Moonlighting function of glutamate racemase from Mycobacterium tuberculosis: racemization and DNA gyrase inhibition are two independent activities of the enzyme , by S. Sengupta, S. Ghosh and V. Nagaraja
Microbiology vol. 154, part 9, pp. 2796 - 2803
Fig. S1. Multiple sequence alignment showing highly conserved racemase motifs and the two conserved cysteine residues of M. tuberculosis MurI. The sequences used for the alignment are as follows: 1, Staphylococcus aureus; 2, Bacillus subtilis; 3, Bacillus cereus; 4, Lactobacillus fermenti; 5, Lactobacillus acidophilus; 6, Aquifex sp.; 7, Mycobacterium tuberculosis; 8, Corynebacterium sp.; 9, Clostridium botulinum; 10, Escherichia coli; 11, Pseudomonas aeroginosa. The signature motifs are highlighted by a bold dashed line. The schematic diagram of M. tuberculosis MurI shows the two highly conserved signature motifs along with the putative catalytic cysteine residues. [PDF] (269 kb)
Fig. S2. Expression profile of MurI from pJAM2 constructs in M. smegmatis. M. smegmatis mc2155 cells harbouring either pJAM2 vector or pJAM2-mtmurI constructs (encoding wild-type, WT, or C75SC185S double mutant, DM, forms of MurI) were grown in Middlebrook 7H9 medium at 37°C to mid-exponential phase, induced with 2% acetamide and grown for a further 6 h. The protein expression profiles were checked by 12% SDS-PAGE. Lanes: 1, protein molecular mass markers; 2, cell extract from cells harbouring vector; 3 and 4, uninduced and induced cell extracts, respectively, from transformants harbouring pJAM2-mtmurI (WT); 5 and 6, uninduced and induced cell extracts, respectively, from transformants harbouring pJAM2-mtmurI (DM). [PDF] (637 kb)
Fig. S3. The nucleoid status upon overexpression of MurI resembles that of novobiocin-treated cells. Comparison of nucleoid structures in M. smegmatis mc2155 cells harbouring the pJAM2 vector or pJAM2-mtmurI (WT) or pJAM2-mtmurI (DM) constructs, induced with 2% acetamide. Novobiocin-treated cells were used as positive control showing the relaxed diffused nucleoids for cells upon treatment with this gyrase inhibitor. Left panels, phase-contrast images; middle panels, fluorescent images showing the DAPI-stained nucleoid; right panels, overlay of the two images. Scale bars, 5 μm. [PDF] (436 kb)
Copyright © 2009 Society for General Microbiology.
