Effect of RNA modifications on structure and dynamics probed by NMR

The proposed research project aims to give a deeper understanding of the influence of naturally occurring modifications, such as methylations or formylations, on the RNA folding landscape. To this end stable 13C/15N-isotope labeled modified RNA building blocks will be synthesized, which can be introduced via solid phase synthesis into a target nucleic acid. These novel building blocks in combination with stable isotope labeled standard RNA phosphoramidites will be used in state-of-the-art solution NMR spectroscopic methods to address the modifications influence on structural and dynamic features of nucleic acids. We will incorporate the modified building blocks into a double stranded RNA context and study the influence of the modifications via various biophysical methods, such as NMR and temperature dependent UV spectroscopy. Solution NMR spectroscopy will be used to assess the influence on the folding state by the respective modification in high detail. Structural consequences but also more subtle effects on the dynamic properties will be addressed. Relaxation dispersion NMR methods will be applied to address alterations of the RNA folding landscape. Further rare enolic states earlier found for RNA and DNA wobble base pairs will be investigated in RNA modifications promoting these low populated states. Along this line the stability of base pairs involving modified RNA residues will be determined. The ultimate goal of the research project is to characterize and to understand structural but also dynamic alterations induced by naturally occurring RNA modifications within a double helical RNA arrangement.

The proposed research project aims to give a deeper understanding of the influence of naturally occurring modifications, such as methylations or formylations, on the RNA folding landscape. To this end stable 13C/15N-isotope labeled modified RNA building blocks will be synthesized, which can be introduced via solid phase synthesis into a target nucleic acid. These novel building blocks in combination with stable isotope labeled standard RNA phosphoramidites will be used in state-of-the-art solution NMR spectroscopic methods to address the modification's influence on structural and dynamic features of nucleic acids. We will incorporate the modified building blocks into a double stranded RNA context and study the influence of the modifications via various biophysical methods, such as NMR and temperature dependent UV spectroscopy. Solution NMR spectroscopy will be used to assess the influence on the folding state by the respective modification in high detail. Structural consequences but also more subtle effects on the dynamic properties will be addressed. Relaxation dispersion NMR methods will be applied to address alterations of the RNA folding landscape. Further rare enolic states earlier found for RNA and DNA wobble base pairs will be investigated in RNA modifications promoting these low populated states. Along this line the stability of base pairs involving modified RNA residues will be determined. The ultimate goal of the research project is to characterize and to understand structural but also dynamic alterations induced by naturally occurring RNA modifications within a double helical RNA arrangement.