title: A unique regulator contributes to quorum sensing and virulence in Burkholderia cenocepacia.
abstract: Burkholderia cenocepacia causes chronic and life-threatening respiratory infections in immunocompromized people. The B. cenocepacia N-acyl-homoserine lactone (AHL)-dependent quorum sensing system relies on the production of AHLs by the synthases CepI and CciI while CepR, CciR and CepR2 control expression of many genes important for pathogenesis. Downstream from, and co-transcribed with cepI, lies BCAM1871 encoding a hypothetical protein that was uncharacterized prior to this study. Orthologs of B. cenocepacia BCAM1871 are uniquely found in Burkholderia spp and are conserved in their genomic locations in pathogenic Burkholderia. We observed significant effects on AHL activity upon mutation or overexpression of BCAM1871, although these effects were more subtle than those observed for CepI indicating BCAM1871 acts as an enhancer of AHL activity. Transcription of cepI, cepR and cciIR was significantly reduced in the BCAM1871 mutant. Swimming and swarming motilities as well as transcription of fliC, encoding flagellin, were significantly reduced in the BCAM1871 mutant. Protease activity and transcription of zmpA and zmpB, encoding extracellular zinc metalloproteases, were undetectable in the BCAM1871 mutant indicating a more significant effect of mutating BCAM1871 than cepI. Exogenous addition of OHL restored cepI, cepR and fliC transcription but had no effect on motility, protease activity or zmpA or zmpB transcription suggesting AHL-independent effects. The BCAM1871 mutant exhibited significantly reduced virulence in rat chronic respiratory and nematode infection models. Gene expression and phenotypic assays as well as vertebrate and invertebrate infection models showed that BCAM1871 significantly contributes to pathogenesis in B. cenocepacia.
authors.affiliation: Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Eoin P
authors.initials: EP
authors.lastname: O'Grady
authors.forename: Duber F
authors.initials: DF
authors.lastname: Viteri
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Sokol
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D057508:Burkholderia cenocepacia / Q000472:pathogenicity* / Q000502:physiology; D017173:Caenorhabditis elegans / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics / Q000502:physiology*; D056426:Genetic Loci / Q000235:genetics*; D053208:Kaplan-Meier Estimate; D008025:Ligases / Q000235:genetics / Q000378:metabolism; D008666:Metalloendopeptidases / Q000378:metabolism; D016366:Open Reading Frames / Q000235:genetics*; D053038:Quorum Sensing / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D012141:Respiratory Tract Infections / Q000378:metabolism* / Q000382:microbiology; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D001426:Bacterial Proteins; C117246:CepR protein, Burkholderia; D017931:DNA Primers; C088386:homoserine lactone; D008666:Metalloendopeptidases; C502465:ZmpA protein, Burkholderia cenocepacia; D008025:Ligases; C449816:N-acylhomoserine lactone synthase; D015107:4-Butyrolactone