Probing RNA Structure
            <i>In Vitro</i>
            with Enzymes and Chemicals

Helfer, Anne‐Catherine; Romilly, Cédric; Chevalier, Clément; Lioliou, Efthimia; Marzi, Stefano; Romby, Pascale

doi:10.1002/9783527647064.ch10

Cited by 2 publications

(1 citation statement)

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“…The reactions were stopped by precipitation of the RNA by adding 20 µL of a solution containing 1M guanidine thiocyanate, 0.167% N-lauryl sarcosine, 10 mM DTT and 83% isopropanol. The enzymatic cleavages were detected by primer extension using an end-labeled oligonucleotide (CC1537) and reverse transcriptase as previously described ( Helfer et al, 2014 ). The cleavage sites were analyzed after fractionation of the DNA fragments on a 10% polyacrylamide-8 M urea gel-TBE 1X followed by autoradiography.…”

Section: Methodsmentioning

confidence: 99%

sRNA-mediated activation of gene expression by inhibition of 5'-3’ exonucleolytic mRNA degradation

Durand

Braun

Helfer

et al. 2017

eLife

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Post-transcriptional control by small regulatory RNA (sRNA) is critical for rapid adaptive processes. sRNAs can directly modulate mRNA degradation in Proteobacteria without interfering with translation. However, Firmicutes have a fundamentally different set of ribonucleases for mRNA degradation and whether sRNAs can regulate the activity of these enzymes is an open question. We show that Bacillus subtilis RoxS, a major trans-acting sRNA shared with Staphylococus aureus, prevents degradation of the yflS mRNA, encoding a malate transporter. In the presence of malate, RoxS transiently escapes from repression by the NADH-sensitive transcription factor Rex and binds to the extreme 5’-end of yflS mRNA. This impairs the 5’-3’ exoribonuclease activity of RNase J1, increasing the half-life of the primary transcript and concomitantly enhancing ribosome binding to increase expression of the transporter. Globally, the different targets regulated by RoxS suggest that it helps readjust the cellular NAD+/NADH balance when perturbed by different stimuli.DOI: http://dx.doi.org/10.7554/eLife.23602.001

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Section: Methodsmentioning

confidence: 99%