RNA-sequencing of Mtb H37Rv, MtbΔiscS, iscS complemented, Mtb-sufS knockdown, ΔiscS-sufS knockdown

Iron-sulfur (Fe-S) cluster containing proteins are a subset of proteins with crucial functions in the maintenance of cellular physiology throughout all kingdoms of life. The systems involved in the biogenesis and repair of Fe-S clusters hence plays important role in fine-tuning the availability and functionality of Fe-S proteins. Two of the systems known in bacteria are, Isc and Suf. Compared to the facultative anaerobe, E. coli, which codes for the two multi-genic Fe-S biogenesis systems; Mtb Fe-S biogenesis machinery is skewed with a multi-genic Suf system (sufRBDCSUT) and a single gene of Isc system (iscS). Several Fe-S proteins are deployed by Mtb to maintain cellular homeostasis and survival in a hostile host environment. Hence, we determine the transcriptome of Mtb on depletion of the two key enzymes of Fe-S biogenesis- IscS and sufS, that could help understand the role and regulation between the two systems in the human pathogen Mtb. This study involves determining the mRNA transcriptome profiles for the strains Mtb H37Rv, MtbΔiscS, iscS complemented, Mtb-sufS knockdown, ΔiscS-sufS knockdown; grown under standard in vitro conditions in 7H9+ADS+Tween80 at 37°C and 180 rpm, till cultures reached ~0.4 OD600. Knockdown was obtained by addition of 200 ng/ml anhydrotetracycline (ATc). Three independent experiments were done for each of the strains.