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Infection and Immunity, March 2009, p. 1155-1164, Vol. 77, No. 3
0019-9567/09/$08.00+0     doi:10.1128/IAI.01082-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Roles of the Extraintestinal Pathogenic Escherichia coli ZnuACB and ZupT Zinc Transporters during Urinary Tract Infection{triangledown}

Mourad Sabri, Sébastien Houle, and Charles M. Dozois*

INRS-Institut Armand-Frappier, Laval, Québec, Canada

Received 29 August 2008/ Returned for modification 13 October 2008/ Accepted 15 December 2008

Roles of the ZnuACB and ZupT transporters were assessed in Escherichia coli K-12 and uropathogenic E. coli (UPEC) CFT073. K-12 and CFT073 {Delta}znu {Delta}zupT mutants demonstrated decreased 65Zn2+ uptake and growth in minimal medium. CFT073{Delta}znu demonstrated an intermediate decrease of 65Zn2+ uptake and growth in minimal medium, whereas the CFT073{Delta}zupT mutant grew as well as CFT073 and exhibited a less marked decrease in 65Zn2+ uptake. CFT073 mutants grew as well as the wild type in human urine. In competitive infections in CBA/J mice, the {Delta}zupT mutant demonstrated no disadvantage during urinary tract infection. In contrast, the UPEC {Delta}znu and {Delta}znu {Delta}zupT strains demonstrated significantly reduced numbers in the bladders (mean 4.4- and 30-fold reductions, respectively) and kidneys (mean 41- and 48-fold reductions, respectively). In addition, in single-strain infection experiments, the {Delta}znu and {Delta}znu {Delta}zupT mutants were reduced in the kidneys (P = 0.0012 and P < 0.0001, respectively). Complementation of the CFT073 {Delta}znu {Delta}zupT mutant with the znuACB genes restored growth in Zn-deficient medium and bacterial numbers in the bladder and kidneys. The loss of the zinc transport systems decreased both motility and resistance to hydrogen peroxide, which could be restored by supplementation with zinc. Overall, the results indicate that Znu and ZupT are required for growth in zinc limited-conditions, that Znu is the predominant zinc transporter, and that the loss of Znu and ZupT has a cumulative effect on fitness during UTI, which may in part be due to reduced resistance to oxidative stress and motility.

* Corresponding author. Mailing address: INRS-Institut Armand-Frappier, 531 Blvd. des Prairies, Laval, Québec H7V 1B7, Canada. Phone: (450) 687-5010, x4221. Fax: (450) 686-5501. E-mail: charles.dozois{at}iaf.inrs.ca

{triangledown} Published ahead of print on 22 December 2008.

Editor: B. A. McCormick

Infection and Immunity, March 2009, p. 1155-1164, Vol. 77, No. 3
0019-9567/09/$08.00+0     doi:10.1128/IAI.01082-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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