulatory network is crucial for developing strategies to combat S. epidermidis biofilm-associated infections.