NMR Investigation of the Regulation of Gene Expression by RNA Thermometers

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Ribonucleic acid oligonucleotides (RNAs) are involved in the regulation of gene expression. RNAs respond to various external stimuli such as changes in the concentration of small metabolites (riboswitch RNAs) but also changes in temperature (RNA thermometers). The external stimulus is transformed into a change in secondary and tertiary structure, often at allosteric sites. Thereby, gene expression can be regulated both at the level of transcription or translation. In this proposal, we will characterize the structural dynamics of novel RNA thermometers discovered in the Narberhaus group (unpublished results) using static and time-resolved NMR spectroscopy. In the first funding period, we have developed a method that allows us to delineate the stability of individual base pairs based on thermodynamic measurements. In addition, we have developed a NMR temperature jump probe. In the second funding period, we will extend our studies to novel RNA thermometers and will apply time-resolved NMR measurements to determine the kinetics and life times of open signalling states of RNA thermometers.