The Mycobacterium tuberculosis (Mtb) DosRST two-component regulatory system promotes the survival of Mtb during non-replicating persistence (NRP). NRP bacteria help drive the long course of tuberculosis therapy; therefore, chemical inhibition of DosRST may inhibit the ability of Mtb to establish persistence and thus shorten treatment. Using a DosRST-dependent fluorescent Mtb reporter strain, a whole-cell phenotypic high-throughput screen of a ~540,000 compound small-molecule library was conducted. The screen discovered novel inhibitors of the DosRST regulon, including three compounds that were subject to follow-up studies: artemisinin, HC102A and HC103A. Under hypoxia, all three compounds inhibit Mtb-persistence-associated physiological processes, including triacylglycerol synthesis, survival and antibiotic tolerance. Artemisinin functions by disabling the heme-based DosS and DosT sensor kinases by oxidizing ferrous heme and generating heme–artemisinin adducts. In contrast, HC103A inhibits DosS and DosT autophosphorylation activity without targeting the sensor kinase heme.
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- Supplementary Text and Figures (8,456 KB)
Supplementary Results, Supplementary Tables 1 and 2 and Supplementary Figures 1–8
- Supplementary Note (160 KB)
- Supplementary Dataset 1 (169 KB)
Differential gene expression data of WT Mtb treated with inhibitors and the DMSO treated DosR mutant.
- Supplementary Dataset 2 (83 KB)
Differential gene expression data of the DosR mutant treated with the inhibitors.
- Supplementary Dataset 3 (3,400 KB)
Complete gene expression tables for transcriptional profiling experiments.