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. 2020 Nov 30;10:20837. doi: 10.1038/s41598-020-77738-z

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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The kinase activity of RsbT is essential for SigB activation with RsbR1 paralogues antagonising such activity in unstressed and stressed L. monocytogenes. (A) Expression of the P_lmo2230::egfp reporter fusion as proxy of SigB activity in the indicated strains in no stress and stress (30 min, 0.5 M NaCl) conditions. Shown is the fluorescence intensity registered in the bacterial population by flow cytometry. (B) Side-to-side comparison of SigB activity in wild type compared to the mutant lacking the first phosphorylatable threonine residue, T175, and the Δ4 mutant lacking functional RsbR1 paralogues. Data shown in panels (A,B) are representative of a total of six samples (3 biological replicates with two technical replicates each). (C) Mean fluorescence values of the expression data depicted in panel A. Data shown are the mean and standard deviation of a total of six samples, corresponding to three biological replicates with two technical replicates each. ***, P ≤ 0.005; n. s., not significant (one-way ANOVA with Bonferroni’s multiple comparisons test).