title: SigmaB and SarA independently regulate polysaccharide intercellular adhesin production in Staphylococcus epidermidis.
abstract: The production of polysaccharide intercellular adhesin (PIA) is an essential process in foreign body infections mediated by Staphylococcus epidermidis. Transcriptional regulation of the icaADBC operon, the genes responsible for production of enzymes that synthesize PIA, is multi-factorial and involves at least SarA and sigmaB. Transcriptional and promoter fusion studies revealed that the decreased transcription of the icaADBC operon observed in a S. epidermidis 1457 sigB mutant is not mediated through a direct interaction of sigmaB-RNA polymerase at the icaADBC promoter region but instead through the upregulation of IcaR, a known repressor of icaADBC transcription. Transcriptional analysis of a 1457 sigB-icaR double mutant confirmed that the decreased icaADBC transcript in 1457 sigB is IcaR dependent. Furthermore, primer extension studies suggest that the icaR promoter appears to be sigmaA dependent, suggesting that sigmaB indirectly controls icaR transcription through an unknown pathway. In addition, it was confirmed that the loss of SarA results in the loss of icaADBC transcription and PIA production in S. epidermidis. It was further demonstrated, through the over-production of SarA in 1457 sigB, that the loss of sarP1 promoter activity in 1457 sigB has little or no effect on the loss of PIA production in this mutant. Finally, it was demonstrated that PIA production could be restored in both 1457 sigB and 1457 sarA by complementing these mutants with a full-length icaADBC operon controlled by a cadmium-inducible noncognate promoter. It is concluded that sigmaB and SarA operate independently of each other to regulate PIA production and biofilm development in S. epidermidis.
authors.affiliation: Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68198-6280, USA.
authors.forename: L D
authors.initials: LD
authors.lastname: Handke
authors.forename: S R
authors.initials: SR
authors.lastname: Slater
authors.forename: K M
authors.initials: KM
authors.lastname: Conlon
authors.forename: Sinead T
authors.initials: ST
authors.lastname: O'Donnell
authors.forename: M E
authors.initials: ME
authors.lastname: Olson
authors.forename: K A
authors.initials: KA
authors.lastname: Bryant
authors.forename: M E
authors.initials: ME
authors.lastname: Rupp
authors.forename: J P
authors.initials: JP
authors.lastname: O'Gara
authors.forename: P D
authors.initials: PD
authors.lastname: Fey
mesh_terms: D001426:Bacterial Proteins / Q000502:physiology*; D018441:Biofilms / Q000254:growth & development*; D009876:Operon / Q000502:physiology; D011135:Polysaccharides, Bacterial / Q000378:metabolism*; D012808:Sigma Factor / Q000502:physiology*; D013212:Staphylococcus epidermidis / Q000472:pathogenicity / Q000502:physiology*; D015534:Trans-Activators / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D011135:Polysaccharides, Bacterial; C088021:SarA protein, bacterial; C079011:SigB protein, Bacteria; D012808:Sigma Factor; D015534:Trans-Activators; C113579:polysaccharide intercellular adhesin