1 (Lighthouse data). The annotation of S. aureus N315 was used for protein identification and denotation. Peptide mixtures that yielded at least twice a Mowes score of at least 50 and a sequence coverage

of at least 30% were regarded as positive identifications. Proteins that failed to exceed the 30% sequence coverage cut-off were subjected to MALDI-MS/MS [73]. Database searches were performed using the Mascot search engine with the protein databases of S. aureus strain N315. Protein quantitation approaches The 2D gel image analysis was performed with the software “”Delta2D”" (DECODON GmbH, Greifswald, Germany). Three different data sets were analyzed in order to screen for differences in the amount of cytoplasmic proteins identified Selleckchem Everolimus on 2D gels. Detection of glucose, acetate and lactate EPZ015666 mw The concentrations of glucose, acetate and lactate in the supernatants were determined using commercially available

kits (Boehringer) according to the manufacturer’s instructions. Urease assay McFarland 0.5-standard cell suspensions were diluted 100-fold in urea medium [74] and incubated in 12-well plates at 37° for 24 hours. In parallel, colony forming units (cfu/ml) were determined. Acknowledgements This study was supported by the Swiss National Science Foundation grants 310000-117707 (to BBB), 3100A0-112370/1 (to JS), and 3100A0-116075/1 (to PF) and the Deutsche Forschungsgemeinschaft (grant Bi 1350/1-1 to MB). Electronic supplementary material Additional file 1: Genes with lower expression in wild-type versus Δ ccpA mutant. The table represents genes

showing a lower gene expression in the from wild-type than the ΔccpA mutant (wt/mutant ratio ≤ 0.5). Cells were grown in LB, without glucose addition. (DOC 236 KB) Additional file 2: Genes with higher expression in wild-type versus Δ ccpA mutant. The table represents genes showing a higher gene expression in the wild-type than the ΔccpA mutant (wt/mutant ratio ≥ 2.0). Cells were grown in LB, without glucose addition. (DOC 210 KB) Additional file 3: CcpA-dependent down-regulation by glucose. The table shows genes found to be subject to down-regulation by glucose in a CcpA-dependent manner (with/without glucose ratio of 0.5 or lower in wild-type, with/without glucose ratio of approximately 1, but below 2 in the mutant). (DOC 284 KB) Additional file 4: CcpA-dependent up-regulation by glucose. The table shows genes found to be subject to up-regulation by glucose in a CcpA-dependent manner (with/without glucose ratio of 2 or higher in wild-type, with/without glucose ratio of approximately 1, but below 2 in the mutant). (DOC 258 KB) Additional file 5: Primers used for the construction of DIG-labelled DNA probes. (DOC 36 KB) References 1. Fujita Y: Carbon catabolite control of the metabolic network in Bacillus subtilis. Bioscience, Biotechnology, and Biochemistry 2009,73(2):245–259.CrossRefPubMed 2.