Respiration and the F1Fo-ATPase Enhance Survival under Acidic Conditions in Escherichia coli

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Abstract

Besides amino acid decarboxylation, the ADP biosynthetic pathway was reported to enhance survival under extremely acidic conditions in Escherichia coli (Sun et al., J. Bacteriol. 193: 3072-3077, 2011). E. coli has two pathways for ATP synthesis from ADP: glycolysis and oxidative phosphorylation. We found in this study that the deletion of the F1Fo-ATPase, which catalyzes the synthesis of ATP from ADP and inorganic phosphate using the electro-chemical gradient of protons generated by respiration in E. coli, decreased the survival at pH 2.5. A mutant deficient in hemA encoding the glutamyl tRNA reductase, which synthesizes glutamate 1-semialdehyde also showed the decreased survival of E. coli at pH 2.5. Glutamate 1-semialdehyde is a precursor of heme synthesis that is an essential component of the respiratory chain. The ATP content decreased rapidly at pH 2.5 in these mutants as compared with that of their parent strain. The internal pH was lowered by the deletion of these genes at pH 2.5. These results suggest that respiration and the F1Fo-ATPase are still working at pH 2.5 to enhance the survival under such extremely acidic conditions. © 2012 Sun et al.

Figures

  • Table 1. Bacterial strains and plasmids used in this study.
  • Figure 1. Proton pumping activity of the mutants and the wild type strain. W3110 (wild type, parent strain of SE mutants), DK8, SE023 (atpE), and SE020 (atpD) were grown, and proton pumping activity was measured as described in Materials and Methods. ATP (1 mM) was added at zero time. doi:10.1371/journal.pone.0052577.g001
  • Figure 2. The survival of various mutants after 1 h challenge at pH 2.5. After W3110 (wild type, parent strain of SE mutants), SE020 (atpD), SE023 (atpE), DK8, SE022 (hemA), and SE021 (atpD hemA) had been grown in EG medium at pH 7.5 until OD600 reached 0.3 to 0.4, the cells were adapted for 4 h at pH 5.5 and challenged for 1 h at pH 2.5 as described in Materials and Methods. SE022 (hemA) and SE021 (atpD hemA) were precultured overnight with the addition of ALA (100 mg/ml) and then diluted with EG medium at pH7.5 without ALA. ALA was not added to media of pH 5.5 and 2.5. Data from three independent experiments are expressed as mean 6 S. D. Symbols: white bars, no addition; black bars, 0.1 mM glutamate was added to media of pH 5.5 and 2.5; gray bars, 0.1 mM arginine was added to media of pH 5.5 and 2.5; #, survival rate was too low to detect (less than 0.001%). The average values and standard deviations obtained from three experiments using separate cultures are represented. One asterisk, p,0.01 compared with the wild type; two asterisks, p,0.005 compared with the wild type. doi:10.1371/journal.pone.0052577.g002
  • Figure 3. ATP content of various mutants. DK8, SE020 (atpD), SE023 (atpE), SE022 (hemA), and W3110 (wild type, parent strain of SE mutants) were cultured as described in the legend of Fig. 2, and the ATP content was measured as described in Materials and Methods. Strains: 1, W3110 (wild type); 2, SE020 (atpD); 3, SE023 (atpE); 4, DK8; 5, SE022 (hemA). Data from three independent experiments are expressed as mean 6 S. D. #, the ATP content was less than 0.01 nmol/mg protein. The average values and standard deviations obtained from three experiments using separate cultures are represented. One asterisk, p,0.01 compared with the wild type; two asterisks, p,0.005 compared with the wild type. doi:10.1371/journal.pone.0052577.g003
  • Figure 4. Expression of the F1Fo-ATPase. DK8, W3110 (wild type), and SE023 (atpE) were grown at the pH indicated, and the amounts of F1 subunits were measured with Western blot analysis as described in Materials and Methods. M, molecular weight marker. doi:10.1371/journal.pone.0052577.g004
  • Table 2. Intracellular pH in various mutants.

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Sun, Y., Fukamachi, T., Saito, H., & Kobayashi, H. (2012). Respiration and the F1Fo-ATPase Enhance Survival under Acidic Conditions in Escherichia coli. PLoS ONE, 7(12). https://doi.org/10.1371/journal.pone.0052577

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