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

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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 IdeR

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 aputative formyltransferase, an essential enzyme in the exochelinbiosynthetic pathway (E. H. Fiss, Y. Yu, and W. R. Jacobs, Jr.,Mol. Microbiol. 14:557-569, 1994). We investigated the regulationof fxbA by the mycobacterial IdeR, a homolog of the Corynebacteriumdiphtheriae 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 tothe fxbA regulatory region in the presence of divalent metals.DNase I footprinting assays indicated that IdeR binding protectsa 28-bp region containing a palindromic sequence of the fxbA promoterthat was identified in primer extension assays. fxbA regulationwas measured in M. smegmatis wild-type and ideR mutant strainscontaining fxbA promoter-lacZ fusions. These experiments confirmedthat fxbA expression is negatively regulated by iron and showedthat inactivation of ideR results in iron-independent expressionof fxbA. However, the levels of its expression in the ideR mutantwere approximately 50% lower than those in the wild-type strainunder iron limitation, indicating an undefined positive role ofIdeR 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.

Publication no. 64 from the TB Center, Public Health ResearchInstitute.

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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