4.5 Article

Min Oscillations as Real-time Reporter of Sublethal Effects in Photodynamic Treatment of Bacteria

Journal

ACS INFECTIOUS DISEASES
Volume 8, Issue 1, Pages 86-90

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acsinfecdis.1c00583

Keywords

bacteria; fluorescent reporters; Min proteins; antimicrobial photodynamic therapy; real-time imaging; single-cell fluorescence microscopy

Funding

  1. Spanish Ministerio de Ciencia, Innovacion y Universidades [PGC2018-094802-B-I00, PRE2018-084983, SEV-2016-0686]
  2. Comunidad de Madrid [S2017/BMD-3867]

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This study investigates the effect of photodynamic treatment on the oscillations of MinD, a cell division regulator in Escherichia coli, using real-time single-cell fluorescence imaging. The results show that the concentration of methylene blue, a photosensitizer, disrupts the oscillation pattern of MinD. In contrast to antibiotics, photodynamic treatment abruptly interrupts the oscillation, reflecting different physiological mechanisms leading to bacterial death.
The Min protein system is a cell division regulator in Escherichia coli. Under normal growth conditions, MinD is associated with the membrane and undergoes pole-to-pole oscillations. The period of these oscillations has been previously proposed as a reporter for the bacterial physiological state at the single-cell level and has been used to monitor the response to sublethal challenges from antibiotics, temperature, or mechanical fatigue. Using real-time single-cell fluorescence imaging, we explore here the effect of photodynamic treatment on MinD oscillations. Irradiation of bacteria in the presence of the photosensitizer methylene blue disrupts the MinD oscillation pattern depending on its concentration. methylene blue disrupts the MinD oscillation pattern depending on its concentration. In contrast to antibiotics, which slow down the oscillation, photodynamic treatment results in an abrupt interruption, reflecting divergent physiological mechanisms leading to bacterial death. We show that MinD oscillations are sensitive to mild photodynamic effects that are overlooked by traditional methods, expanding the toolbox for mechanistic studies in antimicrobial photodynamic therapy.

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