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Journal of Bacteriology, May 2006, p. 3631-3644, Vol. 188, No. 10
0021-9193/06/$08.00+0     doi:10.1128/JB.188.10.3631-3644.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Ornibactin Biosynthesis and Transport Genes of Burkholderia cenocepacia Are Regulated by an Extracytoplasmic Function {sigma} Factor Which Is a Part of the Fur Regulon{dagger}

Kirsty Agnoli, Carolyn A. Lowe,{ddagger} Kate L. Farmer, Seyyed I. Husnain, and Mark S. Thomas*

Division of Genomic Medicine, School of Medicine and Biomedical Sciences, University of Sheffield, Sheffield S10 2RX, United Kingdom

Received 16 December 2005/ Accepted 13 February 2006

Burkholderia cenocepacia mutants that fail to produce the siderophore ornibactin were obtained following mutagenesis with mini-Tn5Tp. These mutants were shown to be growth restricted under conditions of iron depletion. In eight of the mutants, the transposon had integrated into one of two genes, orbI and orbJ, encoding nonribosomal peptide synthetases. In the other mutant, the transposon had inserted into an open reading frame, orbS, located upstream from orbI. The polypeptide product of orbS exhibits a high degree of similarity to the Pseudomonas aeruginosa extracytoplasmic function (ECF) {sigma} factor PvdS but possesses an N-terminal extension of approximately 29 amino acids that is not present in PvdS. Three predicted OrbS-dependent promoters were identified within the ornibactin gene cluster, based on their similarity to PvdS-dependent promoters. The iron-regulated activity of these promoters was shown to require OrbS. Transcription of the orbS gene was found to be under the control of an iron-regulated {sigma}70-dependent promoter. This promoter, but not the OrbS-dependent promoters, was shown to be a target for repression by the global regulator Fur. Our results demonstrate that production of ornibactin by B. cenocepacia in response to iron starvation requires transcription of an operon that is dependent on the Fur-regulated ECF {sigma} factor gene orbS. A mechanism is also proposed for the biosynthesis of ornibactin.

* Corresponding author. Mailing address: Division of Genomic Medicine, School of Medicine and Biomedical Sciences, University of Sheffield, Beech Hill Road, Sheffield S10 2RX, United Kingdom. Phone: 44 114 2712834. Fax: 44 114 2713892. E-mail: m.s.thomas{at}shef.ac.uk.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

{ddagger} Present address: Department of Immunology, Strathclyde Institute for Biomedical Sciences, University of Strathclyde, Glasgow G4 0NR, United Kingdom.

Journal of Bacteriology, May 2006, p. 3631-3644, Vol. 188, No. 10
0021-9193/06/$08.00+0     doi:10.1128/JB.188.10.3631-3644.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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