Synthesis of [7-15N]-GTPs for RNA structure and dynamics by NMR spectroscopy

Kehinde M Taiwo,T Kwaku Dayie,Christoph Kreutz,Stefan Hilber,Hyeyeon Nam,Lukasz T Olenginski,Felix Nußbaumer

doi:10.1007/s00706-022-02892-1

Abstract

Several isotope-labeling strategies have been developed for the study of RNA by nuclear magnetic resonance (NMR) spectroscopy. Here, we report a combined chemical and enzymatic synthesis of [7-15N]-guanosine-5′-triphosphates for incorporation into RNA via T7 RNA polymerase-based in vitro transcription. We showcase the utility of these labels to probe both structure and dynamics in two biologically important RNAs.Graphical abstract

Highlights

RNAs, once thought of as an intermediate in the flow of genetic information from DNA to proteins, are credited with playing a central role in many cellular functions [1,2,3,4]
A necessary nuclear magnetic resonance (NMR) parameter for structure determination is proton–proton distances, which is provided by nuclear Overhauser effect spectroscopy (NOESY) experiments
We report the synthesis of atom- labeled [7-15N]-GTPs for use in in vitro transcription to make RNA for NMR analysis

Summary

Introduction

RNAs, once thought of as an intermediate in the flow of genetic information from DNA to proteins, are credited with playing a central role in many cellular functions [1,2,3,4]. RNAs have increasingly become the target of structural and therapeutic efforts [5,6,7]. Among the different techniques available to study RNA, nuclear magnetic

Present Address

Results and discussion

Conclusion