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Journal of Bacteriology, February 2004, p. 938-948, Vol. 186, No. 4
0021-9193/04/$08.00+0 DOI: 10.1128/JB.186.4.938-948.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.
and Sandra K. Armstrong*
Department of Microbiology, University of Minnesota Medical School, Minneapolis, Minnesota 55455-0312
Received 28 July 2003/ Accepted 5 November 2003
The Bordetella pertussis heme utilization gene cluster hurIR bhuRSTUV encodes regulatory and transport functions required for assimilation of iron from heme and hemoproteins. Expression of the bhu genes is iron regulated and heme inducible. The putative extracytoplasmic function (ECF)
factor, HurI, is required for heme-responsive bhu gene expression. In this study, transcriptional activation of B. pertussis bhu genes in response to heme compounds was shown to be dose dependent and specific for heme; protoporphyrin IX and other heme structural analogs did not activate bhu gene expression. Two promoters controlling expression of the heme utilization genes were mapped by primer extension analysis. The hurI promoter showed similarity to
70-like promoters, and its transcriptional activity was iron regulated and heme independent. A second promoter identified upstream of bhuR exhibited little similarity to previously characterized ECF
factor-dependent promoters. Expression of bhuR was iron regulated, heme responsive, and hurI dependent in B. pertussis, as shown in a previous study with Bordetella bronchiseptica. Further analyses showed that transcription originating at a distal upstream site and reading through the hurR-bhuR intergenic region contributes to bhuR expression under iron starvation conditions in the absence of heme inducer. The pattern of regulation of the readthrough transcript was consistent with transcription from the hurI promoter. The positions and regulation of the two promoters within the hur-bhu gene cluster influence the production of heme transport machinery so that maximal expression of the bhu genes occurs under iron starvation conditions only in the presence of heme iron sources.
Present address: Laboratory of Molecular Biology, National Cancer Institute, Bethesda, MD 20892-4264.
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