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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F21%3A00536199" target="_blank" >RIV/68081707:_____/21:00536199 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216224:14740/21:00118818

Výsledek na webu

<a href="https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/anie.202007184" target="_blank" >https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/anie.202007184</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/anie.202007184" target="_blank" >10.1002/anie.202007184</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

<a href="/cs/project/EF15_003%2F0000477" target="_blank" >EF15_003/0000477: Strukturní gymnastika nukleových kyselin: od molekulárních principů přes biologické funkce k terapeutickým cílům.</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Angewandte Chemie - International Edition

ISSN

1433-7851

e-ISSN

1521-3773

Svazek periodika

60

Číslo periodika v rámci svazku

2

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

10

Strana od-do

865-872

Kód UT WoS článku

000591274200001

EID výsledku v databázi Scopus

2-s2.0-85096655060