Cargando…

The Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells

Colibactins are hybrid polyketide-nonribosomal peptides produced by Escherichia coli, Klebsiella pneumoniae, and other Enterobacteriaceae harboring the pks genomic island. These genotoxic metabolites are produced by pks-encoded peptide-polyketide synthases as inactive prodrugs called precolibactins,...

Descripción completa

Detalles Bibliográficos
Autores principales: Bossuet-Greif, Nadège, Vignard, Julien, Taieb, Frédéric, Mirey, Gladys, Dubois, Damien, Petit, Claude, Oswald, Eric, Nougayrède, Jean-Philippe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5874909/
https://www.ncbi.nlm.nih.gov/pubmed/29559578
http://dx.doi.org/10.1128/mBio.02393-17
_version_ 1783310258665422848
author Bossuet-Greif, Nadège
Vignard, Julien
Taieb, Frédéric
Mirey, Gladys
Dubois, Damien
Petit, Claude
Oswald, Eric
Nougayrède, Jean-Philippe
author_facet Bossuet-Greif, Nadège
Vignard, Julien
Taieb, Frédéric
Mirey, Gladys
Dubois, Damien
Petit, Claude
Oswald, Eric
Nougayrède, Jean-Philippe
author_sort Bossuet-Greif, Nadège
collection PubMed
description Colibactins are hybrid polyketide-nonribosomal peptides produced by Escherichia coli, Klebsiella pneumoniae, and other Enterobacteriaceae harboring the pks genomic island. These genotoxic metabolites are produced by pks-encoded peptide-polyketide synthases as inactive prodrugs called precolibactins, which are then converted to colibactins by deacylation for DNA-damaging effects. Colibactins are bona fide virulence factors and are suspected of promoting colorectal carcinogenesis when produced by intestinal E. coli. Natural active colibactins have not been isolated, and how they induce DNA damage in the eukaryotic host cell is poorly characterized. Here, we show that DNA strands are cross-linked covalently when exposed to enterobacteria producing colibactins. DNA cross-linking is abrogated in a clbP mutant unable to deacetylate precolibactins or by adding the colibactin self-resistance protein ClbS, confirming the involvement of the mature forms of colibactins. A similar DNA-damaging mechanism is observed in cellulo, where interstrand cross-links are detected in the genomic DNA of cultured human cells exposed to colibactin-producing bacteria. The intoxicated cells exhibit replication stress, activation of ataxia-telangiectasia and Rad3-related kinase (ATR), and recruitment of the DNA cross-link repair Fanconi anemia protein D2 (FANCD2) protein. In contrast, inhibition of ATR or knockdown of FANCD2 reduces the survival of cells exposed to colibactin-producing bacteria. These findings demonstrate that DNA interstrand cross-linking is the critical mechanism of colibactin-induced DNA damage in infected cells.
format Online
Article
Text
id pubmed-5874909
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher American Society for Microbiology
record_format MEDLINE/PubMed
spelling pubmed-58749092018-03-29 The Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells Bossuet-Greif, Nadège Vignard, Julien Taieb, Frédéric Mirey, Gladys Dubois, Damien Petit, Claude Oswald, Eric Nougayrède, Jean-Philippe mBio Research Article Colibactins are hybrid polyketide-nonribosomal peptides produced by Escherichia coli, Klebsiella pneumoniae, and other Enterobacteriaceae harboring the pks genomic island. These genotoxic metabolites are produced by pks-encoded peptide-polyketide synthases as inactive prodrugs called precolibactins, which are then converted to colibactins by deacylation for DNA-damaging effects. Colibactins are bona fide virulence factors and are suspected of promoting colorectal carcinogenesis when produced by intestinal E. coli. Natural active colibactins have not been isolated, and how they induce DNA damage in the eukaryotic host cell is poorly characterized. Here, we show that DNA strands are cross-linked covalently when exposed to enterobacteria producing colibactins. DNA cross-linking is abrogated in a clbP mutant unable to deacetylate precolibactins or by adding the colibactin self-resistance protein ClbS, confirming the involvement of the mature forms of colibactins. A similar DNA-damaging mechanism is observed in cellulo, where interstrand cross-links are detected in the genomic DNA of cultured human cells exposed to colibactin-producing bacteria. The intoxicated cells exhibit replication stress, activation of ataxia-telangiectasia and Rad3-related kinase (ATR), and recruitment of the DNA cross-link repair Fanconi anemia protein D2 (FANCD2) protein. In contrast, inhibition of ATR or knockdown of FANCD2 reduces the survival of cells exposed to colibactin-producing bacteria. These findings demonstrate that DNA interstrand cross-linking is the critical mechanism of colibactin-induced DNA damage in infected cells. American Society for Microbiology 2018-03-20 /pmc/articles/PMC5874909/ /pubmed/29559578 http://dx.doi.org/10.1128/mBio.02393-17 Text en Copyright © 2018 Bossuet-Greif et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Bossuet-Greif, Nadège
Vignard, Julien
Taieb, Frédéric
Mirey, Gladys
Dubois, Damien
Petit, Claude
Oswald, Eric
Nougayrède, Jean-Philippe
The Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells
title The Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells
title_full The Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells
title_fullStr The Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells
title_full_unstemmed The Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells
title_short The Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells
title_sort colibactin genotoxin generates dna interstrand cross-links in infected cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5874909/
https://www.ncbi.nlm.nih.gov/pubmed/29559578
http://dx.doi.org/10.1128/mBio.02393-17
work_keys_str_mv AT bossuetgreifnadege thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT vignardjulien thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT taiebfrederic thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT mireygladys thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT duboisdamien thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT petitclaude thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT oswalderic thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT nougayredejeanphilippe thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT bossuetgreifnadege colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT vignardjulien colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT taiebfrederic colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT mireygladys colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT duboisdamien colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT petitclaude colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT oswalderic colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
AT nougayredejeanphilippe colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells