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Escherichia coli cells are primed for survival before lethal antibiotic stress

Tahmina Hossain, Abhyudai Singh, View ORCID ProfileNicholas C. Butzin
doi: https://doi.org/10.1101/2022.11.11.516214
Tahmina Hossain
1Department of Biology and Microbiology; South Dakota State University; Brookings, SD, 57006; USA
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Abhyudai Singh
2Electrical & Computer Engineering; University of Delaware; Newark, DE 19716; USA
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Nicholas C. Butzin
1Department of Biology and Microbiology; South Dakota State University; Brookings, SD, 57006; USA
3Department of Chemistry and Biochemistry; South Dakota State University; Brookings, SD, 57006; USA
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  • ORCID record for Nicholas C. Butzin
  • For correspondence: nicholas.butzin@gmail.com
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Abstract

Non-genetic factors can cause significant fluctuations in gene expression levels. Regardless of growing in a stable environment, this fluctuation leads to cell-to-cell variability in an isogenic population. This phenotypic heterogeneity allows a tiny subset of bacterial cells in a population, referred to as persister cells, to tolerate long-term lethal antibiotic effects by entering into a non-dividing, metabolically altered state. One fundamental question is whether this heterogeneous persister population is due to a pre-existing genetic mutation or a result of a transiently-primed reversible cell state. To explore this, we tested clonal populations starting from a single cell using a modified Luria–Delbrück fluctuation test. Through we kept the conditions the same, the diversity in persistence level among clones was relatively consistent: varying from ∼60-100 and ∼40-70 fold for ampicillin (Amp) and apramycin (Apr), respectively. Then we divided and diluted each clone to observe whether the same clone had comparable persister levels for more than one generation. Replicates had similar persister levels even when clones were divided, diluted by 1:20, and allowed to grow for ∼5 generations. This result explicitly shows a cellular memory passed on for generations and eventually lost when cells are diluted to 1:100 and regrown (>7 generations). Our result demonstrates 1) the existence of a small population prepared for stress (“primed cells”) resulting in higher persister numbers, 2) the primed memory state is reproducible and transient, passed down for generations but eventually lost, and 3) a heterogeneous persister population is a result of a transiently-primed reversible cell state and not due to a pre-existing genetic mutation.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • A few figures were called out wrong in the text. This was fixed.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted November 16, 2022.
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Escherichia coli cells are primed for survival before lethal antibiotic stress
Tahmina Hossain, Abhyudai Singh, Nicholas C. Butzin
bioRxiv 2022.11.11.516214; doi: https://doi.org/10.1101/2022.11.11.516214
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Escherichia coli cells are primed for survival before lethal antibiotic stress
Tahmina Hossain, Abhyudai Singh, Nicholas C. Butzin
bioRxiv 2022.11.11.516214; doi: https://doi.org/10.1101/2022.11.11.516214

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