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Journal of Bacteriology, February 2006, p. 1567-1576, Vol. 188, No. 4
0021-9193/06/$08.00+0     doi:10.1128/JB.188.4.1567-1576.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Tetrapyrrole Biosynthesis in Rhodobacter capsulatus Is Transcriptionally Regulated by the Heme-Binding Regulatory Protein, HbrL

James L. Smart and Carl E. Bauer*

Department of Biology, Indiana University, Bloomington, Indiana 47405

Received 7 July 2005/ Accepted 21 November 2005

We demonstrate that the expression of hem genes in Rhodobacter capsulatus is transcriptionally repressed in response to the exogenous addition of heme. A high-copy suppressor screen for regulators of hem gene expression resulted in the identification of an LysR-type transcriptional regulator, called HbrL, that regulates hem promoters in response to the availability of heme. HbrL is shown to activate the expression of hemA and hemZ in the absence of exogenous hemin and repress hemB expression in the presence of exogenous hemin. Heterologously expressed HbrL apoprotein binds heme b and is purified with bound heme b when expressed in the presence of 5-aminolevulinic acid. Electrophoretic gel shift analysis demonstrated that HbrL binds the promoter region of hemA, hemB, and hemZ as well as its own promoter and that the presence of heme increases the binding affinity of HbrL to hemB.

* Corresponding author. Mailing address: Department of Biology, Indiana University, Bloomington, IN 47405. Phone: (812) 855-6595. Fax: (812) 855-4178. E-mail: cbauer{at}bio.indiana.edu.

Journal of Bacteriology, February 2006, p. 1567-1576, Vol. 188, No. 4
0021-9193/06/$08.00+0     doi:10.1128/JB.188.4.1567-1576.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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