Abstract
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.
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Acknowledgments
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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D’Heygère, F., Schwartz, A., Coste, F., Castaing, B., Boudvillain, M. (2015). Monitoring RNA Unwinding by the Transcription Termination Factor Rho from Mycobacterium tuberculosis . In: Boudvillain, M. (eds) RNA Remodeling Proteins. Methods in Molecular Biology, vol 1259. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2214-7_18
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DOI: https://doi.org/10.1007/978-1-4939-2214-7_18
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