Cargando…

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

Immune cells kill invading microbes by producing reactive oxygen and nitrogen species, primarily hydrogen peroxide (H(2)O(2)) and nitric oxide (NO). We previously found that NO inhibits catalases in Escherichia coli, stabilizing H(2)O(2) around treated cells and promoting catastrophic chromosome fra...

Descripción completa

Detalles Bibliográficos
Autores principales: Agashe, Pooja, Kuzminov, Andrei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9018393/
https://www.ncbi.nlm.nih.gov/pubmed/35288189
http://dx.doi.org/10.1016/j.jbc.2022.101825
_version_ 1784689040772038656
author Agashe, Pooja
Kuzminov, Andrei
author_facet Agashe, Pooja
Kuzminov, Andrei
author_sort Agashe, Pooja
collection PubMed
description Immune cells kill invading microbes by producing reactive oxygen and nitrogen species, primarily hydrogen peroxide (H(2)O(2)) and nitric oxide (NO). We previously found that NO inhibits catalases in Escherichia coli, stabilizing H(2)O(2) around treated cells and promoting catastrophic chromosome fragmentation via continuous Fenton reactions generating hydroxyl radicals. Indeed, H(2)O(2)-alone treatment kills catalase-deficient (katEG) mutants similar to H(2)O(2)+NO treatment. However, the Fenton reaction, in addition to H(2)O(2), requires Fe(II), which H(2)O(2) excess instantly converts into Fenton-inert Fe(III). For continuous Fenton when H(2)O(2) is stable, a supply of reduced iron becomes necessary. We show here that this supply is ensured by Fe(II) recruitment from ferritins and Fe(III) reduction by flavin reductase. Our observations also concur with NO-mediated respiration inhibition that drives Fe(III) reduction. We modeled this NO-mediated inhibition via inactivation of ndh and nuo respiratory enzymes responsible for the step of NADH oxidation, which results in increased NADH pools driving flavin reduction. We found that, like the katEG mutant, the ndh nuo double mutant is similarly sensitive to H(2)O(2)-alone and H(2)O(2)+NO treatments. Moreover, the quadruple katEG ndh nuo mutant lacking both catalases and efficient respiration was rapidly killed by H(2)O(2)-alone, but this killing was delayed by NO, rather than potentiated by it. Taken together, we conclude that NO boosts the levels of both H(2)O(2) and Fe(II) Fenton reactants, making continuous hydroxyl-radical production feasible and resulting in irreparable oxidative damage to the chromosome.
format Online
Article
Text
id pubmed-9018393
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher American Society for Biochemistry and Molecular Biology
record_format MEDLINE/PubMed
spelling pubmed-90183932022-04-22 Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli Agashe, Pooja Kuzminov, Andrei J Biol Chem Research Article Immune cells kill invading microbes by producing reactive oxygen and nitrogen species, primarily hydrogen peroxide (H(2)O(2)) and nitric oxide (NO). We previously found that NO inhibits catalases in Escherichia coli, stabilizing H(2)O(2) around treated cells and promoting catastrophic chromosome fragmentation via continuous Fenton reactions generating hydroxyl radicals. Indeed, H(2)O(2)-alone treatment kills catalase-deficient (katEG) mutants similar to H(2)O(2)+NO treatment. However, the Fenton reaction, in addition to H(2)O(2), requires Fe(II), which H(2)O(2) excess instantly converts into Fenton-inert Fe(III). For continuous Fenton when H(2)O(2) is stable, a supply of reduced iron becomes necessary. We show here that this supply is ensured by Fe(II) recruitment from ferritins and Fe(III) reduction by flavin reductase. Our observations also concur with NO-mediated respiration inhibition that drives Fe(III) reduction. We modeled this NO-mediated inhibition via inactivation of ndh and nuo respiratory enzymes responsible for the step of NADH oxidation, which results in increased NADH pools driving flavin reduction. We found that, like the katEG mutant, the ndh nuo double mutant is similarly sensitive to H(2)O(2)-alone and H(2)O(2)+NO treatments. Moreover, the quadruple katEG ndh nuo mutant lacking both catalases and efficient respiration was rapidly killed by H(2)O(2)-alone, but this killing was delayed by NO, rather than potentiated by it. Taken together, we conclude that NO boosts the levels of both H(2)O(2) and Fe(II) Fenton reactants, making continuous hydroxyl-radical production feasible and resulting in irreparable oxidative damage to the chromosome. American Society for Biochemistry and Molecular Biology 2022-03-11 /pmc/articles/PMC9018393/ /pubmed/35288189 http://dx.doi.org/10.1016/j.jbc.2022.101825 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Agashe, Pooja
Kuzminov, Andrei
Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli
title Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli
title_full Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli
title_fullStr Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli
title_full_unstemmed Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli
title_short Nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and Fe(II) Fenton reactants in E. coli
title_sort nitric oxide precipitates catastrophic chromosome fragmentation by bolstering both hydrogen peroxide and fe(ii) fenton reactants in e. coli
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9018393/
https://www.ncbi.nlm.nih.gov/pubmed/35288189
http://dx.doi.org/10.1016/j.jbc.2022.101825
work_keys_str_mv AT agashepooja nitricoxideprecipitatescatastrophicchromosomefragmentationbybolsteringbothhydrogenperoxideandfeiifentonreactantsinecoli
AT kuzminovandrei nitricoxideprecipitatescatastrophicchromosomefragmentationbybolsteringbothhydrogenperoxideandfeiifentonreactantsinecoli