TY  - JOUR
A1  - Broft, Patrizia
A1  - Dzatko, Simon
A1  - Krafcikova, Michaela
A1  - Wacker, Anna
A1  - Hänsel-Hertsch, Robert
A1  - Dötsch, Volker
A1  - Trantirek, Lukáš
A1  - Schwalbe, Harald
T1  - In-cell NMR spectroscopy of functional Riboswitch aptamers in eukaryotic cells
T2  - Angewandte Chemie
N2  - We report here the in-cell NMR-spectroscopic observation of the binding of the cognate ligand 2′-deoxyguanosine to the aptamer domain of the bacterial 2′-deoxyguanosine-sensing riboswitch in eukaryotic cells, namely Xenopus laevis oocytes and in human HeLa cells. The riboswitch is sufficiently stable in both cell types to allow for detection of binding of the ligand to the riboswitch. Most importantly, we show that the binding mode established by in vitro characterization of this prokaryotic riboswitch is maintained in eukaryotic cellular environment. Our data also bring important methodological insights: Thus far, in-cell NMR studies on RNA in mammalian cells have been limited to investigations of short (<15 nt) RNA fragments that were extensively modified by protecting groups to limit their degradation in the intracellular space. Here, we show that the in-cell NMR setup can be adjusted for characterization of much larger (≈70 nt) functional and chemically non-modified RNA.
KW  - aptamers
KW  - 2'-deoxyguanosine riboswitch
KW  - HeLa cells
KW  - RNA structures
KW  - structural biology
Y1  - 2020
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/63856
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-638561
SN  - 1521-3773
N1  - Open access funding enabled and organized by Projekt DEAL.
VL  - 60
IS  - 2
SP  - 865
EP  - 872
PB  - Wiley-VCH
CY  - Weinheim
ER  -