A Deep-Rough Mutant of
            <i>Campylobacter jejuni</i>
            81-176 Is Noninvasive for Intestinal Epithelial Cells

Kanipes, Margaret I.; Holder, Lindsay C.; Corcoran, Adrian T.; Moran, Anthony P.; Guerry, Patricia

doi:10.1128/iai.72.4.2452-2455.2004

Cited by 54 publications

(60 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The results of mutations to the C. jejuni 81-176 LOS length have varied; mutagenesis of cgtA lead to increased invasion of cell layers, whereas large truncation mutants had decreased invasion (50,51). In our study, adherence of the C. jejuni 11168 ΔcgtA/neuB was increased compared with wild type.…”

Section: Discussionmentioning

confidence: 48%

Glycan:glycan interactions: High affinity biomolecular interactions that can mediate binding of pathogenic bacteria to host cells

Day

Tran

Semchenko

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

101

104

View full text Add to dashboard Cite

Cells from all domains of life express glycan structures attached to lipids and proteins on their surface, called glycoconjugates. Cell-tocell contact mediated by glycan:glycan interactions have been considered to be low-affinity interactions that precede highaffinity protein-glycan or protein-protein interactions. In several pathogenic bacteria, truncation of surface glycans, lipooligosaccharide (LOS), or lipopolysaccharide (LPS) have been reported to significantly reduce bacterial adherence to host cells. Here, we show that the saccharide component of LOS/LPS have direct, high-affinity interactions with host glycans. Glycan microarrays reveal that LOS/LPS of four distinct bacterial pathogens bind to numerous host glycan structures. Surface plasmon resonance was used to determine the affinity of these interactions and revealed 66 high-affinity host-glycan:bacterial-glycan pairs with equilibrium dissociation constants (K D ) ranging between 100 nM and 50 μM. These glycan:glycan affinity values are similar to those reported for lectins or antibodies with glycans. Cell assays demonstrated that glycan:glycan interaction-mediated bacterial adherence could be competitively inhibited by either host cell or bacterial glycans. This is the first report to our knowledge of high affinity glycan:glycan interactions between bacterial pathogens and the host. The discovery of large numbers of glycan:glycan interactions between a diverse range of structures suggests that these interactions may be important in all biological systems.H ost surface glycosylation is ubiquitous and is targeted by pathogenic bacteria, viruses, fungi and parasites for adherence and toxin binding and by glycosidases (1). Escherichia coli type 1 fimbriae, FimH, is one of the most widely studied glycanrecognizing protein adhesins, with specificity for monomannose to oligomannose structures with the variability of the mannose structure bound leading to different tissue tropism (2). Other glycan-recognizing adhesins expressed by bacteria include the following: Pseudomonas aeruginosa lectins 1 and 2 (PA-IL and PA-IIL) that have specificity for galactose and fucose, respectively (3); Helicobacter pylori SabA, specific for sialic acid containing glycoconjugates including sialyLewis X; and BabAspecific for fucosylated glycoconjugates including Lewis B (4, 5). Although there are numerous known glycan binding adhesins, the adhesins of some bacteria that interact with host surface glycans remain unknown.Direct interactions between surface glycans (glycan:glycan interactions) have been reported in sea sponges as heterogenous glycan interactions, and in mouse embryo development and cancer where homodimers of Lewis X (LeX) or ganglioside structures play a role in cell adhesion and growth factor receptor interactions (6, 7). Outside of these reports, glycan:glycan interactions, when noted, have generally been considered to be low-affinity, weak interactions (8) that precede high-affinity protein:glycan or protein:protein interactions (1, 2, 5, 9).Interestingly, there...

show abstract

Section: Discussionmentioning

confidence: 48%

Glycan:glycan interactions: High affinity biomolecular interactions that can mediate binding of pathogenic bacteria to host cells

