MSMEG 5850, a stress-induced TetR protein, involved in global transcription regulation in Mycobacterium smegmatis - Supplementary Figure 2

Aim: To decipher the role of MSMEG 5850 in the physiology of mycobacteria. Methods: MSMEG 5850 was knocked out and RNA sequencing was performed. MSMEG 5850 protein was purified from the Escherichia coli pET28a system. Electrophoretic mobility shift assay and size exclusion chromatography were used to determine the binding of MSMEG 5850 to its motif and binding stoichiometry. The effect of nutritional stress was monitored. Results: Transcriptome analysis revealed the differential expression of 148 genes in an MSMEG 5850 knockout strain. MSMEG 5850 had control over 50 genes because those genes had a binding motif upstream of their sequence. The electrophoretic mobility shift assay showed MSMEG 5850 bound to its motif as a monomer. MSMEG 5850 was upregulated under nutritional stress and promoted the survival of mycobacteria. Conclusion: The study confirms the role of MSMEG 5850 in global transcriptional regulation.