Mycobacterium tuberculosis transcriptional response to small molecule

We applied RNAseq (Nextseq) technology to study mechanism of action of nitro-containing heterocycle antitubercular JSF-2019, with des-nitro JSF-2026 as control. Briefly, mid-log phase (OD = 0.3) M. tuberculosis culture was treated by 10x MIC of each compound in biological quadruplicates followed by mRNA extraction and RNAseq analysis. We find that JSF-2019 and pretomanid as intracellular NO• donors exhibited distinct transcriptional patterns comparing to extracellular iNOS inducer such as DETA/NO, therefore it suggests distinguished mechanisms of action between intracellular vs. extracellular NO• donors. We also observed that JSF-2019 upregulated a subset of FAS-II genes similar to isoniazid, indicating JSF-2019 inhibits intracellular mycolic acid biosynthesis. Examine M. tuberculosis gene expression at 10x MIC compound treatment for 4 hours.