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Journal of Bacteriology, June 1999, p. 3402-3408, Vol. 181, No. 11
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Transcriptional Control of the Iron-Responsive fxbA Gene by the Mycobacterial Regulator IdeRdagger

Olivier Dussurget,1,2 Juliano Timm,1 Manuel Gomez,1 Benjamin Gold,1,3 Shengwei Yu,4 Sue Z. Sabol,5 Randall K. Holmes,6 William R. Jacobs Jr.,4 and Issar Smith1,*

TB Center, Public Health Research Institute,1 and Department of Microbiology, New York University Medical Center,3 New York, New York 10016; UFR de Biochimie, Université Paris 7, 75251 Paris Cedex 05, France2; Department of Microbiology and Immunology, Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, New York 104614; Section of Gene Structure and Regulation, Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 208925; and Department of Microbiology, University of Colorado Health Sciences Center, Denver, Colorado 802626

Received 12 March 1998/Accepted 30 March 1999

Exochelin is the primary extracellular siderophore of Mycobacterium smegmatis, and the iron-regulated fxbA gene encodes a putative formyltransferase, an essential enzyme in the exochelin biosynthetic pathway (E. H. Fiss, Y. Yu, and W. R. Jacobs, Jr., Mol. Microbiol. 14:557-569, 1994). We investigated the regulation of fxbA by the mycobacterial IdeR, a homolog of the Corynebacterium diphtheriae iron regulator DtxR (M. P. Schmitt, M. Predich, L. Doukhan, I. Smith, and R. K. Holmes, Infect. Immun. 63:4284-4289, 1995). Gel mobility shift experiments showed that IdeR binds to the fxbA regulatory region in the presence of divalent metals. DNase I footprinting assays indicated that IdeR binding protects a 28-bp region containing a palindromic sequence of the fxbA promoter that was identified in primer extension assays. fxbA regulation was measured in M. smegmatis wild-type and ideR mutant strains containing fxbA promoter-lacZ fusions. These experiments confirmed that fxbA expression is negatively regulated by iron and showed that inactivation of ideR results in iron-independent expression of fxbA. However, the levels of its expression in the ideR mutant were approximately 50% lower than those in the wild-type strain under iron limitation, indicating an undefined positive role of IdeR in the regulation of fxbA.

* Corresponding author. Mailing address: TB Center, Public Health Research Institute, 455 First Ave., New York, NY 10016. Phone: (212) 578-0867. Fax: (212) 578-0804. E-mail: smitty{at}phri.nyu.edu.

dagger Publication no. 64 from the TB Center, Public Health Research Institute.

Journal of Bacteriology, June 1999, p. 3402-3408, Vol. 181, No. 11
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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