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. 2018 Mar 7;9(3):381. doi: 10.1038/s41419-018-0423-2

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© The Author(s) 2018

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. 3 — a A diagram of the enzymes (genes) and metabolites of the TCA cycle and the alternative metabolic path of succinate through YgfH. b The survival of N2 worms fed with the wild-type EHEC strain EDL933 (EDL933) and the isogenic EDL933 strains with deletion of gltA (EDL933:ΔgltA), acnB (EDL933:ΔacnB), and sdhA (EDL933:ΔsdhA) were examined. Deletions of gltA (median N2 lifespan = 8.5 ± 0.7 days, P < 0.0001), acnB (median N2 lifespan = 8.0 ± 0.1 days, P < 0.0001), and sdhA (median N2 lifespan = 12.0 ± 2.2 days, P < 0.0001) all conferred the attenuated toxic phenotype compared to the wild-type EDL933 (the median N2 lifespan = 6.0 ± 0.1 days). Moreover, EDL933:ΔsdhA mutant was significantly less toxic compared to the EDL933:ΔgltA (P < 0.0001) and EDL933:ΔacnB (P < 0.0001). c The survival of N2 worms fed with the wild-type EDL933 and the isogenic deletion strains of icdA (EDL933:ΔicdA), sucAB (EDL933:ΔsucAB), and sdhA (EDL933:ΔsdhA) were examined. Deletions of icdA (median N2 lifespan = 9.0 ± 1.4 days, P < 0.005), sucAB (median N2 lifespan = 13.5 ± 2.1 days, P < 0.0001), and sdhA (median N2 lifespan = 13.5 ± 2.1 days, P < 0.0001) all conferred the attenuated toxic phenotype compared to the wild-type EDL933 (the median N2 lifespan = 6 ± 0.2 days). The EDL933:ΔsdhA mutant was significantly less toxic compared to both the EDL933:ΔicdA (P < 0.0001) and the EDL933:ΔsucAB mutant (P < 0.05). d The survival of N2 worms fed with the wild-type EDL933 and the isogenic deletion strains of sucCD (EDL933:ΔsucCD) and sdhA (EDL933:ΔsdhA) were examined. Deletion of sucCD (median N2 lifespan = 9.0 ± 1.0 days, P < 0.0001) and sdhA (median N2 lifespan = 11.3 ± 1.5 days, P < 0.0001) conferred the attenuated toxic phenotype compared to the wild-type EDL933 (the median N2 lifespan = 6.3 ± 0.5 days). Moreover, EDL933:ΔsdhA mutant was significantly less toxic compared to the EDL933:ΔsucCD (P < 0.0001). e The survival of N2 worms fed with the wild-type EDL933 and the isogenic deletion strains of fumCA (EDL933:ΔfumCA), mdh (EDL933:Δmdh), and sdhA (EDL933:ΔsdhA) were examined. Deletions of fumCA (median N2 lifespan = 9.0 ± 0.1 days, P < 0.0001), mdh (median N2 lifespan = 8.5 ± 0.5 days, P < 0.0001), and sdhA (median N2 lifespan = 10.5 ± 0.5 days, P < 0.0001) all conferred the attenuated toxic phenotype compared to the wild-type EDL933 (median N2 lifespan = 6.1 ± 0.1 days). Moreover, EDL933:ΔsdhA mutant was significantly less toxic compared to the EDL933:ΔfumCA (P < 0.0001) and EDL933:Δmdh (P < 0.0001). f The survival of N2 worms fed with the wild-type EDL933 and the isogenic deletion strains of ygfH (EDL933:ΔygfH), sdhA (EDL933:ΔsdhA), and the ygfH and sdhA double mutant (EDL933:ΔygfHΔsdhA) were examined. Deletion of ygfH (median N2 lifespan = 6.3 ± 0.4 days, P = 0.102) did not attenuate its toxicity. Although deletion of the ygfH and sdhA double mutant (EDL933:ΔygfHΔsdhA, median N2 lifespan = 11.5 ± 0.7 days, P < 0.0001) conferred the attenuated toxic phenotype compared to the wild-type EDL933 (the median N2 lifespan = 5.0 ± 0.5 days), the virulence of EDL933:ΔygfHΔsdhA strain was similar to the EDL933:Δ sdhA mutant (median N2 lifespan = 12.5 ± 0.7 days, P = 0.691)