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:

Impact factor

Impact factor: 16.823

RIV identification code

RIV/00216224:14740/21:00118818

Organization unit

Central European Institute of Technology

UT WoS

000591274200001

Keywords in English

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

Tags

International impact, Reviewed
Změněno: 4/3/2022 13:34, Mgr. Pavla Foltynová, Ph.D.

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
LM2015062, research and development project
Name: Národní infrastruktura pro biologické a medicínské zobrazování
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)