1. Fluorescent protein-based reporters reveal stress response of intracellular Salmonella enterica at level of single bacterial cells
- Author
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Michael Hensel, Katharina Olschewski, and Marc Schulte
- Subjects
Paraquat ,Salmonella typhimurium ,Immunology ,Biology ,Microbiology ,Flow cytometry ,03 medical and health sciences ,Mice ,Genes, Reporter ,Stress, Physiological ,Virology ,medicine ,Type III Secretion Systems ,Animals ,Humans ,Gene ,030304 developmental biology ,Cell Proliferation ,Polymyxin B ,0303 health sciences ,medicine.diagnostic_test ,030306 microbiology ,Intracellular parasite ,Macrophages ,Epithelial Cells ,Periplasmic space ,Gene Expression Regulation, Bacterial ,Hydrogen Peroxide ,biology.organism_classification ,Cell biology ,Dithiothreitol ,Luminescent Proteins ,RAW 264.7 Cells ,Salmonella enterica ,Cytoplasm ,Genes, Bacterial ,Single-Cell Analysis ,Intracellular ,Function (biology) ,HeLa Cells - Abstract
Intracellular bacteria such as Salmonella enterica are confronted with a broad array of defence mechanisms of their mammalian host cells. The ability to sense host cell-imposed damages, and to mount efficient stress responses are crucial for survival and proliferation of intracellular pathogens. The various combinations of host defence mechanisms acting on intracellular bacteria and their individual response also explain the occurrence of distinct subpopulations of intracellular S. enterica such as dormant or persisting, slowly or rapidly replicating cells. Here we describe a set of fluorescence protein (FP)-based reporter strains that were used to monitor the expression of cytoplasmic or periplasmic stress response systems of single bacterial cells. This is mediated by a fast-maturing FP as reporter for induction of stress response genes. We evaluated slower maturing FPs for a second function, that is, the analysis of the status of intracellular proliferation of pathogens. The combination of two FPs allows, at level of single bacterial cells, the interrogation of stress response and intracellular proliferation. Application of these reporters to S. enterica allowed us to detect and quantify distinct intracellular subpopulations with different levels of stress response and proliferation.
- Published
- 2020