N-Octanoylhomoserine lactone signalling mediated by the BpsI-BpsR quorum sensing system plays a major role in biofilm formation of Burkholderia pseudomallei.
The genome of Burkholderia pseudomallei encodes three acylhomoserine lactone (AHL) quorum sensing systems, each comprising an AHL synthase and a signal receptor/regulator. The BpsI-BpsR system produces N-octanoylhomoserine lactone (C8HL) and is positively auto-regulated by its AHL product. The products of the remaining two systems have not been identified. In this study, tandem MS was used to identify and quantify the AHL species produced by three clinical B. pseudomallei isolates - KHW, K96243 and H11 - three isogenic KHW mutants that each contain a null mutation in an AHL synthase gene, and recombinant Escherichia coli heterologously expressing each of the three B. pseudomallei AHL synthase genes. BpsI synthesized predominantly C8HL, which accounted for more than 95 % of the extracellular AHLs produced in stationary-phase KHW cultures. The major products of BpsI(2) and BpsI(3) were N-(3-hydroxy-octanoyl)homoserine lactone (OHC8HL) and N-(3-hydroxy-decanoyl)homoserine lactone, respectively, and their corresponding transcriptional regulators, BpsR(2) and BpsR(3), were capable of driving reporter gene expression in the presence of these cognate lactones. Formation of biofilm by B. pseudomallei KHW was severely impaired in mutants lacking either BpsI or BpsR but could be restored to near wild-type levels by exogenous C8HL. BpsI(2) was not required, and BpsI(3) was partially required for biofilm formation. Unlike the bpsI mutant, biofilm formation in the bpsI(3) mutant could not be restored to wild-type levels in the presence of OHC8HL, the product of BpsI(3). C8HL and OHC8HL had opposite effects on biofilm formation; exogenous C8HL enhanced biofilm formation in both the bpsI(3) mutant and wild-type KHW while exogenous OHC8HL suppressed the formation of biofilm in the same strains. We propose that exogenous OHC8HL antagonizes biofilm formation in B. pseudomallei, possibly by competing with endogenous C8HL for binding to BpsR.
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 8 Medical Drive, 117597 Singapore.
Akshamal Mihiranga
AM
Gamage
Life Science Institute, National University of Singapore, 28 Medical Drive, 117456 Singapore.
Guanghou
G
Shui
Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, 117543 Singapore.|Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 8 Medical Drive, 117597 Singapore.
Markus R
MR
Wenk
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 8 Medical Drive, 117597 Singapore.
Kim Lee
KL
Chua
D018441:Biofilms / Q000254:growth & development*; D016957:Burkholderia pseudomallei / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D006714:Homoserine / Q000031:analogs & derivatives* / Q000378:metabolism; D007783:Lactones / Q000378:metabolism*; D008025:Ligases / Q000235:genetics / Q000378:metabolism*; D053038:Quorum Sensing; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D007783:Lactones; C091088:N-octanoylhomoserine lactone; D011994:Recombinant Proteins; D014157:Transcription Factors; D006714:Homoserine; D008025:Ligases; C449816:N-acylhomoserine lactone synthase