title: The role of WlaRG, WlaTB and WlaTC in lipooligosaccharide synthesis by Campylobacter jejuni strain 81116.
abstract: Campylobacter jejuni is a major bacterial cause of gastroenteritis world-wide. C. jejuni produces a range of glycans including lipooligosaccharide (LOS), an important virulence factor. The genetic content of the LOS synthesis locus varies between C. jejuni strains and 19 classes have been described. Three LOS synthesis genes of C. jejuni strain 81116 (NCTC 11828), wlaRG, wlaTB and wlaTC were the focus of this study. WlaRG and the remaining two proteins of interest share sequence similarity to aminotransferases and glycosyltransferases, respectively. These genes were insertionally inactivated and phenotypically characterised. Each mutant produced truncated LOS. Mutants lacking WlaRG, WlaTB and WlaTC produced LOS with reduced immunogenicity. Both the wlaRG and wlaTC mutants were non-motile and aflagellate. In vitro invasion and adhesion assays revealed that the wlaRG, wlaTB and wlaTC mutants displayed reduced adherence to chicken embryo fibroblasts. All mutants were less invasive of human cells than 81116 confirming the role of intact LOS during invasion of human cells in vitro. Here we propose the general composition for the 81116 LOS core backbone based on capillary electrophoresis-mass spectrometry.
authors.affiliation: School of Applied Sciences, RMIT University, Melbourne, Bundoora, VIC 3083, Australia. karen.holden@csiro.au
authors.forename: Karen M
authors.initials: KM
authors.lastname: Holden
authors.forename: Michel
authors.initials: M
authors.lastname: Gilbert
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Coloe
authors.forename: Jianjun
authors.initials: J
authors.lastname: Li
authors.forename: Benjamin N
authors.initials: BN
authors.lastname: Fry
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016123:Campylobacter jejuni / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D002478:Cells, Cultured; D002642:Chick Embryo; D005347:Fibroblasts / Q000382:microbiology; D005407:Flagella / Q000502:physiology; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000276:immunology; D008124:Locomotion; D016254:Mutagenesis, Insertional; D017386:Sequence Homology, Amino Acid; D000637:Transaminases / Q000235:genetics / Q000378:metabolism*; D037521:Virulence Factors / Q000096:biosynthesis / Q000276:immunology
chemical_list: D001426:Bacterial Proteins; D008070:Lipopolysaccharides; D037521:Virulence Factors; C023023:lipid-linked oligosaccharides; D016695:Glycosyltransferases; D000637:Transaminases