The eukaryotic-type serine/threonine protein kinase Stk is required for biofilm formation and virulence in Staphylococcus epidermidis

PLoS One. 2011;6(9):e25380. doi: 10.1371/journal.pone.0025380. Epub 2011 Sep 23.

Abstract

Background: Serine/threonine kinases are involved in gene regulation and signal transduction in prokaryotes and eukaryotes. Here, we investigated the role of the serine/threonine kinase Stk in the opportunistic pathogen Staphylococcus epidermidis.

Methodology/principal findings: We constructed an isogenic stk mutant of a biofilm-forming clinical S. epidermidis isolate. Presence of stk was important for biofilm formation in vitro and virulence in a murine subcutaneous foreign body infection model. Furthermore, the stk mutant exhibited phenotypes indicating an impact of stk on metabolic pathways. Using different constructs for the genetic complementation of the stk mutant strain with full-length Stk or specific Stk domains, we determined that the Stk intracellular kinase domain is important for biofilm formation and regulation of purine metabolism. Site-specific inactivation of the Stk kinase domain led to defective biofilm formation, in further support of the notion that the kinase activity of Stk regulates biofilm formation of S. epidermidis. According to immunological detection of the biofilm exopolysaccharide PIA and real-time PCR of the PIA biosynthesis genes, the impact of stk on biofilm formation is mediated, at least in part, by a strong influence on PIA expression.

Conclusions: Our study identifies Stk as an important regulator of biofilm formation and virulence of S. epidermidis, with additional involvement in purine metabolism and the bacterial stress response.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Biofilms / growth & development*
  • Gene Expression Regulation, Bacterial / genetics
  • Gene Expression Regulation, Bacterial / physiology
  • Immunoblotting
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mutagenesis, Site-Directed
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Staphylococcal Infections / enzymology
  • Staphylococcal Infections / genetics
  • Staphylococcus epidermidis / enzymology*
  • Staphylococcus epidermidis / genetics
  • Staphylococcus epidermidis / growth & development*
  • Staphylococcus epidermidis / pathogenicity*
  • Virulence / genetics
  • Virulence / physiology*

Substances

  • Bacterial Proteins
  • Protein Serine-Threonine Kinases