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Infection and Immunity, November 2001, p. 6707-6717, Vol. 69, No. 11
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.11.6707-6717.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification and Characterization of the Hemophore-Dependent Heme Acquisition System of Yersinia pestis

Maria-Silvia Rossi,1 Jacqueline D. Fetherston,2 Sylvie Létoffé,1 Elisabeth Carniel,3 Robert D. Perry,2 and Jean-Marc Ghigo1,*

Unité des Membranes Bactériennes Institut Pasteur (CNRS URA 2172)1 and Unité de Bactériologie Moléculaire et Médicale, Laboratoire des Yersinia,3 75724 Paris Cedex 15, France, and Department of Microbiology and Immunology, University of Kentucky, Lexington, Kentucky 40536-02982

Received 10 May 2001/Returned for modification 6 July 2001/Accepted 10 August 2001

Yersinia pestis possesses a heme-protein acquisition system (Hmu) that allows it to utilize heme and heme-protein complexes as the sole sources of iron. Analysis of the Y. pestis CO92 genomic sequence revealed a second heme-protein acquisition gene cluster that shares homology with the hemophore-dependent heme acquisition system (Has system) of Serratia marcescens. This locus consisted of the hasRyp receptor gene, the hasAyp hemophore gene, and genes encoding components of the HasAyp dedicated ABC transporter factor (hasDEyp), as well as a tonB homologue (hasByp). By using a reconstituted secretion system in Escherichia coli, we showed that HasAyp is a secreted heme-binding protein and that expression of HasAyp is iron regulated in E. coli. The use of a transcriptional reporter fusion showed that the hasRADEB promoter is Fur regulated and has increased activity at 37°C. Hemoglobin utilization via the Hasyp system was studied with both E. coli and Y. pestis, for which has and has hmu mutant strains were used. No contribution of the Has system to heme utilization was observed in either E. coli or Y. pestis under the conditions we tested. Previously it was shown that a deletion of the Hmu system had no effect on the virulence of Y. pestis in a mouse model of bubonic plague. An Hmu- Has- double mutant also retained full virulence in this model of infection. This report constitutes the first attempt to investigate the contribution of the hemophore-dependent heme acquisition system in bacterial pathogenicity.

* Corresponding author. Mailing address: Unité des Membranes Bactériennes Institut Pasteur (CNRS URA 2172), 25 rue du Dr. Roux, 75724 Paris CEDEX 15, France. Phone: (1) 33 0140 61 32 77. Fax: (1) 33 01 45 68 87 90. E-mail: jmghigo{at}pasteur.fr.

Infection and Immunity, November 2001, p. 6707-6717, Vol. 69, No. 11
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.11.6707-6717.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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