Translocation of Pseudomonas aeruginosa from the intestinal tract is mediated by the binding of ExoS to an Na,K-ATPase regulator, FXYD3.
The intestinal tract is considered the most important reservoir of Pseudomonas aeruginosa in intensive care units (ICUs). Gut colonization by P. aeruginosa underlies the development of invasive infections such as gut-derived sepsis. Intestinal colonization by P. aeruginosa is associated with higher ICU mortality rates. The translocation of endogenous P. aeruginosa from the colonized intestinal tract is an important pathogenic phenomenon. Here we identify bacterial and host proteins associated with bacterial penetration through the intestinal epithelial barrier. We first show by comparative genomic hybridization analysis that the exoS gene, encoding the type III effector protein, ExoS, was specifically detected in a clinical isolate that showed higher virulence in silkworms following midgut injection. We further show using a silkworm oral infection model that exoS is required both for virulence and for bacterial translocation from the midgut to the hemolymph. Using a bacterial two-hybrid screen, we show that the mammalian factor FXYD3, which colocalizes with and regulates the function of Na,K-ATPase, directly binds ExoS. A pulldown assay revealed that ExoS binds to the transmembrane domain of FXYD3, which also interacts with Na,K-ATPase. Na,K-ATPase controls the structure and barrier function of tight junctions in epithelial cells. Collectively, our results suggest that ExoS facilitates P. aeruginosa penetration through the intestinal epithelial barrier by binding to FXYD3 and thereby impairing the defense function of tight junctions against bacterial penetration.
Okuda
Jun
J
Department of Microbiology and Infection Control Science, Kyoto Pharmaceutical University, Yamashina, Kyoto, Japan.
Hayashi
Naoki
N
Okamoto
Masashi
M
Sawada
Shinji
S
Minagawa
Shu
S
Yano
Yoshitaka
Y
Gotoh
Naomasa
N
D036002:ADP Ribose Transferases / Q000378:metabolism*; D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D001692:Biological Transport; D012831:Bombyx / Q000382:microbiology*; D018938:Caco-2 Cells; D002460:Cell Line; D055028:Comparative Genomic Hybridization; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology*; D006458:Hemolymph / Q000382:microbiology*; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D008565:Membrane Proteins / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification / Q000378:metabolism / Q000472:pathogenicity* / Q000502:physiology; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism; D014774:Virulence
D001427:Bacterial Toxins; C091777:FXYD3 protein, human; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D036002:ADP Ribose Transferases; C444820:exoenzyme S; D000254:Sodium-Potassium-Exchanging ATPase
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