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. 2019 Nov 28;2:440. doi: 10.1038/s42003-019-0686-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 — Dimerization of ToxT at the helix α3 interface is required for DNA binding. a Model of the ToxT dimer interface. b LexA-fusion bacterial two-hybrid dimerization assay of ToxT_EPI dimer interface mutants. Error bars indicate standard deviation. A western blot of the LexA-ToxT_EPI fusions confirming expression is shown in Supplementary Fig. 4. c Electrophoretic mobility shift assay of wild-type and K158E ToxT_ENV256 binding to the tcpA promoter. All lanes contain 9 nM DIG-labeled 84 bp segment of dsDNA containing the ToxT-binding sites from the tcpA promoter. Lane 1, free DNA; lane 2, 0.098 μg wild-type ToxT_ENV256; lane 3, 0.195 μM wild-type ToxT_ENV256; lane 4, 0.39 μM wild-type ToxT_ENV256; lane 5, 0.78 μM wild-type ToxT_ENV256; lane 6, 1.56 μM wild-type ToxT_ENV256; 7, free DNA; lane 8, 0.098 μM K158E ToxT_ENV256; lane 9, 0.195 μM K158E ToxT_ENV256; lane 10, 0.39 μM K158E ToxT_ENV256; lane 11, 0.78 μM K158E ToxT_ENV256; lane 12, 1.56 μM K158E ToxT_ENV256. Error bars are indicated (n = 3 experiments). A western blot of the LexAToxTEPI fusions confirming expression of protein has been provided as Supplementary Fig. 4.