title: Disruption of tonB in Bordetella bronchiseptica and Bordetella pertussis prevents utilization of ferric siderophores, haemin and haemoglobin as iron sources.
abstract: The Bordetella bronchiseptica tonB gene was cloned by detection of a chromosomal restriction fragment hybridizing with each of two degenerate oligonucleotides that corresponded to Pro-Glu and Pro-Lys repeats characteristic of known TonB proteins. The tonB(Bb) gene was situated upstream of exbB and exbD homologues and downstream of a putative Fur-regulated promoter. Hybridization results indicated that the tonB operon and flanking regions were highly conserved between B. bronchiseptica, Bordetella pertussis and Bordetella parapertussis. Disruption of tonB in B. bronchiseptica resulted in inability to grow in iron-limiting media, and inability to utilize alcaligin, enterobactin, ferrichrome, desferroxamine B, haemin and haemoglobin. Although it was not possible to inactivate tonB in a clinical B. pertussis isolate, tonB was disrupted in a laboratory B. pertussis strain previously selected for the ability to grow on Luria-Bertani medium. This B. pertussis tonB mutant shared a similar iron complex utilization deficient phenotype with the B. bronchiseptica tonB mutant. The B. bronchiseptica tonB operon present on a plasmid did not complement an Escherichia coli tonB mutant, but inefficient reconstitution of enterobactin utilization was observed in one fepA mutant harbouring plasmid copies of the B. pertussis fepA homologue and tonB(Bb) operon.
authors.affiliation: Centers for Disease Control and Prevention, Respiratory Diseases Branch, 1600 Clifton Rd, Mailstop C02, Atlanta, GA 30333, USA1.
authors.forename: Mary Lou
authors.initials: ML
authors.lastname: Nicholson
authors.affiliation: Centers for Disease Control and Prevention, Respiratory Diseases Branch, 1600 Clifton Rd, Mailstop C02, Atlanta, GA 30333, USA1.
authors.forename: Bernard
authors.initials: B
authors.lastname: Beall
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D015139:Blotting, Southern; D016950:Bordetella bronchiseptica / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D001886:Bordetella pertussis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D003227:Conjugation, Genetic; D004758:Enterobactin / Q000378:metabolism; D029968:Escherichia coli Proteins; D005291:Ferrichrome / Q000031:analogs & derivatives / Q000494:pharmacology; D017353:Gene Deletion; D006427:Hemin / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D007501:Iron / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009876:Operon; D017422:Sequence Analysis, DNA; D017262:Siderophores / Q000378:metabolism; D014169:Transformation, Bacterial
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C052874:ExbB protein, E coli; D006454:Hemoglobins; D008565:Membrane Proteins; D017262:Siderophores; C057616:tonB protein, Bacteria; C489181:tonB protein, E coli; C060790:exbD protein, E coli; D005291:Ferrichrome; D004758:Enterobactin; C015657:albomycin; D006427:Hemin; D007501:Iron