Campylobacter jejuni enters gut epithelial cells and impairs intestinal barrier function through cleavage of occludin by serine protease HtrA

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Details zur Publikation

Autorinnen und Autoren: Harrer A, Buecker R, Böhm M, Zarzecka U, Tegtmeyer N, Sticht H, Schulzke JD, Backert S
Zeitschrift: Gut Pathogens
Jahr der Veröffentlichung: 2019
Band: 11
ISSN: 1757-4749


Abstract

Campylobacter jejuni secretes HtrA (high temperature requirement protein A), a serine protease that is involved in virulence. Here, we investigated the interaction of HtrA with the host protein occludin, a tight junction strand component. Immunofluorescence studies demonstrated that infection of polarized intestinal Caco-2 cells with C. jejuni strain 81-176 resulted in a redistribution of occludin away from the tight junctions into the cytoplasm, an effect that was also observed in human biopsies during acute campylobacteriosis. Occludin knockout Caco-2 cells were generated by CRISPR/Cas9 technology. Inactivation of this gene affected the polarization of the cells in monolayers and transepithelial electrical resistance (TER) was reduced, compared to wild-type Caco-2 cells. Although tight junctions were still being formed, occludin deficiency resulted in a slight decrease of the tight junction plaque protein ZO-1, which was redistributed off the tight junction into the lateral plasma membrane. Adherence of C. jejuni to Caco-2 cell monolayers was similar between the occludin knockout compared to wild-type cells, but invasion was enhanced, indicating that deletion of occludin allowed larger numbers of bacteria to pass the tight junctions and to reach basal membranes to target the fibronectin receptor followed by cell entry. Finally, we discovered that purified C. jejuni HtrA cleaves recombinant occludin in vitro to release a 37 kDa carboxy-terminal fragment. The same cleavage fragment was observed in Western blots upon infection of polarized Caco-2 cells with wild-type C. jejuni, but not with isogenic Delta htrA mutants. HtrA cleavage was mapped to the second extracellular loop of occludin, and a putative cleavage site was identified. In conclusion, HtrA functions as a secreted protease targeting the tight junctions, which enables the bacteria by cleaving occludin and subcellular redistribution of other tight junction proteins to transmigrate using a paracellular mechanism and subsequently invade epithelial cells.


FAU-Autorinnen und Autoren / FAU-Herausgeberinnen und Herausgeber

Backert, Steffen Prof. Dr.
Lehrstuhl für Mikrobiologie
Böhm, Manja Dr.
Lehrstuhl für Mikrobiologie
Harrer, Aileen
Lehrstuhl für Mikrobiologie
Sticht, Heinrich Prof. Dr.
Professur für Bioinformatik
Zarzecka, Urszula
Lehrstuhl für Mikrobiologie


Einrichtungen weiterer Autorinnen und Autoren

Charité - Universitätsmedizin Berlin


Zitierweisen

APA:
Harrer, A., Buecker, R., Böhm, M., Zarzecka, U., Tegtmeyer, N., Sticht, H.,... Backert, S. (2019). Campylobacter jejuni enters gut epithelial cells and impairs intestinal barrier function through cleavage of occludin by serine protease HtrA. Gut Pathogens, 11. https://dx.doi.org/10.1186/s13099-019-0283-z

MLA:
Harrer, Aileen, et al. "Campylobacter jejuni enters gut epithelial cells and impairs intestinal barrier function through cleavage of occludin by serine protease HtrA." Gut Pathogens 11 (2019).

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Zuletzt aktualisiert 2019-06-03 um 13:08