There have been suggestions that Cortisone acetate Rv0560c plays a role related to the biosynthesis of the isoprenoid lipid menaquinone. The fact that this gene is not Mesaconitine expressed during aerobic growth, but upregulated during hypoxia and intraphagosomal growth in macrophages is interesting, indicating that this gene might play a role during infection. The Rv0560c protein is also induced in response to para-aminosalicylic acid, naphthoquinones such as menadione, and plumbagin, as well as the peroxisome proliferator gemfibrozil, and its structural relatives fenofibrate and clofibrate. The range of conditions under which Rv0560c is induced and the huge extent of its induction in response to salicylate are intriguing. The aim of this study was to identify and characterise the promoter of Rv0560c to gain further insight into its expression. Here, we demonstrate for the first time that Rv0560c is expressed from a salicylate-inducible promoter which is highly active. We investigated the induction kinetics of this promoter, which took several days to reach maximal activity and remained highly induced for several weeks. We show that PRv0560c is also induced by structural analogues of salicylate as well as fenofibrates and is mildly induced under conditions of low iron. The 210 and 235 promoter elements, as well as residues involved in its regulation were identified. Our results suggest that this regulatory control is likely to be mediated via a repressor. The data presented here reveal PRv0560c to be a promoter with a high level of induction after salicylate treatment. Salicylate is known to accumulate in iron-depleted mycobacteria as an intermediate during the biosynthesis of the iron-sequestering mycobactin. Thus, as expected the promoter is induced when intracellular levels of iron are depleted. Interestingly induction was not as pronounced as in salicylate-exposed cells, suggesting that the normal intracellular concentrations of salicylate are low. We were interested in the M. tuberculosis gene Rv0560c due to its strikingly high induction in response to salicylate, and its upregulation under conditions mimicking the in vivo environment the bacteria encounter during an infection. Although Rv0560c has been suggested to be in an operon with its upstream gene, our results show that each of the genes does have its own promoter, both of which are active during aerobic growth. Furthermore, we demonstrate that PRv0560c, but not PRv0561c is inducible by salicylate. This is in accordance with a previous transcriptome study showing that Rv0560c, but not Rv0561c is upregulated after salicylate treatment. We found PRv0560c to be a strong promoter as compared to other M. tuberculosis promoters and with slow induction kinetics, taking several days to reach peak activity and remaining stably induced over a course of several weeks. The majority of M. tuberculosis promoters measured using LacZ as a reporter have activity in the range of 100�C1000 MU; examples include the promoters of recA, pknH, embA, mbtB and higBA. There are few examples of promoters with activity in the 10,000 range; for example PrpfA, which is reported as one of the strongest constitutive promoters in M. tuberculosis has an activity of 4500 MU, whereas PwhiB1 has comparable activity to PRv0560c at 800�C15,000 MU. The high level of promoter activity after induction could be due to presence of a perfect match to the canonical 235 element TTGACA, the extended promoter motif, and the high sequence similarity of the 210 element to the consensus sequence TATAAT. Indeed, these attributes are also present in the PwhiB1 promoter with comparable activity.