title: Paralysis and killing of Caenorhabditis elegans by enteropathogenic Escherichia coli requires the bacterial tryptophanase gene.
abstract: Pathogenic Escherichia coli, including enteropathogenic E. coli (EPEC), enterohaemorrhagic E. coli (EHEC), enteroinvasive E. coli (EIEC) and enterotoxigenic E. coli (ETEC) are major causes of food and water-borne disease. We have developed a genetically tractable model of pathogenic E. coli virulence based on our observation that these bacteria paralyse and kill the nematode Caenorhabditis elegans. Paralysis and killing of C. elegans by EPEC did not require direct contact, suggesting that a secreted toxin mediates the effect. Virulence against C. elegans required tryptophan and bacterial tryptophanase, the enzyme catalysing the production of indole and other molecules from tryptophan. Thus, lack of tryptophan in growth media or deletion of tryptophanase gene failed to paralyse or kill C. elegans. While known tryptophan metabolites failed to complement an EPEC tryptophanase mutant when presented extracellularly, complementation was achieved with the enzyme itself expressed either within the pathogen or within a cocultured K12 strains. Thus, an unknown metabolite of tryptophanase, derived from EPEC or from commensal non-pathogenic strains, appears to directly or indirectly regulate toxin production within EPEC. EPEC strains containing mutations in the locus of enterocyte effacement (LEE), a pathogenicity island required for virulence in humans, also displayed attenuated capacity to paralyse and kill nematodes. Furthermore, tryptophanase activity was required for full activation of the LEE1 promoter, and for efficient formation of actin-filled membranous protrusions (attaching and effacing lesions) that form on the surface of mammalian epithelial cells following attachment and which depends on LEE genes. Finally, several C. elegans genes, including hif-1 and egl-9, rendered C. elegans less susceptible to EPEC when mutated, suggesting their involvement in mediating toxin effects. Other genes including sek-1, mek-1, mev-1, pgp-1,3 and vhl-1, rendered C. elegans more susceptible to EPEC effects when mutated, suggesting their involvement in protecting the worms. Moreover we have found that C. elegans genes controlling lifespan (daf-2, age-1 and daf-16), also mediate susceptibility to EPEC. Together, these data suggest that this C. elegans/EPEC system will be valuable in elucidating novel factors relevant to human disease that regulate virulence in the pathogen or susceptibility to infection in the host.
authors.affiliation: Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA, USA.
authors.forename: Akwasi
authors.initials: A
authors.lastname: Anyanful
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Dolan-Livengood
authors.forename: Taiesha
authors.initials: T
authors.lastname: Lewis
authors.forename: Seema
authors.initials: S
authors.lastname: Sheth
authors.forename: Mark N
authors.initials: MN
authors.lastname: Dezalia
authors.forename: Melanie A
authors.initials: MA
authors.lastname: Sherman
authors.forename: Lisa V
authors.initials: LV
authors.lastname: Kalman
authors.forename: Guy M
authors.initials: GM
authors.lastname: Benian
authors.forename: Daniel
authors.initials: D
authors.lastname: Kalman
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics* / Q000378:metabolism; D001692:Biological Transport; D017173:Caenorhabditis elegans / Q000235:genetics / Q000378:metabolism / Q000382:microbiology*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D007211:Indoles / Q000494:pharmacology; D009154:Mutation; D010750:Phosphoproteins / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects; D014364:Tryptophan / Q000378:metabolism / Q000494:pharmacology; D014368:Tryptophanase / Q000235:genetics* / Q000378:metabolism; D014774:Virulence
chemical_list: D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D007211:Indoles; C457340:LEE protein, E coli; D010750:Phosphoproteins; C030374:indole; D014364:Tryptophan; D014368:Tryptophanase