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Current Genetics

, Volume 65, Issue 1, pp 167–177 | Cite as

Arenicin-1-induced apoptosis-like response requires RecA activation and hydrogen peroxide against Escherichia coli

  • Heejeong Lee
  • Dong Gun LeeEmail author
Original Article

Abstract

Arenicin-1, a 21-mer antimicrobial peptide exerts significant broad-spectrum antimicrobial activity with membrane-active mechanisms. However, owing to multiple mechanisms of cell death, the antibacterial mechanism of arenicin-1 requires detailed analysis. In the present study, arenicin-1-treated bacteria underwent an apoptosis-like response, which was mechanistically and morphologically similar to eukaryotic apoptosis. Changes in the physiological status of arenicin-1-treated bacterial cells involved accumulation of reactive oxygen species, imbalance of intracellular calcium gradients, disruption of membrane potential, bacterial caspase-like protein activation, and DNA damage. In arenicin-1-induced apoptosis-like death, autocleavage of LexA was observed because of the activation of the caspase-like activity of RecA. Additionally, typical reactive oxygen species such as superoxide, hydrogen peroxide, and hydroxyl radicals, were scavenged in arenicin-1-treated cells to assess the role of specific reactive oxygen species. Scavenging of hydrogen peroxide interfered with the activity of arenicin-1 in Escherichia coli, whereas the superoxide and hydroxyl radicals level did not affect arenicin-1-induced apoptosis-like death activity. Furthermore, inhibition of Fenton reaction attenuated apoptosis-like response. In conclusion, arenicin-1-induced apoptosis like death requires SOS response proteins and is mediated by hydrogen peroxide and Fenton reaction in E. coli. Arenicin-1 may be a representative antimicrobial peptide with potent apoptotic response against E. coli.

Keywords

Arenicin-1 Hydrogen peroxide Apoptosis-like death SOS response 

Abbreviations

PCD

Programmed cell death

TLD

Thymine-less death

ALD

Apoptotic-like death

AMP

Antimicrobial peptides

PBS

Phosphate-buffered saline

GSH

Reduced glutathione

DiBAC4(3)

Bis-(1,3-dibutylbarbituric acid) trimethine oxonol

H2DCFDA

2′,7′-Dichlorodihydrofluorescein diacetate

Notes

Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korean government (MSIP) (No. 2017R1A2B4005811).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Supplementary material

294_2018_855_MOESM1_ESM.tif (9.8 mb)
Fig. S1. Oxidative stress in ΔRecA and K996 cells treated with arenicin-1. ΔRecA cells were treated with arenicin-1 (12.5 µM) or norfloxacin (2.3 µM). K996 cells were treated with arenicin-1 (12.5 µM) or norfloxacin (2.3 µM). (A) ROS production was detected via H2DCFDA staining (B) Decrease in GSH levels indicate oxidative damage. (C) Increase in MDA levels indicate lipid peroxidation. The data represents the average, standard deviation, and p values from three independent experiments (*p < 0.1; **p < 0.05; ***p < 0.01 vs the untreated; Student’s t test) (TIF 10063 KB)
294_2018_855_MOESM2_ESM.tif (11.7 mb)
Fig. S2. Features of apoptotic-like death in ΔRecA and K996 cells induced by arenicin-1. Analysis of membrane depolarization in ΔRecA and K996 cells in the presence/absence of arenicin-1 and ROS scavenger. Flow cytometric analysis of membrane depolarization using DiBAC4(3) (TIF 12006 KB)
294_2018_855_MOESM3_ESM.tif (13.6 mb)
Fig. S3 Features of apoptotic-like death in ΔRecA and K996 cells induced by arenicin-1. Analysis of cell filamentation in ΔRecA and K996 cells treated with/without arenicin-1 and ROS scavenger. Induction-mediated cell size changes. An increase in FS and SS indicated cell filamentation (TIF 13952 KB)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.School of Life Sciences, BK 21 Plus KNU Creative BioResearch Group, College of Natural SciencesKyungpook National UniversityDaeguRepublic of Korea

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