Isotope labeling for studying RNA by solid-state NMR spectroscopy

Abstract

Nucleic acids play key roles in most biological processes, either in isolation or in complex with proteins. Often they are difficult targets for structural studies, due to their dynamic behavior and high molecular weight. Solid-state nuclear magnetic resonance spectroscopy (ssNMR) provides a unique opportunity to study large biomolecules in a non-crystalline state at atomic resolution. Application of ssNMR to RNA, however, is still at an early stage of development and presents considerable challenges due to broad resonances and poor dispersion. Isotope labeling, either as nucleotide-specific, atom-specific or segmental labeling, can resolve resonance overlaps and reduce the line width, thus allowing ssNMR studies of RNA domains as part of large biomolecules or complexes. In this review we discuss the methods for RNA production and purification as well as numerous approaches for isotope labeling of RNA. Furthermore, we give a few examples that emphasize the instrumental role of isotope labeling and ssNMR for studying RNA as part of large ribonucleoprotein complexes.

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Reproduced with permission from Marchanka et al. (2015)

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Reproduced with permission from Tolbert and Williamson (1996). (© 1996 American Chemical Society. Reproduced with permission from Dayie (2008)

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Acknowledgements

T.C. thanks the DFG for support through Grant CA 294/10 − 1.

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Correspondence to Teresa Carlomagno.

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Marchanka, A., Kreutz, C. & Carlomagno, T. Isotope labeling for studying RNA by solid-state NMR spectroscopy. J Biomol NMR 71, 151–164 (2018). https://doi.org/10.1007/s10858-018-0180-7

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Keywords

  • RNA structure and dynamics
  • Solid-state NMR
  • Chemical synthesis of RNA
  • In vitro RNA transcription
  • Atom selective-labelling of nucleotides