colibactin and Disease-Models--Animal

SignificanceIn a polymicrobial battlefield where different species compete for nutrients and colonization niches, antimicrobial compounds are the sword and shield of commensal microbes in competition with invading pathogens and each other. The identification of an

Topics: Animals; Antibiosis; Cholera; Disease Models, Animal; DNA Damage; Escherichia coli; Gastrointestinal Microbiome; Host-Pathogen Interactions; Humans; Mice; Microbial Interactions; Mutagens; Peptides; Polyketides; Prognosis; Reactive Oxygen Species; Vibrio cholerae

Colibactin is synthesized by a 54-kb genomic island, leads to toxicity in eukaryotic cells, and plays a vital role in many diseases, including neonatal sepsis and meningitis. Avian pathogenic

Topics: Animals; Disease Models, Animal; Escherichia coli; Escherichia coli Infections; Escherichia coli Proteins; Meningitis; Mice; Peptides; Polyketides; RNA, Messenger

Colibactin is a complex secondary metabolite that leads to genotoxicity that interferes with the eukaryotic cell cycle. It plays an important role in many diseases, including neonatal mouse sepsis and meningitis. Avian pathogenic

Topics: Animals; Brain; Cell Line; Cytokines; Disease Models, Animal; Escherichia coli; Escherichia coli Infections; Female; Male; Mice, Inbred ICR; Peptides; Polyketides; Poultry Diseases; Tight Junction Proteins

The genotoxin colibactin, synthesized by Escherichia coli, is a secondary metabolite belonging to the chemical family of hybrid polyketide/nonribosomal peptide compounds. It is produced by a complex biosynthetic assembly line encoded by the pks pathogenicity island. The presence of this large cluster of genes in the E. coli genome is invariably associated with the high-pathogenicity island, encoding the siderophore yersiniabactin, which belongs to the same chemical family as colibactin. The E. coli heat shock protein HtpG (Hsp90Ec) is the bacterial homolog of the eukaryotic molecular chaperone Hsp90, which is involved in the protection of cellular proteins against a variety of environmental stresses. In contrast to eukaryotic Hsp90, the functions and client proteins of Hsp90Ec are poorly known. Here, we demonstrated that production of colibactin and yersiniabactin is abolished in the absence of Hsp90Ec We further characterized an interplay between the Hsp90Ec molecular chaperone and the ClpQ protease involved in colibactin and yersiniabactin synthesis. Finally, we demonstrated that Hsp90Ec is required for the full in vivo virulence of extraintestinal pathogenic E. coli This is the first report highlighting the role of heat shock protein Hps90Ec in the production of two secondary metabolites involved in E. coli virulence.

Topics: Animals; Disease Models, Animal; Endopeptidase Clp; Escherichia coli; Escherichia coli Infections; Escherichia coli Proteins; Female; Gene Deletion; HSP90 Heat-Shock Proteins; Mice, Inbred C57BL; Mutagens; Peptides; Phenols; Polyketides; Protein Interaction Mapping; Rats, Wistar; Siderophores; Thiazoles; Virulence

Various forms of cancer have been linked to the carcinogenic activities of microorganisms(1-3). The virulent gene island polyketide synthase (pks) produces the secondary metabolite colibactin, a genotoxic molecule(s) causing double-stranded DNA breaks(4) and enhanced colorectal cancer development(5,6). Colibactin biosynthesis involves a prodrug resistance strategy where an N-terminal prodrug scaffold (precolibactin) is assembled, transported into the periplasm and cleaved to release the mature product(7-10). Here, we show that ClbM, a multidrug and toxic compound extrusion (MATE) transporter, is a key component involved in colibactin activity and transport. Disruption of clbM attenuated pks+ E. coli-induced DNA damage in vitro and significantly decreased the DNA damage response in gnotobiotic Il10(-/-) mice. Colonization experiments performed in mice or zebrafish animal models indicate that clbM is not implicated in E. coli niche establishment. The X-ray structure of ClbM shows a structural motif common to the recently described MATE family. The 12-transmembrane ClbM is characterized as a cation-coupled antiporter, and residues important to the cation-binding site are identified. Our data identify ClbM as a precolibactin transporter and provide the first structure of a MATE transporter with a defined and specific biological function.

Topics: Animals; Crystallography, X-Ray; Disease Models, Animal; DNA Damage; Escherichia coli; Escherichia coli Infections; Escherichia coli Proteins; Ilex; Mice; Models, Molecular; Mutagens; Organic Cation Transport Proteins; Peptides; Polyketides; Protein Conformation; Protein Transport; Zebrafish

Escherichia coli strains expressing the K1 capsule are a major cause of sepsis and meningitis in human neonates. The development of these diseases is dependent on the expression of a range of virulence factors, many of which remain uncharacterized. Here, we show that all but 1 of 34 E. coli K1 neonatal isolates carried clbA and clbP, genes contained within the pks pathogenicity island and required for the synthesis of colibactin, a polyketide-peptide genotoxin that causes genomic instability in eukaryotic cells by induction of double-strand breaks in DNA. Inactivation of clbA and clbP in E. coli A192PP, a virulent strain of serotype O18:K1 that colonizes the gastrointestinal tract and translocates to the blood compartment with very high frequency in experimental infection of the neonatal rat, significantly reduced the capacity of A192PP to colonize the gut, engender double-strand breaks in DNA, and cause invasive, lethal disease. Mutation of clbA, which encodes a pleiotropic enzyme also involved in siderophore synthesis, impacted virulence to a greater extent than mutation of clbP, encoding an enzyme specific to colibactin synthesis. Restoration of colibactin gene function by complementation reestablished the fully virulent phenotype. We conclude that colibactin contributes to the capacity of E. coli K1 to colonize the neonatal gastrointestinal tract and to cause invasive disease in the susceptible neonate.

Topics: Animals; Animals, Newborn; Base Sequence; Disease Models, Animal; Escherichia coli; Escherichia coli Infections; Genomic Islands; Immunohistochemistry; Molecular Sequence Data; Peptides; Polyketides; Polymerase Chain Reaction; Rats; Rats, Wistar; Virulence