Proteome dynamics during antibiotic persistence and resuscitation

Abstract

Bacterial persister cells become transiently tolerant to antibiotics by restraining their growth and metabolic activity. Detailed molecular characterization of bacterial persistence is hindered by low count of persisting cells and the need for their isolation. Here we used sustained addition of stable isotope-labeled lysine to selectively label the proteome of hipA-induced persisting and hipB-induced resuscitating E. coli cells in minimal medium after antibiotic treatment. Timeresolved, 24-hour measurement of label incorporation allowed detection of over 500 newly synthetized proteins in persister cells, demonstrating low but widespread protein synthesis. Many essential proteins were newly synthesized and several ribosome-associated proteins showed unusually high synthesis levels, pointing to their roles in maintenance of persistence. At the onset of resuscitation, cells synthesized ABC transporters, restored translation machinery and resumed metabolism by inducing glycolysis and biosynthesis of amino acids. This dataset provides an unprecedented insight into the processes governing persistence and resuscitation of bacterial cells.

Proteomics in antibiotic resistance and tolerance research: Mapping the resistome and the tolerome of bacterial pathogens

“…Among these, the nuo operon has been identified as a genetic determinant of persistence and was observed to be mutated in multiple tolerant strains [147,[155][156][157]. Besides, RaiA was also found to be synthesized at a high level on E. coli persisters by Semanjski et al [141], and played an important role in forming persisters by Wood and Song [158].…”

“…A more recent study by Semanjski et al. [141] used stable isotope‐labeled lysine to label the E. coli proteome during hipA ‐induced persistence and hipB ‐induced resuscitation. Time‐resolved, 24 h measurement of label incorporation detected around 500 newly synthesized proteins in the persister cells during antibiotic treatment and revealed that several ribosome‐associated proteins such as RaiA and Sra showed high synthesis levels, highlighting their roles in persistence maintenance.…”

“…Besides, RaiA was also found to be synthesized at a high level on E. coli persisters by Semanjski et al. [141], and played an important role in forming persisters by Wood and Song [158].…”