title: N-acylhomoserine lactones antagonize virulence gene expression and quorum sensing in Staphylococcus aureus.
abstract: Many gram-negative bacteria employ N-acylhomoserine lactone (AHL)-mediated quorum sensing to control virulence. To determine whether gram-positive bacteria such as Staphylococcus aureus respond to AHLs, we used a growth-dependent lux reporter fusion. Exposure of S. aureus to different AHLs revealed that 3-oxo-substituted AHLs with C10 to C14 acyl chains inhibited light output and growth in a concentration-dependent manner, while short-chain AHLs had no effect. N-(3-Oxododecanoyl)-L-homoserine lactone (3-oxo-C12-HSL) inhibited the production of exotoxins and cell wall fibronectin-binding proteins but enhanced protein A expression. Since these processes are reciprocally regulated via the S. aureus agr quorum-sensing system, which in turn, is regulated via sar, we examined the effect of AHLs on sarA and agr. At sub-growth-inhibitory concentrations of 3-oxo-C12-HSL, both sarA expression and agr expression were inhibited, indicating that the action of 3-oxo-C12-HSL is mediated at least in part through antagonism of quorum sensing in S. aureus. Spent culture supernatants from Pseudomonas aeruginosa, which produces both 3-oxo-C12-HSL and N-butanoyl-homoserine lactone (C4-HSL), also inhibited agr expression, although C4-HSL itself was inactive in this assay. Since quorum sensing in S. aureus depends on the activities of membrane-associated proteins, such as AgrB, AgrC, and AgrD, we investigated whether AHLs perturbed S. aureus membrane functionality by determining their influence on the membrane dipole potential. From the binding curves obtained, a dissociation constant of 7 muM was obtained for 3-oxo-C12-HSL, indicating the presence of a specific saturable receptor, whereas no binding was observed for C4-HSL. These data demonstrate that long-chain 3-oxo-substituted AHLs, such as 3-oxo-C12-HSL, are capable of interacting with the S. aureus cytoplasmic membrane in a saturable, specific manner and at sub-growth-inhibitory concentrations, down-regulating exotoxin production and both sarA and agr expression.
authors.affiliation: Institute of Infection, Immunity and Inflammation, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, United Kingdom.
authors.forename: Saara
authors.initials: S
authors.lastname: Qazi
authors.forename: Barry
authors.initials: B
authors.lastname: Middleton
authors.forename: Siti Hanna
authors.initials: SH
authors.lastname: Muharram
authors.forename: Alan
authors.initials: A
authors.lastname: Cockayne
authors.forename: Philip
authors.initials: P
authors.lastname: Hill
authors.forename: Paul
authors.initials: P
authors.lastname: O'Shea
authors.forename: Siri Ram
authors.initials: SR
authors.lastname: Chhabra
authors.forename: Miguel
authors.initials: M
authors.lastname: Cámara
authors.forename: Paul
authors.initials: P
authors.lastname: Williams
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D015536:Down-Regulation; D005098:Exotoxins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006714:Homoserine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D011550:Pseudomonas aeruginosa / Q000254:growth & development / Q000378:metabolism; D015398:Signal Transduction; D013211:Staphylococcus aureus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism / Q000472:pathogenicity*; D015534:Trans-Activators / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014774:Virulence
chemical_list: C095698:Agr protein, Staphylococcus aureus; D001426:Bacterial Proteins; D005098:Exotoxins; C109860:N-(3-oxododecanoyl)homoserine lactone; C092312:N-butyrylhomoserine lactone; C492107:SarA protein, Staphylococcus aureus; D015534:Trans-Activators; C088386:homoserine lactone; D006714:Homoserine; D015107:4-Butyrolactone