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A fraction of otherwise antimicrobial-sensitive  Bacillus subtilis  cells, called persisters, are phenotypically tolerant of antimicrobial treatment. We report that, independently of  B. subtilis'  growth phase, transient ζ toxin expression induces a dormant state and alters cellular responses so that cells are more sensitive to antimicrobials with different modes of action. This outcome is modulated by fine tuning (p)ppGpp and GTP levels: i) in the presence of low “dysregulated” (p)ppGpp levels (as in  relA   −  cells) hyper-tolerance to both toxin and antimicrobials was observed; ii) physiological or low (p)ppGpp levels (as in the wild-type,  sasA   − ,  sasB   −  and  relA   −   sasA   −  context) show a normal toxin and antimicrobial tolerance; and iii) lower levels (in  relA   −   sasB   − ) or absence of (p)ppGpp (in the  relA   −   sasA   −   sasB   −  context), in concert with elevated GTP levels, potentiate the efficacy of both toxin and antimicrobial action, rendering tolerance vulnerable to eradication.

Role of Toxin ζ and Starvation Responses in the Sensitivity to Antimicrobials