FlaC, a protein of <i>Campylobacter jejuni</i> TGH9011 (ATCC43431) secreted through the flagellar apparatus, binds epithelial cells and influences cell invasion

Song, Young Chan; Jin, Sun; Louie, Helena; Ng, David; Lau, Richard R.; Zhang, Y.; Weerasekera, Ranjuna; Rashid, Sabih; Ward, Lucy A.; Der, Sandy D.; Chan, Vivien

doi:10.1111/j.1365-2958.2004.04175.x

Cited by 133 publications

(107 citation statements)

References 47 publications

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“…The recent emergence of antibiotic-resistant strains has triggered renewed interest for the understanding of the pathogenesis of C. jejuni (2,3), which could lead to the identification of novel targets that could be further investigated for the development of therapeutics. A variety of virulence factors has been identified, such as the lipo-oligosaccharide (4 -7), capsule (8), flagellum (9), toxins (10), and adhesins (11)(12)(13)(14). They allow for colonization and invasion of the intestinal epithelium and protection against host immune defense systems.…”

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confidence: 99%

Cj1121c, a Novel UDP-4-keto-6-deoxy-GlcNAc C-4 Aminotransferase Essential for Protein Glycosylation and Virulence in Campylobacter jejuni

Somalinga

Merkx-Jacques

Ratnayake

et al. 2006

Journal of Biological Chemistry

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Campylobacter jejuni produces glycoproteins that are essential for virulence. These glycoproteins carry diacetamidobacillosamine (DAB), a sugar that is not found in humans. Hence, the enzymes responsible for DAB synthesis represent potential therapeutic targets. We describe the biochemical characterization of Cj1121c, a putative aminotransferase encoded by the general protein glycosylation locus, to assess its role in DAB biosynthesis. By using overexpressed and affinity-purified enzyme, we demonstrate that Cj1121c has pyridoxal phosphate-and glutamate-dependent UDP-4-keto-6-deoxy-GlcNAc C-4 transaminase activity and produces UDP-4-amino-4,6-dideoxy-GlcNAc. This is consistent with a role in DAB biosynthesis and distinguishes Cj1121c from Cj1294, a homologous UDP-2-acetamido-2,6-dideoxy-␤-L-arabino-4-hexulose C-4 aminotransferase that we characterized previously. We show that Cj1121c can also use this 4-keto-arabino sugar indirectly as a substrate, that Cj1121c and Cj1294 are active simultaneously in C. jejuni, and that the activity of Cj1121c is preponderant under standard growth conditions. Kinetic data indicate that Cj1121c has a slightly higher catalytic efficiency than Cj1294 with regard to the 4-keto-arabino substrate. By site-directed mutagenesis, we show that residues Glu-158 and Leu-131 are not essential for catalysis or for substrate specificity contrary to expectations. We further demonstrate that a cj1121c knock-out mutant is impaired for flagella-mediated motility, for invasion of intestinal epithelial cells, and for persistence in the chicken intestine, clearly demonstrating that Cj1121c is essential for host colonization and virulence. Finally, we show that cj1121c is necessary for protein glycosylation by lectin Western blotting. Collectively, these results validate Cj1121c as a promising drug target and provide the means to assay for inhibitors.

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confidence: 99%

Cj1121c, a Novel UDP-4-keto-6-deoxy-GlcNAc C-4 Aminotransferase Essential for Protein Glycosylation and Virulence in Campylobacter jejuni

Somalinga

Merkx-Jacques

Ratnayake

et al. 2006

Journal of Biological Chemistry

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“…Three prototype C. jejuni strains were used, to represent a variety of potential virulence mechanisms. NCTC11168, the first sequenced C. jejuni strain (1); the invasive strain, ATCC81-176 (3); TGH9011 (ATCC43431), a clinical isolate characterized at the University of Toronto, and a noninvasive flgF isogenic mutant of TGH9011 (17). All strains were incubated in Mueller-Hinton broth and regrown on Columbia-blood-agar plates under microaerophilic conditions (48 h; 37°C).…”

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confidence: 99%

Campylobacter jejuni Mediated Disruption of Polarized Epithelial Monolayers is Cell-Type Specific, Time Dependent, and Correlates With Bacterial Invasion

2008

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The precise mechanism by which the most common cause of bacterial enterocolitis in humans, Campylobacter jejuni, perturbs the intestinal mucosa remains elusive. To define effects of C. jejuni infection on mucosal permeability, Madin-Darby canine kidney (MDCK)-I and T84 cell monolayers were infected with C. jejuni for up to 48 h. All three tested C. jejuni strains caused a 73-78% reduction in transepithelial electrical resistance (TER) in intestinal (T84) cell monolayers, whereas only one strain slightly reduced TER of MDCK-I cells by 25% after 48 h infection. Infection with C. jejuni strains also caused a 2.3-4.5-fold increase in dextran permeability, but only in T84 cells. C. jejuni infection of monolayers also caused morphologic changes in desmosomes, observed by transmission electron microscopy. The cell-type specificity, demonstrated by increased T84 monolayer permeability, correlated with higher bacterial invasion into these cells, relative to MDCK-I cells. In T84 cells, invasion and bacterial translocation preceded barrier disruption and inhibition of C. jejuni invasion using a pharmacological inhibitor of phosphoinositide 3-kinase, reduced the drop in TER. These findings suggest that C. jejuni disruption of monolayers is mediated by invasion, provide new insights into C. jejuni-host epithelial barrier interactions, and offer potential mechanisms of intestinal injury and chronic immune stimulation. (Pediatr Res 64: 599-604, 2008)

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“…In Campyrobacter jejuni TGH9011, a Gram-negative spiral bacterium that perhaps lacks a type-III secretion apparatus, flagellin is excreted from the cells and is used for the recognition of host cells. 20) On the other hand, in strain A1, flagellin localizes to the cell surface and functions as a receptor for alginate and a regulator of the structure and function of the cell surface, both of which influence cell viability in the alginate medium. A question will be raised as to why flagellin-like protein shows specific and extremely high alginate-binding ability.…”

Section: Genome Sequencementioning

confidence: 99%

Superchannel of Bacteria: Biological Significance and New Horizons

Murata

Kawai

Mikami

et al. 2008

Bioscience, Biotechnology, and Biochemistry

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FlaC, a protein of Campylobacter jejuni TGH9011 (ATCC43431) secreted through the flagellar apparatus, binds epithelial cells and influences cell invasion

Cited by 133 publications

References 47 publications

Cj1121c, a Novel UDP-4-keto-6-deoxy-GlcNAc C-4 Aminotransferase Essential for Protein Glycosylation and Virulence in Campylobacter jejuni

Cj1121c, a Novel UDP-4-keto-6-deoxy-GlcNAc C-4 Aminotransferase Essential for Protein Glycosylation and Virulence in Campylobacter jejuni

Campylobacter jejuni Mediated Disruption of Polarized Epithelial Monolayers is Cell-Type Specific, Time Dependent, and Correlates With Bacterial Invasion

Superchannel of Bacteria: Biological Significance and New Horizons

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