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The sensitivity of Escherichia coli K12 to weak acid stress depends on the disruption of intramembrane anion permeability. The effects of low pH and of anion transport inhibitors were examined in three isogenic strains with alteration in the membrane anion permeability: the gap 1 mutant (sac, rfa) requires for growth a gradient of at least 0.2 pH units for cytoplasmic acidification, the phoA mutant requires an acidic environment for all cell processes, and the nut locus affects transmembrane anion transport. Mutations in the gap 1 and phoA loci make bacteria sensitive to a pH rise from 5 to 6.5, while the nut mutant is sensitive to a pH rise from 5 to 4.6. However, the growth of all strains was inhibited by a short pH shift to the growth limiting pH, with no effect of added lactic or/and acetic acid. While N-ethylmaleimide (NEM) inhibited growth at pH less than 4.8 in both phoA and phoA NEM-sensitive strains, its activity was greater in the phoA cells. The acid-resistance phenotype of the phoA mutant was suppressed by the presence of the gap 1 mutation and by pre-incubation with 4-chloromercuribenzoic acid (4-CMA) or cycloheximide at pH less than 4.8, while no restoration of the acid-sensitivity of the gap 1 mutant was observed. H+, Na+-countertransport was stimulated in phoA by cytoplasmic acidification in a nut or phoA mutant, while in the phoA NEM-sensitive mutant, there was only a slight stimulation in the
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