Monitoring RNA Unwinding by the Transcription Termination Factor Rho from Mycobacterium tuberculosis
Transcription termination factor Rho is a ring-shaped, homo-hexamieric RNA translocase that dissociates transcription elongation complexes and transcriptional RNA–DNA duplexes (R-loops) in bacteria. The molecular mechanisms underlying these biological functions have been essentially studied with Rho enzymes from Escherichia coli or close Gram-negative relatives. However, phylo-divergent Rho factors may have distinct properties. Here, we describe methods for the preparation and in vitro characterization (ATPase and helicase activities) of the Rho factor from Mycobacterium tuberculosis, a specimen with uncharacteristic molecular and enzymatic features. These methods set the stage for future studies aimed at better defining the diversity of enzymatic properties of Rho across the bacterial kingdom.
Key wordsRho Transcription Termination Ring shaped Helicase Hexamer RNA Molecular motor
This work was supported by a joint Ph.D. scholarship from the University of Orléans and Région Centre to F.d.H and by CNRS core funding to M.B.
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