@article{8659641,
  abstract     = {{Antibiotic tolerance characterized by slow killing of bacteria in response to a drug can lead to treatment failure and promote the emergence of resistance. beta-lactam antibiotics inhibit cell wall growth in bacteria and many of them cause filamentation followed by cell lysis. Hence delayed cell lysis can lead to beta-lactam tolerance. Systematic discovery of genetic factors that affect beta-lactam killing kinetics has not been performed before due to challenges in high-throughput, dynamic analysis of viability of filamented cells during bactericidal action. We implemented a high-throughput time-resolved microscopy approach in a gene deletion library of Escherichia coli to monitor the response of mutants to the beta-lactam cephalexin. Changes in frequency of lysed and intact cells due to the antibiotic action uncovered several strains with atypical lysis kinetics. Filamentation confers tolerance because antibiotic removal before lysis leads to recovery through numerous concurrent divisions of filamented cells. Filamentation-mediated tolerance was not associated with resistance, and therefore this phenotype is not discernible through most antibiotic susceptibility methods. We find that deletion of Tol-Pal proteins TolQ, TolR, or Pal but not TolA, TolB, or CpoB leads to rapid killing by beta-lactams. We also show that the timing of cell wall degradation determines the lysis and killing kinetics after beta-lactam treatment. Altogether, this study uncovers numerous genetic determinants of hitherto unappreciated filamentation-mediated beta-lactam tolerance and support the growing call for considering antibiotic tolerance in clinical evaluation of pathogens. More generally, the microscopy screening methodology described here can easily be adapted to study lysis in large numbers of strains.}},
  articleno    = {{374}},
  author       = {{Zahir, Taiyeb and Wilmaerts, Dorien and Franke, Sabine and Weytjens, Bram and Camacho, Rafael and Marchal, Kathleen and Hofkens, Johan and Fauvart, Maarten and Michielsen, Jan}},
  issn         = {{1664-302X}},
  journal      = {{FRONTIERS IN MICROBIOLOGY}},
  keywords     = {{PENICILLIN-BINDING-PROTEINS,MULTIPLE-ANTIBIOTIC-RESISTANCE,OUTER-MEMBRANE PERMEABILITY,ESCHERICHIA-COLI,PSEUDOMONAS-AERUGINOSA,BACTERIAL SURVIVAL,CELL-SEPARATION,MECHANISM,DIVISION,MUTANTS,beta-lactam,antibiotic tolerance,filamentation,high-throughput,microscopy,bacteriolysis,β-lactam}},
  language     = {{eng}},
  pages        = {{14}},
  title        = {{Image-based dynamic phenotyping reveals genetic determinants of filamentation-mediated beta-lactam tolerance}},
  url          = {{http://doi.org/10.3389/fmicb.2020.00374}},
  volume       = {{11}},
  year         = {{2020}},
}

Altmetric

View in Altmetric

Web of Science

Times cited: