TY - JOUR

T1 - Isotope labeling for studying RNA by solid-state NMR spectroscopy

AU - Marchanka, Alexander

AU - Kreutz, Christoph

AU - Carlomagno, Teresa

N1 - © 2018, Springer Science Business Media B.V., part of Springer Nature

PY - 2018/7

Y1 - 2018/7

N2 - 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.

AB - 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.

KW - Atom selective-labelling of nucleotides

KW - Chemical synthesis of RNA

KW - In vitro RNA transcription

KW - RNA structure and dynamics

KW - Solid-state NMR

UR - http://www.scopus.com/inward/record.url?scp=85045289123&partnerID=8YFLogxK

U2 - 10.1007/s10858-018-0180-7

DO - 10.1007/s10858-018-0180-7

M3 - Article

C2 - 29651587

AN - SCOPUS:85045289123

VL - 71

SP - 151

EP - 164

JO - Journal of Biomolecular NMR

JF - Journal of Biomolecular NMR

SN - 0925-2738

IS - 3

ER -