Imaging InlC Secretion to Investigate Cellular Infection by the Bacterial Pathogen Listeria monocytogenes |
| |
Authors: | Andreas Kühbacher Edith Gouin Pascale Cossart Javier Pizarro-Cerdá |
| |
Institution: | 1.Unité des Interactions Bactéries Cellules, Pasteur Institute;2.INSERM U604;3.Institut National de la Recherche Agronomique (INRA), USC2020 |
| |
Abstract: | Bacterial intracellular pathogens can be conceived as molecular tools to dissect cellular signaling cascades due to their capacity to exquisitely manipulate and subvert cell functions which are required for the infection of host target tissues. Among these bacterial pathogens, Listeria monocytogenes is a Gram positive microorganism that has been used as a paradigm for intracellular parasitism in the characterization of cellular immune responses, and which has played instrumental roles in the discovery of molecular pathways controlling cytoskeletal and membrane trafficking dynamics. In this article, we describe a robust microscopical assay for the detection of late cellular infection stages of L. monocytogenes based on the fluorescent labeling of InlC, a secreted bacterial protein which accumulates in the cytoplasm of infected cells; this assay can be coupled to automated high-throughput small interfering RNA screens in order to characterize cellular signaling pathways involved in the up- or down-regulation of infection. |
| |
Keywords: | Immunology Issue 79 HeLa Cells Listeria monocytogenes Gram-positive Bacterial Infections Fluorescence High-Throughput Screening Assays RNA Interference Listeria monocytogenes Infection microscopy small interfering RNA |
|