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Cell-shape homeostasis in Escherichia coli is driven by growth, division, and nucleoid complexity.

ABSTRACT: Analysis of recently published high-throughput measurements of wild-type Escherichia coli cells growing at a wide range of rates demonstrates that cell width W, which is constant at any particular growth rate, is related (with a CV = 2.4%) to the level of nucleoid complexity, expressed as the amount of DNA in genome equivalents that is associated with chromosome terminus (G/terC). The relatively constant (CV = 7.3%) aspect ratio of newborn cells (Lb/W) in populations growing at different rates indicates existence of cell-shape homeostasis. Enlarged W of thymine-limited thyA mutants growing at identical rates support the hypothesis that nucleoid complexity actively affects W. Nucleoid dynamics is proposed to transmit a primary signal to the peptidoglycan-synthesizing system through the transertion mechanism, i.e., coupled transcription/translation of genes encoding membrane proteins and inserting these proteins into the membrane.

SUBMITTER: Zaritsky A

PROVIDER: S-EPMC4623199 | biostudies-literature | 2015 Jul

REPOSITORIES: biostudies-literature

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Cell-shape homeostasis in Escherichia coli is driven by growth, division, and nucleoid complexity.

Zaritsky Arieh A

Biophysical journal 20150701 2

Analysis of recently published high-throughput measurements of wild-type Escherichia coli cells growing at a wide range of rates demonstrates that cell width W, which is constant at any particular growth rate, is related (with a CV = 2.4%) to the level of nucleoid complexity, expressed as the amount of DNA in genome equivalents that is associated with chromosome terminus (G/terC). The relatively constant (CV = 7.3%) aspect ratio of newborn cells (Lb/W) in populations growing at different rates i ...[more]

PMID: 26200854

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Project description:BackgroundA fundamental characteristic of cells is the ability to divide. To date, most parameters of bacterial cultures, including cell division, have been measured as cell population averages, assuming that all bacteria divide at a uniform rate.ResultsWe monitored the division of individual cells in Escherichia coli cultures during different growth phases. Our experiments are based on the dilution of green fluorescent protein (GFP) upon cell division, monitored by flow cytometry. The results show that the vast majority of E. coli cells in exponentially growing cultures divided uniformly. In cultures that had been in stationary phase up to four days, no cell division was observed. However, upon dilution of stationary phase culture into fresh medium, two subpopulations of cells emerged: one that started dividing and another that did not. These populations were detectable by GFP dilution and displayed different side scatter parameters in flow cytometry. Further analysis showed that bacteria in the non-growing subpopulation were not dead, neither was the difference in growth capacity reducible to differences in stationary phase-specific gene expression since we observed uniform expression of several stress-related promoters. The presence of non-growing persisters, temporarily dormant bacteria that are tolerant to antibiotics, has previously been described within growing bacterial populations. Using the GFP dilution method combined with cell sorting, we showed that ampicillin lyses growing bacteria while non-growing bacteria retain viability and that some of them restart growth after the ampicillin is removed. Thus, our method enables persisters to be monitored even in liquid cultures of wild type strains in which persister formation has low frequency.ConclusionIn principle, the approaches developed here could be used to detect differences in cell division in response to different environmental conditions and in cultures of unicellular organisms other than E. coli.

Dataset Information

Cell-shape homeostasis in Escherichia coli is driven by growth, division, and nucleoid complexity.

Publications

Cell-shape homeostasis in Escherichia coli is driven by growth, division, and nucleoid complexity.

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