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000604260 1001_ $$0P:(DE-H253)PIP1087856$$aThuenauer, Roland$$b0
000604260 245__ $$aThe Lectin LecB Induces Patches with Basolateral Characteristics at the Apical Membrane to Promote Pseudomonas aeruginosa Host Cell Invasion
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000604260 520__ $$aThe opportunistic bacterium Pseudomonas aeruginosa can infect mucosal tissues of the human body. To persist at the mucosal barrier, this highly adaptable pathogen has evolved many strategies, including invasion of host cells. Here, we show that the P. aeruginosa lectin LecB binds and cross-links fucosylated receptors at the apical plasma membrane of epithelial cells. This triggers a signaling cascade via Src kinases and phosphoinositide 3-kinase (PI3K), leading to the formation of patches enriched with the basolateral marker phosphatidylinositol (3,4,5)-trisphosphate (PIP3) at the apical plasma membrane. This identifies LecB as a causative bacterial factor for activating this well-known host cell response that is elicited upon apical binding of P. aeruginosa. Downstream from PI3K, Rac1 is activated to cause actin rearrangement and the outgrowth of protrusions at the apical plasma membrane. LecB-triggered PI3K activation also results in aberrant recruitment of caveolin-1 to the apical domain. In addition, we reveal a positive feedback loop between PI3K activation and apical caveolin-1 recruitment, which provides a mechanistic explanation for the previously observed implication of caveolin-1 in P. aeruginosa host cell invasion. Interestingly, LecB treatment also reversibly removes primary cilia. To directly prove the role of LecB for bacterial uptake, we coated bacterium-sized beads with LecB, which drastically enhanced their endocytosis. Furthermore, LecB deletion and LecB inhibition with l-fucose diminished the invasion efficiency of P. aeruginosa bacteria. Taken together, the results of our study identify LecB as a missing link that can explain how PI3K signaling and caveolin-1 recruitment are triggered to facilitate invasion of epithelial cells from the apical side by P. aeruginosa. IMPORTANCE An intriguing feature of the bacterium P. aeruginosa is its ability to colonize highly diverse niches. P. aeruginosa can, besides forming biofilms, also enter and proliferate within epithelial host cells. Moreover, research during recent years has shown that P. aeruginosa possesses many different mechanisms to invade host cells. In this study, we identify LecB as a novel invasion factor. In particular, we show that LecB activates PI3K signaling, which is connected via a positive feedback loop to apical caveolin-1 recruitment and leads to actin rearrangement at the apical plasma membrane. This provides a unifying explanation for the previously reported implication of PI3K and caveolin-1 in host cell invasion by P. aeruginosa. In addition, our study adds a further function to the remarkable repertoire of the lectin LecB, which is all brought about by the capability of LecB to recognize fucosylated glycans on many different niche-specific host cell receptors. 
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000604260 7001_ $$0P:(DE-HGF)0$$aKühn, Katja$$b1$$eCorresponding author
000604260 7001_ $$0P:(DE-HGF)0$$aGuo, Yubing$$b2
000604260 7001_ $$0P:(DE-HGF)0$$aKotsis, Fruzsina$$b3
000604260 7001_ $$0P:(DE-HGF)0$$aXu, Maokai$$b4
000604260 7001_ $$0P:(DE-HGF)0$$aTrefzer, Anne$$b5
000604260 7001_ $$0P:(DE-HGF)0$$aAltmann, Silke$$b6
000604260 7001_ $$0P:(DE-HGF)0$$aWehrum, Sarah$$b7
000604260 7001_ $$0P:(DE-HGF)0$$aHeshmatpour, Najmeh$$b8
000604260 7001_ $$0P:(DE-HGF)0$$aFaust, Brian$$b9
000604260 7001_ $$0P:(DE-HGF)0$$aLandi, Alessia$$b10
000604260 7001_ $$0P:(DE-HGF)0$$aDiedrich, Britta$$b11
000604260 7001_ $$0P:(DE-HGF)0$$aDengjel, Jörn$$b12
000604260 7001_ $$0P:(DE-HGF)0$$aKuehn, Stefan$$b13$$eCorresponding author
000604260 7001_ $$0P:(DE-HGF)0$$aImberty, Anne$$b14
000604260 7001_ $$0P:(DE-HGF)0$$aRömer, Winfried$$b15
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