Organisation and Zur dependent regulation of a MAP specific ABC transporter. (A) Analysis of the mptABC promoter. MAPwt was grown in MB-complete to an OD600 of 1.0 and treated 2 h with 10 μM TPEN. Transcription start sites (TSS) were determined by 5’RACE. Depicted is the putative organisation of the mptABC promoter region [NCBI:NC_002944] (position 4158368 to 4158826). TSS and putative translation start sites (TLS) according to NCBI (NCBI) and 5’RACE results (RACE) are indicated in bold. A putative −10 promoter site is highlighted grey, putative Zur boxes and a ribosome binding site (RBS) are underlined. (B) Heterologous expression and regulation of the mptABC operon in MSMEG. MSMEG was transformed with pMP1102, cultured in MB-complete and treated with TPEN as described above. Gene expression of mptA was analysed by qRT-PCR. Bars represent the relative fold change of the treated transformant (wt+) to the untreated control (wt-) (three independent experiments, mean ± SEM). Statistical analysis was performed using Mann–Whitney test with **p < 0.005. (C) Analysis of Zur binding sites in MAP by FIMO analysis. Upper panel: consensus sequence of Mtb-Zur  used for FIMO. Middle panel: Zur box3 of mptA. Lower panel: mutated mptA Zur box3, black arrows indicate mutated nucleotides. (D) Zur box analysis of the mptABC operon by β-galactosidase assay. MSMEGwt was transformed with the indicated lacZ-reporter plasmids: mptA2, mptA8, mptA3 and mptA2-MUT. Strains were grown in MB-complete and treated with TPEN as described above (black bars) or left untreated (white bars). Proteins were extracted and promoter activity was analysed by β-galactosidase assay (three independent experiments, mean ± SEM). Activity was measured at a wavelength of 405 nm and related to mg protein per ml. Statistical analysis was performed by using the Kruskal-Wallis-Test with *p >0.01 and ***p >0.0001.