title: Metabolic control of persister formation in Escherichia coli.
abstract: Bacterial persisters are phenotypic variants that form from the action of stress response pathways triggering toxin-mediated antibiotic tolerance. Although persisters form during normal growth from native stresses, the pathways responsible for this phenomenon remain elusive. Here we have discovered that carbon source transitions stimulate the formation of fluoroquinolone persisters in Escherichia coli. Further, through a combination of genetic, biochemical, and flow cytometric assays in conjunction with a mathematical model, we have reconstructed a molecular-level persister formation pathway from initial stress (glucose exhaustion) to the activation of a metabolic toxin-antitoxin (TA) module (the ppGpp biochemical network) resulting in inhibition of DNA gyrase activity, the primary target of fluoroquinolones. This pathway spans from initial stress to antibiotic target and demonstrates that TA behavior can be exhibited by a metabolite-enzyme interaction (ppGpp-SpoT), in contrast to classical TA systems that involve only protein and/or RNA.
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA.
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Amato
authors.forename: Mehmet A
authors.initials: MA
authors.lastname: Orman
authors.forename: Mark P
authors.initials: MP
authors.lastname: Brynildsen
mesh_terms: D000222:Adaptation, Physiological / Q000187:drug effects* / Q000235:genetics; D000667:Ampicillin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D000992:Antitoxins / Q000235:genetics / Q000378:metabolism; D058491:Bacterial Load; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism; D002244:Carbon / Q000378:metabolism*; D000242:Cyclic AMP / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D024841:Fluoroquinolones / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D006159:Guanosine Tetraphosphate / Q000378:metabolism; D050296:Microbial Viability / Q000187:drug effects / Q000235:genetics; D015242:Ofloxacin / Q000494:pharmacology; D011755:Pyrophosphatases / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents; D000992:Antitoxins; D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D024841:Fluoroquinolones; D006159:Guanosine Tetraphosphate; D002244:Carbon; D000667:Ampicillin; D015242:Ofloxacin; D000242:Cyclic AMP; C027627:guanosine-3',5'-bis(diphosphate) 3'-pyrophosphatase; D011755:Pyrophosphatases; D005947:Glucose