title: Essential role for the major autolysin in the fibronectin-binding protein-mediated Staphylococcus aureus biofilm phenotype.
abstract: Staphylococcus aureus clinical isolates are capable of producing at least two distinct types of biofilm mediated by the fibronectin-binding proteins (FnBPs) or the icaADBC-encoded polysaccharide intercellular adhesin (PIA). Deletion of the major autolysin gene atl reduced primary attachment rates and impaired FnBP-dependent biofilm production on hydrophilic polystyrene in 12 clinical methicillin-resistant S. aureus (MRSA) isolates but had no effect on PIA-dependent biofilm production by 9 methicillin-susceptible S. aureus (MSSA) isolates. In contrast, Atl was required for both FnBP- and PIA-mediated biofilm development on hydrophobic polystyrene. Here we investigated the role of Atl in biofilm production on hydrophilic polystyrene. The alternative sigma factor σ(B), which represses RNAIII expression and extracellular protease production, was required for FnBP- but not PIA-dependent biofilm development. Furthermore, mutation of the agr locus enhanced FnBP-dependent biofilm development, whereas a sarA mutation, which increases protease production, blocked FnBP-mediated biofilm development. Mutation of sigB in MRSA isolate BH1CC lowered primary attachment rates, in part via reduced atl transcription. Posttranslational activation or inhibition of Atl activity with phenylmethylsulfonyl fluoride and polyanethole sodium sulfonate or mutation of the Atl amidase active site interfered with lytic activity and biofilm development. Consistent with these observations, extracellular DNA was important for the early stages of Atl/FnBP-dependent biofilm development. Further analysis of atl regulation revealed that atlR encodes a transcriptional repressor of the major autolysin and that an atlR::Tc(r) mutation in BH1CC enhanced biofilm-forming capacity. These data reveal an essential role for the major autolysin in the early events of the FnBP-dependent S. aureus biofilm phenotype.
authors.affiliation: School of Biomolecular and Biomedical Science and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Patrick
authors.initials: P
authors.lastname: Houston
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Rowe
authors.forename: Clarissa
authors.initials: C
authors.lastname: Pozzi
authors.forename: Elaine M
authors.initials: EM
authors.lastname: Waters
authors.forename: James P
authors.initials: JP
authors.lastname: O'Gara
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D018441:Biofilms; D024202:Electrophoretic Mobility Shift Assay; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000502:physiology*; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000378:metabolism*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012808:Sigma Factor / Q000378:metabolism; D013211:Staphylococcus aureus / Q000502:physiology*; D015534:Trans-Activators / Q000378:metabolism
chemical_list: D018829:Adhesins, Bacterial; C095698:Agr protein, Staphylococcus aureus; D001426:Bacterial Proteins; C492107:SarA protein, Staphylococcus aureus; D012808:Sigma Factor; D015534:Trans-Activators; C078635:fibronectin-binding proteins, bacterial; D009238:N-Acetylmuramoyl-L-alanine Amidase