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

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Date

2023-06-07T00:00:00

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Newlands Press Ltd

Abstract

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. � 2023 Future Medicine Ltd.

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Keywords

EMSA, gene expression, MSMEG-5850, Mycobacterium, TetR-like transcription factors, transcriptome analysis

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