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De novo morphogenesis in L‐forms via geometric control of cell growth
Authors:Gabriel Billings  Nikolay Ouzounov  Tristan Ursell  Samantha M Desmarais  Joshua Shaevitz  Zemer Gitai  Kerwyn Casey Huang
Institution:1. Department of Physics, Stanford University, , Stanford, CA, 94305 USA;2. Department of Molecular Biology, Princeton University, , Princeton, NJ, 08544 USA;3. Department of Bioengineering, Stanford University, , Stanford, CA, 94305 USA;4. Department of Physics, Princeton University, , Princeton, NJ, 08544 USA;5. Lewis‐Sigler Institute for Integrative Genomics, Princeton University, , Princeton, NJ, 08544 USA;6. Department of Microbiology and Immunology, Stanford University School of Medicine, , Stanford, CA, 94305 USA
Abstract:In virtually all bacteria, the cell wall is crucial for mechanical integrity and for determining cell shape. Escherichia coli's rod‐like shape is maintained via the spatiotemporal patterning of cell‐wall synthesis by the actin homologue MreB. Here, we transiently inhibited cell‐wall synthesis in E. coli to generate cell‐wall‐deficient, spherical L‐forms, and found that they robustly reverted to a rod‐like shape within several generations after inhibition cessation. The chemical composition of the cell wall remained essentially unchanged during this process, as indicated by liquid chromatography. Throughout reversion, MreB localized to inwardly curved regions of the cell, and fluorescent cell wall labelling revealed that MreB targets synthesis to those regions. When exposed to the MreB inhibitor A22, reverting cells regrew a cell wall but failed to recover a rod‐like shape. Our results suggest that MreB provides the geometric measure that allows E. coli to actively establish and regulate its morphology.
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