Several pathogenic bacteria, including Listeria monocytogenes, use an F-actin-dependent motility process to spread between mammalian cells. Actin ‘comet tails’ propel Lm through the cytoplasm, resulting in bacteria-containing membrane protrusions that are internalized by neighboring cells. The mechanism by which L. monocytogenes overcomes cortical membrane tension to generate protrusions is unknown. In this work, I identify bacterial and host proteins that directly regulate the formation of protrusions. First, I show that efficient cell-cell spread in polarized epithelial cells requires the secreted Lm virulence protein, InlC. I next identify the mammalian adaptor protein Tuba as a ligand of InlC. InlC binds to a C-terminal SH3 domain in Tuba, which normally engages the human actin regulatory protein N-WASP. InlC promotes protrusion formation by inhibiting Tuba and N-WASP function, most likely by impairing binding of N-WASP to the Tuba SH3 domain. Tuba and N-WASP are known to control the structure of apical
junctions in epithelial cells [1]. I demonstrate that, by inhibiting Tuba and N-WASP, InlC transforms taut apical cell-cell junctions into structures with a “slack” morphology. Experiments with Myosin II inhibitors indicate that InlC-mediated perturbation of cell junctions accounts for the role of this bacterial protein in protrusion formation. Collectively, my results suggest that InlC enhances bacterial dissemination by relieving cortical tension in apical junctions, thereby enhancing the ability of motile bacteria to deform the plasma membrane into protrusions to allow their spread into neighbouring cells.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:OTU.1807/19075 |
| Date | 19 February 2010 |
| Creators | Rajabian, Tina |
| Contributors | Gray-Owen, Scott |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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