title: Response of Escherichia coli growth rate to osmotic shock.
abstract: It has long been proposed that turgor pressure plays an essential role during bacterial growth by driving mechanical expansion of the cell wall. This hypothesis is based on analogy to plant cells, for which this mechanism has been established, and on experiments in which the growth rate of bacterial cultures was observed to decrease as the osmolarity of the growth medium was increased. To distinguish the effect of turgor pressure from pressure-independent effects that osmolarity might have on cell growth, we monitored the elongation of single Escherichia coli cells while rapidly changing the osmolarity of their media. By plasmolyzing cells, we found that cell-wall elastic strain did not scale with growth rate, suggesting that pressure does not drive cell-wall expansion. Furthermore, in response to hyper- and hypoosmotic shock, E. coli cells resumed their preshock growth rate and relaxed to their steady-state rate after several minutes, demonstrating that osmolarity modulates growth rate slowly, independently of pressure. Oscillatory hyperosmotic shock revealed that although plasmolysis slowed cell elongation, the cells nevertheless "stored" growth such that once turgor was reestablished the cells elongated to the length that they would have attained had they never been plasmolyzed. Finally, MreB dynamics were unaffected by osmotic shock. These results reveal the simple nature of E. coli cell-wall expansion: that the rate of expansion is determined by the rate of peptidoglycan insertion and insertion is not directly dependent on turgor pressure, but that pressure does play a basic role whereby it enables full extension of recently inserted peptidoglycan.
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA 94305; andDepartment of Biochemistry,Howard Hughes Medical Institute, and.
authors.forename: Enrique
authors.initials: E
authors.lastname: Rojas
authors.affiliation: Department of Biochemistry,Howard Hughes Medical Institute, andDepartment of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305 theriot@stanford.edu kchuang@stanford.edu.
authors.forename: Julie A
authors.initials: JA
authors.lastname: Theriot
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA 94305; andDepartment of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305 theriot@stanford.edu kchuang@stanford.edu.
authors.forename: Kerwyn Casey
authors.initials: KC
authors.lastname: Huang
mesh_terms: D000220:Adaptation, Biological / Q000502:physiology*; D001696:Biomechanical Phenomena; D002473:Cell Wall / Q000502:physiology*; D004548:Elasticity; D004926:Escherichia coli / Q000254:growth & development*; D008856:Microscopy, Fluorescence; D009024:Morphogenesis / Q000502:physiology*; D009997:Osmotic Pressure / Q000502:physiology*; D011312:Pressure; D059008:Time-Lapse Imaging
keywords: bacterial morphogenesis; cell mechanics