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. 2019 Nov 21;10:5288. doi: 10.1038/s41467-019-13296-x

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© The Author(s) 2019

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — MpaR functions as an anti-repressor of both monocin and ϕ10403S. a A plaque forming assay of WT Lm, ΔmpaR and Δmon mutants, as well as their complemented strains harboring mpaR gene on pPL2 (ΔmpaR+pPL2-mpaR and Δmon+pPL2-mpaR, respectively), 4 h post MC treatment at 30˚C. Also included ∆mpaR mutant that was introduced with mpaR-H54A variant on pPL2 (ΔmpaR+pPL2-mpaR-H54A). The results are normalized to PFU of WT bacteria. Error bars represent standard deviation of 3 independent experiments. b qRT-PCR analysis of transcription levels of indicated genes of monocin-cluster or ϕ10403S under SOS (4 h post MC treatment at 30˚C) in WT and ΔmpaR bacteria. Transcription levels are represented as relative quantity (RQ), relative to their levels in WT bacteria. The data represent three independent experiments. Error bars indicate a 95% confidence interval. c Growth of WT Lm, ΔmpaR and Δmon, as well as their complemented strains harboring the mpaR gene on pPL2 under the constitutive veg promoter (ΔmpaR+pPL2-mpaR and Δmon+pPL2-mpaR, respectively) without (left panel) and with (right panel) MC. The data shows the mean and the standard deviation of three independent biological repeats. d Western blot analysis of CI-like repressors cleavage by MpaR. Δmon/Δϕ bacteria harboring pPL2 expressing translational fusions of mon-CI-like-GFP (left panel) and ϕ10403S CI-like-6-His (right panel), in addition to MpaR or MpaR-H54A variant, under the tetR promoter. Equal amounts of total protein from bacteria grown in the presence and absence of MC were separated on 15% SDS-PAGE, blotted and probed with anti-GFP antibodies for mon-CI-like repressor or anti-6His antibodies for ϕ10403S-CI-like repressor. The experiment was performed 3 times, and the figure shows representative blots. Source data are provided as a Source Data file.