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Infection and Immunity, October 1999, p. 5091-5099, Vol. 67, No. 10
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The High-Pathogenicity Island of Yersinia enterocolitica Ye8081 Undergoes Low-Frequency Deletion but Not Precise Excision, Suggesting Recent Stabilization in the Genome

Sandrine Bach,1 Carmen Buchrieser,1 Michael Prentice,1,2 Annie Guiyoule,1 Tarek Msadek,3 and Elisabeth Carniel1,*

Unité de Bactériologie Moléculaire et Médicale, Laboratoire des Yersinia,1 and Unité de Biochimie Microbienne,3 Institut Pasteur, 75724 Paris Cedex 15, France, and Department of Medical Microbiology, St. Bartholomew's and the Royal London School of Medicine and Dentistry, London EC1A 7BE, United Kingdom2

Received 18 May 1999/Returned for modification 24 June 1999/Accepted 27 July 1999

Highly pathogenic strains of Yersinia pestis, Y. pseudotuberculosis, and Y. enterocolitica are characterized by the possession of a pathogenicity island designated the high-pathogenicity island (HPI). This 35- to 45-kb island carries an iron uptake system named the yersiniabactin locus. While the HPIs of Y. pestis and Y. pseudotuberculosis are subject to high-frequency spontaneous deletion from the chromosome, we were initially unable to obtain HPI-deleted Y. enterocolitica 1B isolates. In the present study, using a positive selection strategy, we identified three HPI-deleted mutants of Y. enterocolitica strain Ye8081. In these three independent clones, the chromosomal deletion was not limited to the HPI but encompassed a larger DNA fragment of approximately 140 kb. Loss of this fragment, which occurred at a frequency of approximately 5 × 10-7, resulted in the disappearance of several phenotypic traits, such as growth in a minimal medium, hydrolysis of o-nitrophenyl-beta -D-thiogalactopyranoside, Tween esterase activity, and motility, and in a decreased virulence for mice. However, no precise excision of the Ye8081 HPI was observed. To gain more insight into the molecular basis for this phenomenon, the putative machinery of HPI excision in Y. enterocolitica was analyzed and compared to that in Y. pseudotuberculosis. We show that the probable reasons for failure of precise excision of the HPI of Y. enterocolitica Ye8081 are (i) the interruption of the P4-like integrase gene located close to its right-hand boundary by a premature stop codon and (ii) lack of conservation of 17-bp att-like sequences at both extremities of the HPI. These mutations may represent a process of HPI stabilization in the species Y. enterocolitica.

* Corresponding author. Mailing address: Institut Pasteur, Unité de Bactériologie Moléculaire et Médicale, Laboratoire des Yersinia, 28, rue du Dr. Roux, 75724 Paris Cedex 15, France. Phone: (33-1)-45-68-83-26. Fax: (33-1)-40-61-30-01. E-mail: carniel2{at}pasteur.fr.

Infection and Immunity, October 1999, p. 5091-5099, Vol. 67, No. 10
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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