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MSMEG_5850, a stress-induced TetR protein, involved in global transcription regulation in Mycobacterium smegmatis

    Parul Singh

    Department of Biotechnology, BMS Block-1, Sector-25, Panjab University, Chandigarh, 160014, India

    ,
    Arbind Kumar

    Department of Biotechnology, BMS Block-1, Sector-25, Panjab University, Chandigarh, 160014, India

    Current Address: Fellow Scientist, CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, 176061, India

    ,
    Ravindresh Chhabra

    Department of Biochemistry, Central University of Punjab, Bathinda, 151001, India

    ,
    Kashmir Singh

    Department of Biotechnology, BMS Block-1, Sector-25, Panjab University, Chandigarh, 160014, India

    &
    Jagdeep Kaur

    *Author for correspondence: Tel.: +91 0172 253 4085;

    E-mail Address: jagsekhon@pu.ac.in

    Department of Biotechnology, BMS Block-1, Sector-25, Panjab University, Chandigarh, 160014, India

    Published Online:https://doi.org/10.2217/fmb-2022-0238

    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.

    Graphical abstract

    Papers of special note have been highlighted as: • of interest; •• of considerable interest

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