RNA-sequencing of Mtb H37Rv, Mtb?iscS, iscS complemented, Mtb-sufS knockdown, ?iscS-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. Overall design: This study involves determining the mRNA transcriptome profiles for the strains Mtb H37Rv, Mtb?iscS, iscS complemented, Mtb-sufS knockdown, ?iscS-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.