Transcriptional pausing at the translation start site operates as a critical checkpoint for riboswitch regulation

Adrien Chauvier, Frédéric Picard-Jean, Jean-Christophe Berger-Dancause, Laurène Bastet, Mohammad Reza Naghdi, Audrey Dubé, Pierre Turcotte, Jonathan Perreault and Daniel A. Lafontaine ()
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Adrien Chauvier: Faculty of Science, RNA Group, Université de Sherbrooke
Frédéric Picard-Jean: Faculty of Science, RNA Group, Université de Sherbrooke
Jean-Christophe Berger-Dancause: Faculty of Science, RNA Group, Université de Sherbrooke
Laurène Bastet: Faculty of Science, RNA Group, Université de Sherbrooke
Mohammad Reza Naghdi: INRS-Institut Armand-Frappier
Audrey Dubé: Faculty of Science, RNA Group, Université de Sherbrooke
Pierre Turcotte: Faculty of Science, RNA Group, Université de Sherbrooke
Jonathan Perreault: INRS-Institut Armand-Frappier
Daniel A. Lafontaine: Faculty of Science, RNA Group, Université de Sherbrooke

Nature Communications, 2017, vol. 8, issue 1, 1-12

Abstract: Abstract On the basis of nascent transcript sequencing, it has been postulated but never demonstrated that transcriptional pausing at translation start sites is important for gene regulation. Here we show that the Escherichia coli thiamin pyrophosphate (TPP) thiC riboswitch contains a regulatory pause site in the translation initiation region that acts as a checkpoint for thiC expression. By biochemically probing nascent transcription complexes halted at defined positions, we find a narrow transcriptional window for metabolite binding, in which the downstream boundary is delimited by the checkpoint. We show that transcription complexes at the regulatory pause site favour the formation of a riboswitch intramolecular lock that strongly prevents TPP binding. In contrast, cotranscriptional metabolite binding increases RNA polymerase pausing and induces Rho-dependent transcription termination at the checkpoint. Early transcriptional pausing may provide a general mechanism, whereby transient transcriptional windows directly coordinate the sensing of environmental cues and bacterial mRNA regulation.

Date: 2017
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DOI: 10.1038/ncomms13892

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