Universität zu Köln

Broft, P., Dzatko, S., Krafcikova, M., Wacker, A., Haensel-Hertsch, Robert, Dotsch, Volker, Trantirek, L. and Schwalbe, Harald . In-Cell NMR Spectroscopy of Functional Riboswitch Aptamers in Eukaryotic Cells. Angew. Chem.-Int. Edit.. WEINHEIM: WILEY-V C H VERLAG GMBH. ISSN 1521-3773

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Abstract

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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Broft, P. UNSPECIFIED UNSPECIFIED UNSPECIFIED Dzatko, S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Krafcikova, M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Wacker, A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Haensel-Hertsch, Robert UNSPECIFIED UNSPECIFIED UNSPECIFIED Dotsch, Volker UNSPECIFIED UNSPECIFIED UNSPECIFIED Trantirek, L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Schwalbe, Harald UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-311838
DOI:	10.1002/anie.202007184
Journal or Publication Title:	Angew. Chem.-Int. Edit.
Publisher:	WILEY-V C H VERLAG GMBH
Place of Publication:	WEINHEIM
ISSN:	1521-3773
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language G-QUADRUPLEX STRUCTURES; SENSING RIBOSWITCH; LIGAND-BINDING; STRUCTURAL BASIS; RNA; DNA; PROTEINS; SIGNALS; VITRO Multiple languages Chemistry, Multidisciplinary Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/31183