Atomic mutagenesis and SI-labeling of RNA for NMR

The proposed research aims at the establishment of a novel approach the atomic- mutagenesis-and-chemical-shift-fingerprint method to make invisible states of nucleic acids visible. The method will allow to give deep insights into the structure of so called excited states in RNA, which are important for biological function. The research capitalizes on stable isotope labeling of RNA building blocks that carry a carbon/nitrogen replacement in the nucleobase. These building blocks can be positioned within a target nucleic acid at the site where the conformational dynamics occurs. Then, using an NMR spectroscopic method the relaxation dispersion approach - the source of conformational heterogeneity can be pinpointed to a single atom in a nucleic acid and subsequently the function of this atom in processes such as catalysis or protein recognition can be understood.

The proposed research aims at the establishment of a novel approach - the atomic-mutagenesis-and-chemical-shift-fingerprint method - to make invisible states of nucleic acids visible. The method will allow to give deep insights into the structure of so called excited states in RNA, which are important for biological function. The research capitalizes on stable isotope labeling of RNA building blocks that carry a carbon/nitrogen replacement in the nucleobase. These building blocks can be positioned within a target nucleic acid at the site where the conformational dynamics occurs. Then, using an NMR spectroscopic method - the relaxation dispersion approach - the source of conformational heterogeneity can be pinpointed to a single atom in a nucleic acid and subsequently the function of this atom in processes such as catalysis or protein recognition can be understood.