The basolateral vesicle sorting machinery and basolateral proteins are recruited to the site of enteropathogenic E. coli microcolony growth at the apical membrane
| Autor: | Maj Ulrichsen, Lene N. Nejsum, Gitte Pedersen, Hans Nymand Pedersen, Helene Halkjær Jensen, Manuel R. Amieva, Anne-Sofie B. Schelde, Charlotte Toft, Frédéric H. Login |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cell Membranes lcsh:Medicine Bacterial Adhesion Madin Darby Canine Kidney Cells Enteropathogenic Escherichia coli Fluorescence Microscopy Cell polarity lcsh:Science Escherichia coli Infections Epithelial polarity Staining Microscopy Multidisciplinary Escherichia coli Proteins Cell Staining Light Microscopy Cell Polarity Basolateral plasma membrane Cell biology Transport protein Protein Transport Cellular Structures and Organelles Junctional Complexes Research Article trans-Golgi Network Cell Physiology Exocyst Biology Research and Analysis Methods Microbiology Tight Junctions 03 medical and health sciences Dogs Virology Journal Article Animals Vesicles 030102 biochemistry & molecular biology Host Cells lcsh:R Cell Membrane Biology and Life Sciences Membrane Proteins Cell Biology Apical membrane biochemical phenomena metabolism and nutrition 030104 developmental biology Specimen Preparation and Treatment Host cell plasma membrane bacteria lcsh:Q Rab Viral Transmission and Infection |
| Zdroj: | Pedersen, G A, Jensen, H H, Schelde, A-S B, Toft, C, Pedersen, H N, Ulrichsen, M, Login, F H, Amieva, M R & Nejsum, L N 2017, ' The basolateral vesicle sorting machinery and basolateral proteins are recruited to the site of enteropathogenic E. coli microcolony growth at the apical membrane ', P L o S One, vol. 12, no. 6, e0179122 . https://doi.org/10.1371/journal.pone.0179122 PLoS ONE, Vol 12, Iss 6, p e0179122 (2017) PLoS ONE Aalborg University |
| DOI: | 10.1371/journal.pone.0179122 |
| Popis: | Foodborne Enteropathogenic Escherichia coli (EPEC) infections of the small intestine cause diarrhea especially in children and are a major cause of childhood death in developing countries. EPEC infects the apical membrane of the epithelium of the small intestine by attaching, effacing the microvilli under the bacteria and then forming microcolonies on the cell surface. We first asked the question where on epithelial cells EPEC attaches and grows. Using models of polarized epithelial monolayers, we evaluated the sites of initial EPEC attachment to the apical membrane and found that EPEC preferentially attached over the cell-cell junctions and formed microcolonies preferentially where three cells come together at tricellular tight junctions. The ability of EPEC to adhere increased when host cell polarity was compromised yielding EPEC access to basolateral proteins. EPEC pedestals contain basolateral cytoskeletal proteins. Thus, we asked if attached EPEC causes reorganization the protein composition of the host cell plasma membrane at sites of microcolony formation. We found that EPEC microcolony growth at the apical membrane resulted in a local accumulation of basolateral plasma membrane proteins surrounding the microcolony. Basolateral marker protein aquaporin-3 localized to forming EPEC microcolonies. Components of the basolateral vesicle targeting machinery were re-routed. The Exocyst (Exo70) was recruited to individual EPEC as was the basolateral vesicle SNARE VAMP-3. Moreover, several Rab variants were also recruited to the infection site, and their dominant-negative equivalents were not. To quantitatively study the recruitment of basolateral proteins, we created a pulse of the temperature sensitive basolateral VSVG, VSVG3-SP-GFP, from the trans-Golgi Network. We found that after release from the TGN, significantly more VSVG3-SP-GFP accumulated at the site of microcolony growth than on equivalent membrane regions of uninfected cells. This suggests that trafficking of vesicles destined for the basolateral membrane are redirected to the apical site of microcolony growth. Thus, in addition to disrupting host cell fence function, local host cell plasma membrane protein composition is changed by altered protein trafficking and recruitment of basolateral proteins to the apical microcolony. This may aid EPEC attachment and subsequent microcolony growth. |
| Databáze: | OpenAIRE |
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