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Cytoplasmic acidification and the benzoate transcriptome in Bacillus subtilis.

Ryan D Kitko1, Rebecca L Cleeton, Erin I Armentrout

  • 1Department of Biology, Kenyon College, Gambier, Ohio, United States of America.

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|December 17, 2009
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Summary

Bacillus subtilis maintains cytoplasmic pH homeostasis during acid stress, adapting to high benzoate concentrations better than E. coli. This study reveals key genes involved in pH response and adaptation.

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Area of Science:

  • Microbiology
  • Bacterial Physiology
  • Gene Regulation

Background:

  • Bacillus subtilis must maintain stable internal pH despite fluctuating external conditions.
  • Understanding how bacteria respond to acid stress and permeant acids is crucial.
  • Mechanisms of cytoplasmic pH regulation in bacteria are not fully understood.

Purpose of the Study:

  • To investigate the cytoplasmic pH response of Bacillus subtilis to rapid external acidification.
  • To analyze the transcriptome of B. subtilis adapted to benzoate stress.
  • To compare the acid stress tolerance of B. subtilis with E. coli.

Main Methods:

  • Real-time measurement of cytoplasmic pH using GFPmut3b fluorimetry.
  • Exposure of B. subtilis to rapid external pH changes and benzoate.
  • Transcriptome analysis using gene expression indices to identify differentially expressed genes.

Main Results:

  • B. subtilis exhibited cytoplasmic acidification and partial recovery upon external acidification.
  • Growth with 30 mM benzoate depressed cytoplasmic pH but allowed for adaptation, exceeding E. coli's tolerance.
  • 164 genes were upregulated and 102 downregulated by benzoate, with significant overlap with acid-stress response genes.

Conclusions:

  • B. subtilis demonstrates robust pH homeostasis and tolerance to permeant acid stress.
  • The benzoate adaptation transcriptome shares common elements with the acid stress transcriptome.
  • Cytoplasmic pH likely plays a role in regulating specific gene clusters, including the SigW regulon.