Proteomics Study of Filamentous Persisters from Evo3A Evolved Populations during Ampicillin Treatment and Resuscitation

Proteomics is the most suitable tool to study persisters with their complex underlying molecular mechanisms from a system-level perspective, but the number of persisters that present naturally is too few for proteomics analysis. Here, we utilized Evo3A, an evolved population with enriched persisters fraction from a recent adaptive laboratory evolution experiment, to study the mechanisms of persistence during ampicillin treatment and resuscitation. Interestingly, the enriched persisters on Evo3A exhibit filamentous morphology upon treatment with ampicillin, and the filaments are getting longer over time. Time-course proteomics study revealed that proteins involved in major carbohydrate metabolism are up-regulated, in particular those involved in the oxidative stress response and act as cellular response to DNA damage. As opposed to the proteome profile during antibiotic treatment, proteins involved in major metabolic processes and ATP generation are down-regulated, while translational proteins and porins are up-regulated in the filaments during resuscitation.