Biofilm Formation by Staphylococcus epidermidis Depends on Functional RsbU, an Activator of the sigB Operon: Differential Activation Mechanisms Due to Ethanol and Salt Stress

Abstract

Staphylococcus epidermidis is a common pathogen in medical device-associated infections. Its major pathogenetic factor is the ability to form adherent biofilms. The polysaccharide intercellular adhesin (PIA), which is synthesized by the products of the icaADBC gene cluster, is essential for biofilm accumulation. In the present study, we characterized the gene locus inactivated by Tn 917 insertions of two isogenic, icaADBC -independent, biofilm-negative mutants, M15 and M19, of the biofilm-producing bacterium S. epidermidis 1457. The insertion site was the same in both of the mutants and was located in the first gene, rsbU , of an operon highly homologous to the sigB operons of Staphylococcus aureus and Bacillus subtilis . Supplementation of Trypticase soy broth with NaCl (TSB NaCl ) or ethanol (TSB EtOH ), both of which are known activators of sigB , led to increased biofilm formation and PIA synthesis by S. epidermidis 1457. Insertion of Tn 917 into rsbU , a positive regulator of alternative sigma factor ς B , led to a biofilm-negative phenotype and almost undetectable PIA production. Interestingly, in TSB EtOH , the mutants were enabled to form a biofilm again with phenotypes similar to those of the wild type. In TSB NaCl , the mutants still displayed a biofilm-negative phenotype. No difference in primary attachment between the mutants and the wild type was observed. Similar phenotypic changes were observed after transfer of the Tn 917 insertion of mutant M15 to the independent and biofilm-producing strain S. epidermidis 8400. In 11 clinical S. epidermidis strains, a restriction fragment length polymorphism of the sigB operon was detected which was independent of the presence of the icaADBC locus and a biofilm-positive phenotype. Obviously, different mechanisms are operative in the regulation of PIA expression in stationary phase and under stress induced by salt or ethanol.