In-Cell NMR Spectroscopy of Functional Riboswitch Aptamers in
Eukaryotic Cells

J 2021

In-Cell NMR Spectroscopy of Functional Riboswitch Aptamers in Eukaryotic Cells

BROFT, P., Šimon DŽATKO, Michaela KRAFČÍKOVÁ, A. WACKER, Robert HANSEL-HERTSCH et. al.

Basic information

Original name

In-Cell NMR Spectroscopy of Functional Riboswitch Aptamers in Eukaryotic Cells

Authors

BROFT, P., Šimon DŽATKO (703 Slovakia, belonging to the institution), Michaela KRAFČÍKOVÁ (703 Slovakia, belonging to the institution), A. WACKER, Robert HANSEL-HERTSCH, Volker DOTSCH, Lukáš TRANTÍREK (203 Czech Republic, guarantor, belonging to the institution) and Harald SCHWALBE

Edition

Angewandte Chemie International Edition, Weinheim, Wiley-VCH Verlag, 2021, 1433-7851

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 16.823

RIV identification code

RIV/00216224:14740/21:00118818

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1002/anie.202007184

UT WoS

000591274200001

Keywords in English

aptamers; 2' -deoxyguanosine riboswitch; HeLa cells; RNA structures; structural biology

Abstract

V originále

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 (approximate to 70 nt) functional and chemically non-modified RNA.

Links

GX19-26041X, research and development project

Name: Strukturní biologie nové generace: Od izolovaných molekul k buňkám, od buněk ke tkáním.

Investor: Czech Science Foundation

LM2015043, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

LM2015064, research and development project

Name: Český národní uzel Evropské infrastruktury pro translační medicínu (Acronym: EATRIS-ERIC-CZ)

Investor: Ministry of Education, Youth and Sports of the CR

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

871037, interní kód MU

Name: iNEXT-Discovery: Infrastructure for transnational access and discovery in integrated structural biology (Acronym: iNEXT- Discovery)

Investor: European Union, RI Research Infrastructures (Excellent Science)

90062, large research infrastructures

Name: Czech-BioImaging

Citovat

BROFT, P., Šimon DŽATKO, Michaela KRAFČÍKOVÁ, A. WACKER, Robert HANSEL-HERTSCH, Volker DOTSCH, Lukáš TRANTÍREK and Harald SCHWALBE. In-Cell NMR Spectroscopy of Functional Riboswitch Aptamers in Eukaryotic Cells. Angewandte Chemie International Edition. Weinheim: Wiley-VCH Verlag, 2021, vol. 60, No 2, p. 865-872. ISSN 1433-7851. Available from: https://dx.doi.org/10.1002/anie.202007184.

@article{1726480,
   author = {Broft, P. and Džatko, Šimon and Krafčíková, Michaela and Wacker, A. and HanselandHertsch, Robert and Dotsch, Volker and Trantírek, Lukáš and Schwalbe, Harald},
   article_location = {Weinheim},
   article_number = {2},
   doi = {http://dx.doi.org/10.1002/anie.202007184},
   keywords = {aptamers; 2' -deoxyguanosine riboswitch; HeLa cells; RNA structures; structural biology},
   language = {eng},
   issn = {1433-7851},
   journal = {Angewandte Chemie International Edition},
   title = {In-Cell NMR Spectroscopy of Functional Riboswitch Aptamers in Eukaryotic Cells},
   url = {https://doi.org/10.1002/anie.202007184},
   volume = {60},
   year = {2021}
}

TY  - JOUR
ID  - 1726480
AU  - Broft, P. - Džatko, Šimon - Krafčíková, Michaela - Wacker, A. - Hansel-Hertsch, Robert - Dotsch, Volker - Trantírek, Lukáš - Schwalbe, Harald
PY  - 2021
TI  - In-Cell NMR Spectroscopy of Functional Riboswitch Aptamers in Eukaryotic Cells
JF  - Angewandte Chemie International Edition
VL  - 60
IS  - 2
SP  - 865-872
EP  - 865-872
PB  - Wiley-VCH Verlag
SN  - 14337851
KW  - aptamers
KW  - 2' -deoxyguanosine riboswitch
KW  - HeLa cells
KW  - RNA structures
KW  - structural biology
UR  - https://doi.org/10.1002/anie.202007184
L2  - https://doi.org/10.1002/anie.202007184
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 (approximate to 70 nt) functional and chemically non-modified RNA.
ER  -

BROFT, P., Šimon DŽATKO, Michaela KRAFČÍKOVÁ, A. WACKER, Robert HANSEL-HERTSCH, Volker DOTSCH, Lukáš TRANTÍREK and Harald SCHWALBE. In-Cell NMR Spectroscopy of Functional Riboswitch Aptamers in Eukaryotic Cells. \textit{Angewandte Chemie International Edition}. Weinheim: Wiley-VCH Verlag, 2021, vol.~60, No~2, p.~865-872. ISSN~1433-7851. Available from: https://dx.doi.org/10.1002/anie.202007184.

Detailed Information on Publication Record