Fluorescent protein-based reporters reveal stress response of intracellular Salmonella enterica at level of single bacterial cells
|antimicrobial activity; Cell Biology; containing vacuole; cytosolic stress response; EFFECTORS; ENDOSOMAL SYSTEM; flow cytometry; fluorescent proteins; III SECRETION SYSTEM; intracellular pathogen; Microbiology; PATHOGENICITY ISLAND-2; periplasmic stress response; PERSISTERS; PROTECTION; REDUCTASE; Salmonella‐ stress response; SULFOXIDE; TYPHIMURIUM; VIRULENCE
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.
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checked on Feb 22, 2024