Contribution of SOS genes to H2O2-induced apoptosis-like death in Escherichia coli.

Abstract

Hydrogen peroxide (H₂O₂) is a debriding agent that damages the microbial structure and function by generating various reactive oxygen species (ROS). H₂O₂-produced hydroxyl radical (OH∙) also exerts oxidative stress on microorganisms. The spread of antibiotic-resistance in bacteria is a serious issue worldwide, and greater efforts are needed to identify and characterize novel antibacterial mechanisms to develop new treatment strategies. Therefore, this study aimed to clarify the relationship between H₂O₂ and Escherichia coli and to elucidate a novel antibacterial mechanism(s) of H₂O₂. Following H₂O₂ exposure, increased levels of 8-hydroxydeoxyguanosine and malondialdehyde indicated that H₂O₂ accelerates oxidation of bacterial DNA and lipids in E. coli. As oxidative damage worsened, the SOS response was triggered. Cell division arrest and resulting filamentous cells were identified in cells, indicating that LexA was involved in DNA replication. It was also verified that RecA, a representative SOS gene, helps self-cleavage of LexA and acts as a bacterial caspase-like protein. Our findings also showed that dinF is essential to preserve E. coli from H₂O₂-induced ROS, and furthermore, demonstrated that H₂O₂-induced SOS response and SOS genes participate differently in guarding E. coli from oxidative stress. As an extreme SOS response is considered apoptosis-like death (ALD) in bacteria, additional experiments were performed to examine the characteristics of ALD. DNA fragmentation and membrane depolarization appeared in H₂O₂-treated cells, suggesting that H₂O₂ causes ALD in E. coli. In conclusion, our investigations revealed that ALD is a novel antibacterial mode of action(s) of H₂O₂ with important contributions from SOS genes.