The structure of DNA in anabiotic and mummified Escherichia coli cells

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Abstract

The structural organization of DNA in “stressed” (with increased stress resistance), anabiotic and mummified cells obtained by introducing 4-hexylresorcinol in different concentrations at different stages of cell culture growth was studied using the synchrotron radiation diffraction technique. Experimental studies allow us to conclude that 4-hexylresorcinol is the initiator of the transition of cells into an anabiotic and mummified state in the stationary stage of growth. In the prestationary stage, in the studied concentration range, 4-hexylresorcinol initiates the transition of cells into a mummified state, but not into an anabiotic state, which indicates that DNA is unprepared for the crystallization process in these bacteria. The structure of DNA inside a cell in an anabiotic dormant state (almost complete absence of metabolism) and dormant state (starvation stress) coincide (form nanocrystalline structures). Data indicate the universality of DNA condensation or the universality of DNA protection by the Dps protein in the dormant state, regardless of the type of stress. The mummified state (complete absence of metabolism, irreversible to life) is very different in structure (has no order within the cell).

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About the authors

Yu. F. Krupyanskii

Semenov Federal Research Center of Chemical Physics, Russian Academy of Sciences

Author for correspondence.
Email: yufk@chph.ras.ru
Russian Federation, Moscow

V. V. Kovalenko

Semenov Federal Research Center of Chemical Physics, Russian Academy of Sciences

Email: yufk@chph.ras.ru
Russian Federation, Moscow

N. G. Loiko

Semenov Federal Research Center of Chemical Physics, Russian Academy of Sciences; Federal Research Center “Fundamentals of Biotechnology”, Russian Academy of Sciences

Email: yufk@chph.ras.ru
Russian Federation, Moscow; Moscow

E. V. Tereshkin

Semenov Federal Research Center of Chemical Physics, Russian Academy of Sciences

Email: yufk@chph.ras.ru
Russian Federation, Moscow

K. B. Tereshkina

Semenov Federal Research Center of Chemical Physics, Russian Academy of Sciences

Email: yufk@chph.ras.ru
Russian Federation, Moscow

A. N. Popov

European Synchrotron Radiation Facility

Email: yufk@chph.ras.ru
France, Grenoble Cedex 9

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2. Fig. 1. Scattering intensity as a function of the 2θ angle for samples containing bacterial cells of the E. coli Gold strain exposed to 4HR at the stationary growth phase at the following concentrations: a – 10–4 M, anabiotic state of the cell (curve 1, blue), 10–3 M – mummified state (curve 2, red); b – 10–4 M (curve 1, blue), 10–5 M (curve 2, purple), 10–6 M (curve 3, green). Thin yellow curves (in this figure and below) are the scattering intensity from cells in the active growth phase. The insets show the diffraction patterns of the anabiotic and mummified states of the cells.

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3. Fig. 2. Dependence of the scattering intensity on the 2θ angle of samples containing bacterial cells of the E. coli Gold strain exposed to 4HR at the pre-stationary growth phase at the following concentrations: a – 10–4 M (curve 1, red), 10–3 M (curve 2, blue); b – 10–4 M (curve 1, red), 10–5 M (curve 2, purple), 10–6 M (curve 3, green).

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4. Fig. 3. Dependence of the scattering intensity on the 2θ angle of samples containing bacterial cells of the E. coli Gold strain, exposed to 4HR at a concentration of 10–3 M at the stationary growth phase and incubated for 1.5 h (curve 1, blue), 9 h (curve 2, purple) and 100 h (curve 3, green).

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5. Fig. 4. Dynamics of germination of anabiotic E. coli cells obtained under the action of 4HR at a concentration of 10–4 M in the stationary growth phase. Incubation period of cells after transfer to a new nutrient medium: a – 0 h, b – 1.5 h, c – 14 h, d – 96 h. Curves 1 – dependence of scattering intensity on the 2θ angle of samples containing anabiotic bacterial cells; curves 2 – scattering intensity from cells in the active growth phase.

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