Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Regulation of cysteine homeostasis and its effect on Escherichia coli sensitivity to ciprofloxacin in LB medium

Galina Smirnova
* ORCID logo ,
Aleksey Tyulenev
,
Lyubov Sutormina
,
Tatyana Kalashnikova
ORCID logo ,
Nadezda Muzyka
,
Vadim Ushakov
,
Zoya Samoilova
,
Oleg Oktyabrsky
Version 1 : Received: 29 March 2024 / Approved: 29 March 2024 / Online: 1 April 2024 (10:18:59 CEST)

A peer-reviewed article of this Preprint also exists.

Smirnova, G.; Tyulenev, A.; Sutormina, L.; Kalashnikova, T.; Muzyka, N.; Ushakov, V.; Samoilova, Z.; Oktyabrsky, O. Regulation of Cysteine Homeostasis and Its Effect on Escherichia coli Sensitivity to Ciprofloxacin in LB Medium. Int. J. Mol. Sci. 2024, 25, 4424. Smirnova, G.; Tyulenev, A.; Sutormina, L.; Kalashnikova, T.; Muzyka, N.; Ushakov, V.; Samoilova, Z.; Oktyabrsky, O. Regulation of Cysteine Homeostasis and Its Effect on Escherichia coli Sensitivity to Ciprofloxacin in LB Medium. Int. J. Mol. Sci. 2024, 25, 4424.

Abstract

Cysteine and its derivatives, including H2S, can influence bacterial virulence and sensitivity to antibiotics. In minimal sulfate media, H2S is generated under stress to prevent excess cysteine and, together with incorporation into glutathione and export into the medium, is a mechanism of cysteine homeostasis. Here, we studied the features of cysteine homeostasis in LB medium, where the main source of sulfur is cystine, whose import can create excess cysteine inside cells. We used mutants in the mechanisms of cysteine homeostasis and a set of microbiological and biochemical methods, including real-time monitoring of sulfide and oxygen, determination of cysteine and glutathione, and expression of the Fur, OxyR, and SOS regulons genes. During normal growth, the parental strain generated H2S when switching respiration to another substrate. The mutations affected the onset time, intensity and duration of H2S production, cysteine and glutathione levels, bacterial growth and respiration rates, and induction of defense systems. Exposure to chloramphenicol and high doses of ciprofloxacin increased cysteine content and GSH synthesis. A high inverse relationship between log CFU/ml and bacterial growth rate before ciprofloxacin addition was revealed. The study points to the important role of maintaining cysteine homeostasis during normal growth and antibiotic exposure in LB medium.

Keywords

cysteine homeostasis; glutathione; H2S; Fur and SOS regulons; growth; respiration; antibiotics; bacterial survival

Subject

Biology and Life Sciences, Immunology and Microbiology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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