Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli

Aims This study aimed to establish the mechanisms of action (MOA) of a novel surface functionalised polyacrylonitrile (PAN) catalyst, which was previously shown to have potent antimicrobial activity in conjunction with hydrogen peroxide (H2O2). Methods and results Bactericidal activity was determined using a disinfectant suspension test. The MOA was investigated by measuring loss of 260 nm absorbing material, membrane potential and permeability assays, analysis of intra- and extracellular ATP and pH, and tolerance to sodium chloride and bile salts. The catalyst lowered sub-lethal concentrations of H2O2 from 0.2% to 0.09%. H2O2 ± 3 g PAN catalyst significantly (P ≤ 0.05) reduced sodium chloride and bile salt tolerance, suggesting the occurance of sublethal cell membrane damage. The catalyst significantly increased (P ≤ 0.05) N-Phenyl-l-napthylamine uptake (1.51-fold) and leakage of nucleic acids, demonstrating increased membrane permeability. A significant (P ≤ 0.05) loss of membrane potential (0.015 a.u.), coupled with pertubation of intracellular pH homeostasis and depletion of intracellular ATP, suggests potentiation of H2O2-mediated cell membrane damage. Conclusions This is the first study to investigate the catalyst's antimicrobial mechanism of action, with the cytoplasmic membrane being a target for cellular injury.

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Heterogeneous Fenton’s-like catalyst potentiation of hydrogen peroxide disinfection: an investigation into mechanisms of action

Sewell

Owen

Laird

et al. 2023

Journal of Applied Microbiology

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Bacterial nucleoid is a riddle wrapped in a mystery inside an enigma

Kuzminov

2024

J Bacteriol

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Bacterial chromosome, the nucleoid, is traditionally modeled as a rosette of DNA mega-loops, organized around proteinaceous central scaffold by nucleoid-associated proteins (NAPs), and mixed with the cytoplasm by transcription and translation. Electron microscopy of fixed cells confirms dispersal of the cloud-like nucleoid within the ribosome-filled cytoplasm. Here, I discuss evidence that the nucleoid in live cells forms DNA phase separate from riboprotein phase, the “riboid.” I argue that the nucleoid-riboid interphase, where DNA interacts with NAPs, transcribing RNA polymerases, nascent transcripts, and ssRNA chaperones, forms the transcription zone. An active part of phase separation, transcription zone enforces segregation of the centrally positioned information phase (the nucleoid) from the surrounding action phase (the riboid), where translation happens, protein accumulates, and metabolism occurs. I speculate that HU NAP mostly tiles up the nucleoid periphery—facilitating DNA mobility but also supporting transcription in the interphase. Besides extruding plectonemically supercoiled DNA mega-loops, condensins could compact them into solenoids of uniform rings, while HU could support rigidity and rotation of these DNA rings. The two-phase cytoplasm arrangement allows the bacterial cell to organize the central dogma activities, where (from the cell center to its periphery) DNA replicates and segregates, DNA is transcribed, nascent mRNA is handed over to ribosomes, mRNA is translated into proteins, and finally, the used mRNA is recycled into nucleotides at the inner membrane. The resulting information-action conveyor, with one activity naturally leading to the next one, explains the efficiency of prokaryotic cell design—even though its main intracellular transportation mode is free diffusion.

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Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli

Cited by 2 publications

References 89 publications

Heterogeneous Fenton’s-like catalyst potentiation of hydrogen peroxide disinfection: an investigation into mechanisms of action

Heterogeneous Fenton’s-like catalyst potentiation of hydrogen peroxide disinfection: an investigation into mechanisms of action

Bacterial nucleoid is a riddle wrapped in a mystery inside an enigma

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