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Infection and Immunity, March 2000, p. 1164-1170, Vol. 68, No. 3
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Actinobacillus pleuropneumoniae Iron Transport: a Set of exbBD Genes Is Transcriptionally Linked to the tbpB Gene and Required for Utilization of Transferrin-Bound Iron

Walaiporn Tonpitak, Svenja Thiede, Winfried Oswald, Nina Baltes, and Gerald-F. Gerlach*

Institut für Mikrobiologie und Tierseuchen, Tierärztliche Hochschule Hannover, 30173 Hannover, Germany

Received 3 September 1999/Returned for modification 3 November 1999/Accepted 29 November 1999

Upon iron restriction, Actinobacillus pleuropneumoniae has been shown to express the transferrin-binding proteins TbpB and TbpA, both of which have been implied to be important virulence factors. In order to identify additional iron-regulated proteins, we cloned and analyzed the region upstream of the transferrin-binding protein genes in an A. pleuropneumoniae serotype 7 strain. We located immediately upstream of the tbpB gene two open reading frames which were 43% homologous to the neisserial ExbBD protein genes. By raising specific antibodies, we showed that ExbB is expressed under iron-limiting growth conditions only, and RT-PCR analysis revealed that the exbBD genes and the tbpB gene are transcribed on a single polycistronic mRNA. By constructing an isogenic and nonpolar exbBD mutant, we showed that the exbBD genes are required by A. pleuropneumoniae for utilization of transferrin-bound iron. Using PCR and Western blotting, we showed that the genetic organization found in A. pleuropneumoniae serotype 7 is similar in all 12 A. pleuropneumoniae serotype reference strains.

* Corresponding author. Mailing address: Institut für Mikrobiologie und Tierseuchen, Tierärztliche Hochschule Hannover, Bischofsholer Damm 15, 30173 Hannover, Germany. Phone: 49-511-856 7598. Fax: 49-511-856 7697. E-mail: ggerlach{at}micro.tiho-hannover.de.

Infection and Immunity, March 2000, p. 1164-1170, Vol. 68, No. 3
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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