title: Cj1136 is required for lipooligosaccharide biosynthesis, hyperinvasion, and chick colonization by Campylobacter jejuni.
abstract: Campylobacter jejuni is a major cause of bacterial food-borne enteritis worldwide, and invasion into intestinal epithelial cells is an important virulence mechanism. Recently we reported the identification of hyperinvasive C. jejuni strains and created a number of transposon mutants of one of these strains, some of which exhibited reduced invasion into INT-407 and Caco-2 cells. In one such mutant the transposon had inserted into a homologue of cj1136, which encodes a putative galactosyltransferase according to the annotation of the C. jejuni NCTC11168 genome. In the current study, we investigated the role of cj1136 in C. jejuni virulence, lipooligosaccharide (LOS) biosynthesis, and host colonization by targeted mutagenesis and complementation of the mutation. The cj1136 mutant showed a significant reduction in invasion into human intestinal epithelial cells compared to the wild-type strain 01/51. Invasion levels were partially restored on complementing the mutation. The inactivation of cj1136 resulted in the production of truncated LOS, while biosynthesis of a full-length LOS molecule was restored in the complemented strain. The cj1136 mutant showed an increase in sensitivity to the bile salts sodium taurocholate and sodium deoxycholate and significantly increased sensitivity to polymyxin B compared to the parental strain. Importantly, the ability of the mutant to colonize 1-day-old chicks was also significantly impaired. This study confirms that a putative galactosyltransferase encoded by cj1136 is involved in LOS biosynthesis and is important for C. jejuni virulence, as disruption of this gene and the resultant truncation of LOS affect both colonization in vivo and invasiveness in vitro.
authors.affiliation: School of Science and Technology, Nottingham Trent University, Nottingham, United Kingdom.
authors.forename: Muhammad Afzal
authors.initials: MA
authors.lastname: Javed
authors.forename: Shaun A
authors.initials: SA
authors.lastname: Cawthraw
authors.forename: Abiyad
authors.initials: A
authors.lastname: Baig
authors.forename: Jianjun
authors.initials: J
authors.lastname: Li
authors.forename: Alan
authors.initials: A
authors.lastname: McNally
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Oldfield
authors.forename: Diane G
authors.initials: DG
authors.lastname: Newell
authors.forename: Georgina
authors.initials: G
authors.lastname: Manning
mesh_terms: D000818:Animals; D016123:Campylobacter jejuni / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D002460:Cell Line; D002645:Chickens / Q000382:microbiology; D004251:DNA Transposable Elements; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000382:microbiology; D005700:Galactosyltransferases / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006801:Humans; D008070:Lipopolysaccharides / Q000096:biosynthesis*; D016254:Mutagenesis, Insertional; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D008070:Lipopolysaccharides; D037521:Virulence Factors; C023023:lipid-linked oligosaccharides; D005700:Galactosyltransferases