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Purification and analysis of YvmBHis-tag protein. (A) SDS-PAGE analysis of purified YvmBHis-tag protein from E. coli cells. (B) Fractionated chromatography on a Superdex 200 h/300 gel filtration column. The YvmBHis-tag protein eluted as a major peak with an apparent molecular mass of 45 ± 5 kDa. (PDF 164 kb)

The yvmA-yvmC-cypX locus from B. subtilis and B. licheniformis ATCC 14580. (A) For each gene product of B. subtilis and its equivalent in B. licheniformis ATCC 14580 a percentage of identity is indicated. The position of the genes on each chromosome is given. The yvmC and cypX genes encoding the enzymes involved in pulcherriminic synthesis are represented by purple arrows, the MarR-type regulators YvmB by brown arrows, and the MFS-like transporters YvmA by white arrows. The conserved palindromic motif, palindrome I, is indicated by blue boxes. (B) Alignment of the 42-bp palindromic motifs from the promoter regions of B. subtilis yvmB and B. licheniformis yvmA genes. Stars indicate the seven nucleotides, which are not conserved between the two palindromic sequences. The 14Â bp core region highly conserved in the two sequences is boxed. (PDF 147 kb)

Heatmap representation of proteins that were found differentially abundant between the Î yvmB mutant and the wild type strain in the cytosolic fraction. Heatmap shows the change in protein levels among four independent samples. Protein levels are indicated by yellow to red colouring, a red shade indicates a higher abundance level. (PDF 7 kb)

Proteins differing significantly in Î yvmB cells from proteome analysis of the cytosolic fraction. A Kruskal-Wallis one-way analysis of variance (ANOVA) was done on the whole proteomic data. (PDF 20 kb)

Multiple sequence alignment of members of the MarR family. The alignment was generated using Multalin (Corpet, 1988, Nucleic Acids Res, 16(22):10881–10890). The aligned proteins are YvmB and OhrR from Bacillus subtilis and MarR from Escherichia coli. Residue numbering is according to entire alignment. Secondary structures elements indicated above the alignment show conservation of a winged helix-turn-helix (wHTH) motif and are based on the E. coli MarR crystal structure, with α-helices represented as open green boxes, β-strands open green arrows and the wing as a filled green box [58]. The HTH domain corresponds to helices α3 and α4, with α4 constituting the recognition helix. The β-hairpin “wing” motif is formed by the β2, W1, β3 structural elements. Helices α1, α5 and α6 form the dimerization domain. (PDF 81 kb)

Proteins differing significantly in Î yvmB cells from proteome analysis of the cytosolic fraction. A Kruskal-Wallis one-way analysis of variance (ANOVA) was done on the whole proteomic data. (PDF 20 kb)

Purification and analysis of YvmBHis-tag protein. (A) SDS-PAGE analysis of purified YvmBHis-tag protein from E. coli cells. (B) Fractionated chromatography on a Superdex 200 h/300 gel filtration column. The YvmBHis-tag protein eluted as a major peak with an apparent molecular mass of 45 ± 5 kDa. (PDF 164 kb)

Promoter regions of the divergent yvnA-yvnB genes and of the yisI gene. (A) The −10 and −35 regions of yvnA (in purple) and the −10 and −35 regions of yvnB (in brown) are indicated according to Nicolas et al. [33]. The predicted YvmB box is indicated in red and a degenerated YvmB box motif is indicated in orange. (B) The −10 and −35 regions of yisI (in brown) is indicated according to Nicolas et al. [33]. The prdicted YvmB box is indicated in red. (PDF 84 kb)

The yvmA-yvmC-cypX locus from B. subtilis and B. licheniformis ATCC 14580. (A) For each gene product of B. subtilis and its equivalent in B. licheniformis ATCC 14580 a percentage of identity is indicated. The position of the genes on each chromosome is given. The yvmC and cypX genes encoding the enzymes involved in pulcherriminic synthesis are represented by purple arrows, the MarR-type regulators YvmB by brown arrows, and the MFS-like transporters YvmA by white arrows. The conserved palindromic motif, palindrome I, is indicated by blue boxes. (B) Alignment of the 42-bp palindromic motifs from the promoter regions of B. subtilis yvmB and B. licheniformis yvmA genes. Stars indicate the seven nucleotides, which are not conserved between the two palindromic sequences. The 14Â bp core region highly conserved in the two sequences is boxed. (PDF 147 kb)

Promoter regions of the divergent yvnA-yvnB genes and of the yisI gene. (A) The −10 and −35 regions of yvnA (in purple) and the −10 and −35 regions of yvnB (in brown) are indicated according to Nicolas et al. [33]. The predicted YvmB box is indicated in red and a degenerated YvmB box motif is indicated in orange. (B) The −10 and −35 regions of yisI (in brown) is indicated according to Nicolas et al. [33]. The prdicted YvmB box is indicated in red. (PDF 84 kb)

Heatmap representation of proteins that were found differentially abundant between the Î yvmB mutant and the wild type strain in the membrane fraction. Heatmap shows the change in protein levels among four independent samples. Protein levels are indicated by yellow to red colouring, a red shade indicates a higher abundance level. (PDF 6 kb)

Proteins differing significantly in Î yvmB cells from proteome analysis of the membrane fraction. A Kruskal-Wallis one-way analysis of variance (ANOVA) was done on the whole proteomic data. (PDF 13 kb)

Effect of iron starvation on the expression of cypX and yvmB. The BFA815 (cypX::lacZ) and BSAS108 (pyvmB’-lacZ) strains were cultivated in LB medium until OD600 of 1. Samples of 2 ml of the cultures were spread onto solid LB medium containing 20 μg.ml−1 X-gal. A drop of 10 μl 10 mM bipyridyl was deposited at the center of each plate. Blue rings corresponded to expression of the fusion in cells around the inhibition zone of bipyridyl drops. (PDF 75 kb)

List of probes used in the gel shift assays. (DOC 32 kb)

List of probes used in the gel shift assays. (DOC 32 kb)