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Infection and Immunity, April 2000, p. 1919-1927, Vol. 68, No. 4
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Bordetella pertussis TonB, a Bvg-Independent Virulence Determinant

Elizabeth Pradel,1 Nicole Guiso,2 Franco D. Menozzi,1 and Camille Locht1,*

INSERM U447, Institut Pasteur de Lille, 59019 Lille Cedex,1 and Laboratoire des Bordetella, Institut Pasteur, 75724 Paris Cedex 15,2 France

Received 23 August 1999/Returned for modification 21 October 1999/Accepted 20 December 1999

In gram-negative bacteria, high-affinity iron uptake requires the TonB/ExbB/ExbD envelope complex to release iron chelates from their specific outer membrane receptors into the periplasm. Based on sequence similarities, the Bordetella pertussis tonB exbB exbD locus was identified on a cloned DNA fragment. The tight organization of the three genes suggests that they are cotranscribed. A putative Fur-binding sequence located upstream from tonB was detected in a Fur titration assay, indicating that the tonB exbB exbD operon may be Fur-repressed in high-iron growth conditions. Putative structural genes of the beta -subunit of the histone-like protein HU and of a new two-component regulatory system were identified upstream from tonB and downstream from exbD, respectively. A B. pertussis Delta tonB exbB::Kmr mutant was constructed by allelic exchange and characterized. The mutant was impaired for growth in low-iron medium in vitro and could not use ferrichrome, desferal, or hemin as iron sources. Levels of production of the major bacterial toxins and adhesins were similar in the TonB+/TonB- pair. The Delta tonB exbB mutant was still responsive to chemical modulators of virulence; thus, the BvgA/BvgS two-component system is not TonB dependent. Nevertheless, in vivo in the mouse respiratory infection model, the colonization ability of the mutant was reduced compared to the parental strain.

* Corresponding author. Mailing address: INSERM U447, Institut Pasteur de Lille, 1 rue du Prof. Calmette, BP 245, 59019 Lille Cedex, France. Phone: 33 (0) 3-20-87-11-51. Fax: 33 (0) 3-20-87-11-58. E-mail: Camille.Locht{at}pasteur-lille.fr.

Infection and Immunity, April 2000, p. 1919-1927, Vol. 68, No. 4
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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