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Infection and Immunity, October 1999, p. 5060-5068, Vol. 67, No. 10
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification of the znuA-Encoded Periplasmic Zinc Transport Protein of Haemophilus ducreyi

David A. Lewis,1,2 Julia Klesney-Tait,1 Sheryl R. Lumbley,1 Christine K. Ward,1 Jo L. Latimer,1 Catherine A. Ison,3 and Eric J. Hansen1,*

Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas 75235-9048,1 and Department of Genitourinary Medicine and Communicable Disease2 and Department of Microbiology,3 Imperial College School of Medicine, St. Mary's Campus, Norfolk Place, London W2 1PG, United Kingdom

Received 28 April 1999/Returned for modification 4 June 1999/Accepted 6 July 1999

The znuA gene of Haemophilus ducreyi encodes a 32-kDa (mature) protein that has homology to both the ZnuA protein of Escherichia coli and the Pzp1 protein of H. influenzae; both of these latter proteins are members of a growing family of prokaryotic zinc transporters. Inactivation of the H. ducreyi 35000 znuA gene by insertional mutagenesis resulted in a mutant that grew more slowly than the wild-type parent strain in vitro unless ZnCl2 was provided at a final concentration of 100 µM. Other cations tested did not restore growth of this H. ducreyi mutant to wild-type levels. The H. ducreyi ZnuA protein was localized to the periplasm, where it is believed to function as the binding component of a zinc transport system. Complementation of the znuA mutation with the wild-type H. ducreyi znuA gene provided in trans restored the ability of this H. ducreyi mutant to grow normally in the absence of exogenously added ZnCl2. The wild-type H. ducreyi znuA gene was also able to complement a H. influenzae pzp1 mutation. The H. ducreyi znuA isogenic mutant exhibited significantly decreased virulence (P = 0.0001) when tested in the temperature-dependent rabbit model for experimental chancroid. This decreased virulence was not observed when the znuA mutant was complemented with the wild-type H. ducreyi znuA gene provided in trans.

* Corresponding author. Mailing address: Department of Microbiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75235-9048. Phone: (214) 648-5974. Fax: (214) 648-5905. E-mail: hansen01{at}utsw.swmed.edu.

Infection and Immunity, October 1999, p. 5060-5068, Vol. 67, No. 10
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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