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Journal of Bacteriology, July 2001, p. 4278-4287, Vol. 183, No. 14
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.14.4278-4287.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
The Bordetella bhu Locus Is Required
for Heme Iron Utilization
Carin K.
Vanderpool and
Sandra K.
Armstrong*
Department of Microbiology, University of
Minnesota Medical School, Minneapolis, Minnesota 55455-0312
Received 8 March 2001/Accepted 24 April 2001
Bordetella pertussis and Bordetella
bronchiseptica are capable of obtaining iron from hemin and
hemoglobin. Genes encoding a putative bacterial heme iron acquisition
system (bhu, for Bordetella heme
utilization) were identified in a B. pertussis genomic
sequence database, and the corresponding DNA was isolated from a
virulent strain of B. pertussis. A B. pertussis
bhuR mutant, predicted to lack the heme outer membrane
receptor, was generated by allelic exchange. In contrast to the
wild-type strain, bhuR mutant PM5 was incapable of
acquiring iron from hemin and hemoglobin; genetic complementation of
PM5 with the cloned bhuRSTUV genes restored heme
utilization to wild-type levels. In parallel studies, B. bronchiseptica bhu sequences were also identified and a
B. bronchiseptica bhuR mutant was constructed and
confirmed to be defective in heme iron acquisition. The wild-type
B. bronchiseptica parent strain grown under low-iron
conditions produced the presumptive BhuR protein, which was absent in
the bhuR mutant. Furthermore, production of BhuR by
iron-starved B. bronchiseptica was markedly enhanced by
culture in hemin-supplemented medium, suggesting that these organisms
sense and respond to heme in the environment. Analysis of the genetic
region upstream of the bhu cluster identified open reading frames predicted to encode homologs of the Escherichia coli ferric citrate uptake regulators FecI and FecR.
These putative Bordetella regulators may mediate
heme-responsive positive transcriptional control of the
bhu genes.
*
Corresponding author. Mailing address: Department of
Microbiology, University of Minnesota, MMC 196 FUMC, 420 Delaware St. S.E., Minneapolis, MN 55455-0312. Phone: (612) 625-6947. Fax: (612)
626-0623. E-mail: sandra{at}lenti.med.umn.edu.
Journal of Bacteriology, July 2001, p. 4278-4287, Vol. 183, No. 14
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.14.4278-4287.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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