NMR-VitAmin

In-cell Nuclear Magnetic Resonance spectroscopy (in-cell NMR) represents a novel biophysical tool to investigate structural transformations of biomolecules at the atomic level inside living cells. Indeed, NMR in vivo and/or ex vivo has recently undergone great advances, and remarkable results have been obtained by monitoring incorporation of post-translational modifications in proteins within a cellular context. These studies demonstrated that in-extract and in-cell NMR are particularly well suited to monitor the maturation process of biological macromolecules, such as proteins. Along the same lines, our project aims at deciphering RNA, mostly transfer RNA (tRNA) maturation events with in-cell NMR. The genomic and the improvements of analytical tools have led to a renaissance of the tRNA modification field. Some human diseases have now been clearly linked to the absence of tRNA modifications. The role of tRNA modifications as regulatory mechanisms needs to be studied further. In addition, many of the in vivo roles of tRNA modifications remain undisclosed. This proposal aims at deciphering the maturation process of tRNA at an atomic level, including information on the chronology of the events leading to the biosynthesis of a fully mature and functional tRNA. The originality and the challenge of this proposal entitled “NMR View of tRNA maturation” (NMR-VitAmin) resides in the use of in-extracts and/or in-cell NMR to elucidate some critical aspects of tRNA biosynthesis and maturation under the conditions of a living cell. NMR will be also used as a screening tool in-extracts to evaluate the action of an RNA-binding chemical library on the tRNA maturation process. Actually, we also aim at interfering with this highly regulated process, in which a misbalance in the level of tRNA modifications is often associated with pathologies.