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

ABSTRACT 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 precol...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Nadège Bossuet-Greif, Julien Vignard, Frédéric Taieb, Gladys Mirey, Damien Dubois, Claude Petit, Eric Oswald, Jean-Philippe Nougayrède
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	DNA cross-linking agents DNA damage DNA damage checkpoints Escherichia coli Escherichia toxins genotoxicity Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/2f0d926a50d440af8b39cd1eca2c38c9
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:2f0d926a50d440af8b39cd1eca2c38c9
record_format	dspace
spelling	oai:doaj.org-article:2f0d926a50d440af8b39cd1eca2c38c92021-11-15T15:53:27ZThe Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells10.1128/mBio.02393-172150-7511https://doaj.org/article/2f0d926a50d440af8b39cd1eca2c38c92018-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02393-17https://doaj.org/toc/2150-7511ABSTRACT 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. IMPORTANCE Colorectal cancer is the third-most-common cause of cancer death. In addition to known risk factors such as high-fat diets and alcohol consumption, genotoxic intestinal Escherichia coli bacteria producing colibactin are proposed to play a role in colon cancer development. Here, by using transient infections with genotoxic E. coli, we showed that colibactins directly generate DNA cross-links in cellulo. Such lesions are converted into double-strand breaks during the repair response. DNA cross-links, akin to those induced by metabolites of alcohol and high-fat diets and by widely used anticancer drugs, are both severely mutagenic and profoundly cytotoxic lesions. This finding of a direct induction of DNA cross-links by a bacterium should facilitate delineating the role of E. coli in colon cancer and engineering new anticancer agents.Nadège Bossuet-GreifJulien VignardFrédéric TaiebGladys MireyDamien DuboisClaude PetitEric OswaldJean-Philippe NougayrèdeAmerican Society for MicrobiologyarticleDNA cross-linking agentsDNA damageDNA damage checkpointsEscherichia coliEscherichia toxinsgenotoxicityMicrobiologyQR1-502ENmBio, Vol 9, Iss 2 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	DNA cross-linking agents DNA damage DNA damage checkpoints Escherichia coli Escherichia toxins genotoxicity Microbiology QR1-502
spellingShingle	DNA cross-linking agents DNA damage DNA damage checkpoints Escherichia coli Escherichia toxins genotoxicity Microbiology QR1-502 Nadège Bossuet-Greif Julien Vignard Frédéric Taieb Gladys Mirey Damien Dubois Claude Petit Eric Oswald Jean-Philippe Nougayrède The Colibactin Genotoxin Generates DNA Interstrand Cross-Links in Infected Cells
description	ABSTRACT 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. IMPORTANCE Colorectal cancer is the third-most-common cause of cancer death. In addition to known risk factors such as high-fat diets and alcohol consumption, genotoxic intestinal Escherichia coli bacteria producing colibactin are proposed to play a role in colon cancer development. Here, by using transient infections with genotoxic E. coli, we showed that colibactins directly generate DNA cross-links in cellulo. Such lesions are converted into double-strand breaks during the repair response. DNA cross-links, akin to those induced by metabolites of alcohol and high-fat diets and by widely used anticancer drugs, are both severely mutagenic and profoundly cytotoxic lesions. This finding of a direct induction of DNA cross-links by a bacterium should facilitate delineating the role of E. coli in colon cancer and engineering new anticancer agents.
format	article
author	Nadège Bossuet-Greif Julien Vignard Frédéric Taieb Gladys Mirey Damien Dubois Claude Petit Eric Oswald Jean-Philippe Nougayrède
author_facet	Nadège Bossuet-Greif Julien Vignard Frédéric Taieb Gladys Mirey Damien Dubois Claude Petit Eric Oswald Jean-Philippe Nougayrède
author_sort	Nadège Bossuet-Greif
title	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_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_sort	colibactin genotoxin generates dna interstrand cross-links in infected cells
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/2f0d926a50d440af8b39cd1eca2c38c9
work_keys_str_mv	AT nadegebossuetgreif thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT julienvignard thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT frederictaieb thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT gladysmirey thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT damiendubois thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT claudepetit thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT ericoswald thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT jeanphilippenougayrede thecolibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT nadegebossuetgreif colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT julienvignard colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT frederictaieb colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT gladysmirey colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT damiendubois colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT claudepetit colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT ericoswald colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells AT jeanphilippenougayrede colibactingenotoxingeneratesdnainterstrandcrosslinksininfectedcells
_version_	1718427265501495296

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

Ejemplares similares