"End-to-End" stacking of small dsRNA

PELDOR (pulse electron-electron double resonance) is an established method to study intramolecular distances and can give evidence for conformational changes and flexibilities. However, it can also be used to study inter
PELDOR (pulse electron-electron double resonance) is an established method to study intramolecular distances and can give evidence for conformational changes and flexibilities. However, it can also be used to study intermolecular interactions as for example oligerimization. Here, we used PELDOR to study the ‘end-to-end’ stacking of small double stranded (ds)RNAs. For this study, the dsRNA molecules were only singly labelled with the spin label TPA to avoid multi-spin effects and to measure only the intermolecular stacking interactions. It can be shown that small dsRNAs tend to assemble to rod-like structures due to π-π-interactions between the base pairs at the end of the strands. On the one hand, these interactions can influence or complicate measurements aimed at the determining of the structure and dynamics of the dsRNA molecule itself. On the other hand, it can be interesting to study such intermolecular stacking interactions in more detail, as for example their dependence on ion concentration. We quantitatively determined the stacking probability as a function of the monovalent NaCl salt and the dsRNA concentration. From this data the dissociation constant Kd was deduced and found to depend on the ratio between the NaCl salt and dsRNA concentrations. Additionally, the distances and distance distributions obtained predict a model for the stacking geometry of dsRNAs. Introducing a nucleotide overhangs at one end of the dsRNA molecule restricts the stacking to the other end, leading only to dimer formations. Introducing such an overhang at both ends of the dsRNA molecule fully suppresses stacking, as we could demonstrate by PELDOR experiments quantitatively.
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Metadaten
Author:Nicole Erlenbach, Christian Grünewald, Bisera Krstic, Alexander Heckel, Thomas F. Prisner
URN:urn:nbn:de:hebis:30:3-512662
DOI:http://dx.doi.org/10.1261/rna.068130.118
ISSN:1469-9001
ISSN:1355-8382
Pubmed Id:http://www.ncbi.nlm.nih.gov/pubmed?term=30404925
Parent Title (English):RNA
Publisher:Laboratory Press ; HighWire Press
Place of publication:Cold Spring Harbor, NY ; Stanford, Calif.
Document Type:Article
Language:English
Year of Completion:2018
Date of first Publication:2018/11/07
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2020/01/20
Tag:EPR; PELDOR/DEER; dsRNA; stacking
Volume:25
Issue:2
Pagenumber:26
First Page:239
Last Page:246
Note:
This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
HeBIS PPN:459856219
Institutes:Biochemie und Chemie
Starker Start ins Studium: Qualitätspakt Lehre
Zentrum für Biomolekulare Magnetische Resonanz (BMRZ)
Dewey Decimal Classification:540 Chemie und zugeordnete Wissenschaften
570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Licence (English):License LogoCreative Commons - Namensnennung-Nicht kommerziell 4.0

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