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Microbiology 154 (2008), 1763-1774; DOI  10.1099/mic.0.2007/015537-0
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Microbiology 154 (2008), 1763-1774; DOI  10.1099/mic.0.2007/015537-0
© 2008 Society for General Microbiology

Pleiotropic roles of iron-responsive transcriptional regulator Fur in Burkholderia multivorans

Satoshi Yuhara, Harunobu Komatsu, Hiroyuki Goto, Yoshiyuki Ohtsubo, Yuji Nagata and Masataka Tsuda

Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Sendai 980-8577, Japan

Correspondence
Masataka Tsuda
mtsuda{at}ige.tohoku.ac.jp

The fur (ferric uptake regulator) gene of Burkholderia multivorans ATCC 17616 was identified by transposon mutagenesis analysis. The fur deletion mutant of strain ATCC 17616 (i) constitutively produced siderophores, (ii) was more sensitive to reactive oxygen species (ROS) than the wild-type strain, (iii) showed lower superoxide dismutase and catalase activities than the wild-type strain, (iv) was unable to grow on M9 minimal agar plates containing several substrates that can be used as sole carbon sources by the wild-type strain, and (v) was hypersensitive to nitrite and nitric oxide under microaerobic and aerobic conditions, respectively. These results clearly indicate that the Fur protein in strain ATCC 17616 plays pleiotropic roles in iron homeostasis, removal and/or resistance to ROS and nitrosative stress, and energy metabolism. Furthermore, employment of an in vivo Fur titration assay system led to the isolation from the ATCC 17616 genome of 13 Fur-binding DNA regions, and a subsequent electrophoretic mobility-shift assay confirmed the direct binding of Fur protein to all of these DNA regions. Transcriptional analysis of the genes located just downstream of the Fur-binding sites demonstrated that Fur acts as a repressor for these genes. Nine of the 13 regions were presumed to be involved in the acquisition and utilization of iron.

Abbreviations: CAS, Chrome Azurol S; (P)EB, (Poor) Environmental Bacteria (medium); ECF, extracytoplasmic function; EMSA; electrophoretic mobility-shift assay; Fur, ferric uptake regulator; FURTA, Fur titration assay; qRT-PCR, quantitative real-time reverse-transcription PCR; ROS, reactive oxygen species; SOD, superoxide dismutase

The GenBank/EMBL/DDBJ accession numbers for the sequences reported in this paper are AP009835–AP009838.

Two supplementary tables with details of the PCR primer sequences and gene annotation are available with the online version of this paper.






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