Day

Tran

Semchenko

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

101

104

View full text Add to dashboard Cite

show abstract

“…Absence of the LOS outer core is detrimental to C. jejuni growth and undermines outer membrane integrity. The inability of the NCTC11168⌬32-52 large deletion mutant to invade Caco-2 further supports a role for LOS in adherence and invasion (7,8,13).…”

mentioning

confidence: 99%

“…1), and modulation of LOS structure arises due to gene content diversity, allelic variation, and high-frequency sequence variation of homopolymeric tracts (10). The function of some genes in the cluster has been elucidated, and individual mutations in waaC and waaF have yielded deep rough mutants (7,8,16). Here, we describe the construction and phenotype of a mutant in which the core LOS biosynthesis gene cluster has been removed.…”

mentioning

confidence: 99%

Creation of a Large Deletion Mutant ofCampylobacter jejuniReveals that the Lipooligosaccharide Gene Cluster Is Not Required for Viability

Marsden

Everest

et al. 2009

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…The carbohydrate structure of the outer core region of C. jejuni 81-176 has been shown to mimic primarily GM 2 ( Fig. 1) and GM 3 (12,17) human gangliosides, although minor forms of GD 2 and GD 1b ganglioside mimics have been reported (12). The variation between GM 2 and GM 3 mimicry is due to the presence of a phase-variable homopolymeric G tract in the cgtA gene, which encodes an N-acetylgalactosaminyltransferase (12).…”

mentioning

confidence: 99%

Mutation ofwaaC, Encoding Heptosyltransferase I inCampylobacter jejuni81-176, Affects the Structure of both Lipooligosaccharide and Capsular Carbohydrate

et al. 2006

Self Cite

View full text Add to dashboard Cite

Campylobacter jejuni 81-176 lipooligosaccharide (LOS) is composed of two covalently linked domains: lipid A, a hydrophobic anchor, and a nonrepeating core oligosaccharide, consisting of an inner and outer core region. We report the isolation and characterization of the deepest rough C. jejuni 81-176 mutant by insertional mutagenesis into the waaC gene, encoding heptosyltransferase I that catalyzes the transfer of the first L-glycero-D-manno-heptose residue to 3-deoxy-D-manno-octulosonic residue (Kdo)-lipid A. Tricine gel electrophoresis, followed by silver staining, showed that site-specific mutation in the waaC gene resulted in the expression of a severely truncated LOS compared to wild-type strain 81-176. Gas-liquid chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy showed that the waaC LOS species lacked all sugars distal to Kdo-lipid A. Parallel structural studies of the capsular polysaccharides of the wild-type strain 81-176 and waaC mutant revealed loss of the 3-O-methyl group in the waaC mutant. Complementation of the C. jejuni mutant by insertion of the wild-type C. jejuni waaC gene into a chromosomal locus resulted in LOS and capsular structures identical to those expressed in the parent strain. We also report here the presence of O-methyl phosphoramidate in wild-type strain 81-176 capsular polysaccharide.

show abstract

A Deep-Rough Mutant of Campylobacter jejuni 81-176 Is Noninvasive for Intestinal Epithelial Cells

Abstract: A waaF mutant of Campylobacter jejuni 81-176 showed decreased invasion of INT407 cells in vitro and increased sensitivity to some antibiotics compared to what was seen with the wild-type strain.

Cited by 54 publications

References 21 publications

Glycan:glycan interactions: High affinity biomolecular interactions that can mediate binding of pathogenic bacteria to host cells

Glycan:glycan interactions: High affinity biomolecular interactions that can mediate binding of pathogenic bacteria to host cells

Creation of a Large Deletion Mutant ofCampylobacter jejuniReveals that the Lipooligosaccharide Gene Cluster Is Not Required for Viability

Mutation ofwaaC, Encoding Heptosyltransferase I inCampylobacter jejuni81-176, Affects the Structure of both Lipooligosaccharide and Capsular Carbohydrate

Contact Info

Product

Resources

About