Physical and antibiotic stresses require activation of the RsbU phosphatase to induce the general stress response in Listeria monocytogenes.
Among pathogenic strains of Listeria monocytogenes, the sigma(B) transcription factor has a pivotal role in the outcome of food-borne infections. This factor is activated by diverse stresses to provide general protection against multiple challenges, including those encountered during gastrointestinal passage. It also acts with the PrfA regulator to control virulence genes needed for entry into intestinal lumen cells. Environmental and nutritional signals modulate sigma(B) activity via a network that operates by the partner switching mechanism, in which protein interactions are controlled by serine phosphorylation. This network is well characterized in the related bacterium Bacillus subtilis. A key difference in Listeria is the presence of only one input phosphatase, RsbU, instead of the two found in B. subtilis. Here, we aim to determine whether this sole phosphatase is required to convey physical, antibiotic and nutritional stress signals, or if additional pathways might exist. To that end, we constructed L. monocytogenes 10403S strains bearing single-copy, sigma(B)-dependent opuCA-lacZ reporter fusions to determine the effects of an rsbU deletion under physiological conditions. All stresses tested, including acid, antibiotic, cold, ethanol, heat, osmotic and nutritional challenge, required RsbU to activate sigma(B). This was of particular significance for cold stress activation, which occurs via a phosphatase-independent mechanism in B. subtilis. We also assayed the effects of the D80N substitution in the upstream RsbT regulator that activates RsbU. The mutant had a phenotype consistent with low and uninducible phosphatase activity, but nonetheless responded to nutritional stress. We infer that RsbU activity but not its induction is required for nutritional signalling, which would enter the network downstream from RsbU.
Shin
Ji-Hyun
JH
Department of Food Science and Technology, University of California, Davis, CA 95616, USA.
Brody
Margaret S
MS
Department of Food Science and Technology, University of California, Davis, CA 95616, USA.
Price
Chester W
CW
Department of Food Science and Technology, University of California, Davis, CA 95616, USA.
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003080:Cold Temperature; D004789:Enzyme Activation; D015964:Gene Expression Regulation, Bacterial; D006358:Hot Temperature; D008089:Listeria monocytogenes / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000502:physiology*; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D013312:Stress, Physiological
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D010744:Phosphoric Monoester Hydrolases