title: Distinct single-cell morphological dynamics under beta-lactam antibiotics.
abstract: The bacterial cell wall is conserved in prokaryotes, stabilizing cells against osmotic stress. Beta-lactams inhibit cell-wall synthesis and induce lysis through a bulge-mediated mechanism; however, little is known about the formation dynamics and stability of these bulges. To capture processes of different timescales, we developed an imaging platform combining automated image analysis with live-cell microscopy at high time resolution. Beta-lactam killing of Escherichia coli cells proceeded through four stages: elongation, bulge formation, bulge stagnation, and lysis. Both the cell wall and outer membrane (OM) affect the observed dynamics; damaging the cell wall with different beta-lactams and compromising OM integrity cause different modes and rates of lysis. Our results show that the bulge-formation dynamics are determined by how the cell wall is perturbed. The OM plays an independent role in stabilizing the bulge once it is formed. The stabilized bulge delays lysis and allows recovery upon drug removal.
authors.affiliation: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
authors.forename: Zhizhong
authors.initials: Z
authors.lastname: Yao
authors.forename: Daniel
authors.initials: D
authors.lastname: Kahne
authors.forename: Roy
authors.initials: R
authors.lastname: Kishony
mesh_terms: D000667:Ampicillin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D057205:Automation, Laboratory; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D002441:Cefsulodin / Q000494:pharmacology; D002473:Cell Wall / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D002506:Cephalexin / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D009154:Mutation; D013997:Time Factors; D059008:Time-Lapse Imaging; D047090:beta-Lactams / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D047090:beta-Lactams; D000667:Ampicillin; D002506:Cephalexin; D002441:Cefsulodin