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Infection and Immunity, July 2004, p. 3838-3848, Vol. 72, No. 7
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.7.3838-3848.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

RsbU-Dependent Regulation of Staphylococcus epidermidis Biofilm Formation Is Mediated via the Alternative Sigma Factor {sigma}B by Repression of the Negative Regulator Gene icaR

Johannes K.-M. Knobloch,* Sebastian Jäger, Matthias A. Horstkotte, Holger Rohde, and Dietrich Mack

Universitätsklinikum Hamburg-Eppendorf, Zentrum für Klinisch-Theoretische Medizin I, Institut für Infektionsmedizin, D-20246 Hamburg, Germany

Received 14 October 2003/ Returned for modification 27 January 2004/ Accepted 2 April 2004

Transposon mutagenesis of rsbU leads to a biofilm-negative phenotype in Staphylococcus epidermidis. However, the pathway of this regulatory mechanism was unknown. To investigate the role of RsbU in the regulation of the alternative sigma factor {sigma}B and biofilm formation, we generated different mutants of the {sigma}B operon in S. epidermidis strains 1457 and 8400. The genes rsbU, rsbV, rsbW, and sigB, as well as the regulatory cascade rsbUVW and the entire {sigma}B operon, were deleted. Transcriptional analysis of sarA and the {sigma}B-dependent gene asp23 revealed the functions of RsbU and RsbV as positive regulators and of RsbW as a negative regulator of {sigma}B activity, indicating regulation of {sigma}B activity similar to that characterized for Bacillus subtilis and Staphylococcus aureus. Phenotypic characterization of the mutants revealed that the dramatic decrease of biofilm formation in rsbU mutants is mediated via {sigma}B, indicating a crucial role for {sigma}B in S. epidermidis pathogenesis. However, biofilm formation in mutants defective in {sigma}B or its function could be restored in the presence of subinhibitory ethanol concentrations. Transcriptional analysis revealed that icaR is up-regulated in mutants lacking {sigma}B function but that icaA transcription is down-regulated in these mutants, indicating a {sigma}B-dependent regulatory intermediate negatively regulating IcaR. Supplementation of growth media with ethanol decreased icaR transcription, leading to increased icaA transcription and a biofilm-positive phenotype, indicating that the ethanol-dependent induction of biofilm formation is mediated by IcaR. This icaR-dependent regulation under ethanol induction is mediated in a {sigma}B-independent manner, suggesting at least one additional regulatory intermediate in the biofilm formation of S. epidermidis.

* Corresponding author. Mailing address: Universitätsklinikum Hamburg-Eppendorf, Zentrum für Klinisch-Theoretische Medizin I, Institut für Infektionsmedizin, Martinistr. 52, D-20246 Hamburg, Germany. Phone: 49 40 42803 3147. Fax: 49 40 42803 4881. E-mail: knobloch{at}uke.uni-hamburg.de.

Editor: V. J. DiRita

Infection and Immunity, July 2004, p. 3838-3848, Vol. 72, No. 7
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.7.3838-3848.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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