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Persisters are dormant phenotypic variants of bacterial cells that are tolerant to killing by antibiotics 1 . Persisters are associated with chronic infections and antibiotic treatment failure 1-3 . In Escherichia coli , toxin/antitoxin (TA) modules have been linked to persister formation 4-6 . The mechanism of persister formation in Gram-positive bacteria is unknown. Staphylococcus aureus is a major human pathogen, responsible for a variety of chronic and relapsing infections such as osteomyelitis, endocarditis and infections of implanted devices. Deleting TA modules in S. aureus did not affect the level of persisters. Here we show that S. aureus persisters are produced due to a stochastic entrance into stationary phase accompanied by a drop in intracellular ATP. Cells expressing stationary state markers are present throughout the growth phase, increasing in frequency with cell density. Cell sorting revealed that expression of stationary markers is associated with a 100-1000 fold increase in the likelihood of survival to antibiotic challenge. The ATP level of the cell is predictive of bactericidal antibiotic efficacy and explains bacterial tolerance to antibiotics.

Persister formation in Staphylococcus aureus is associated with ATP depletion