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Trapping Transient RNA Complexes by Chemically Reversible Acylation

Trapping Transient RNA Complexes by Chemically Reversible Acylation RNA‐RNA interactions are essential for biology, but they can be difficult to study due to their transient nature. While crosslinking strategies can in principle be used to trap such interactions, virtually all existing strategies for crosslinking are poorly reversible, chemically modifying the RNA and hindering molecular analysis. We describe a soluble crosslinker design (BINARI) that reacts with RNA through acylation. We show that it efficiently crosslinks noncovalent RNA complexes with mimimal sequence bias and establish that the crosslink can be reversed by phosphine reduction of azide trigger groups, thereby liberating the individual RNA components for further analysis. The utility of the new approach is demonstrated by reversible protection against nuclease degradation and trapping transient RNA complexes of E. coli DsrA‐rpoS derived bulge‐loop interactions, which underlines the potential of BINARI crosslinkers to probe RNA regulatory networks. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Angewandte Chemie International Edition Wiley

Trapping Transient RNA Complexes by Chemically Reversible Acylation

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References (56)

Publisher
Wiley
Copyright
© 2020 Wiley‐VCH GmbH
ISSN
1433-7851
eISSN
1521-3773
DOI
10.1002/anie.202010861
Publisher site
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Abstract

RNA‐RNA interactions are essential for biology, but they can be difficult to study due to their transient nature. While crosslinking strategies can in principle be used to trap such interactions, virtually all existing strategies for crosslinking are poorly reversible, chemically modifying the RNA and hindering molecular analysis. We describe a soluble crosslinker design (BINARI) that reacts with RNA through acylation. We show that it efficiently crosslinks noncovalent RNA complexes with mimimal sequence bias and establish that the crosslink can be reversed by phosphine reduction of azide trigger groups, thereby liberating the individual RNA components for further analysis. The utility of the new approach is demonstrated by reversible protection against nuclease degradation and trapping transient RNA complexes of E. coli DsrA‐rpoS derived bulge‐loop interactions, which underlines the potential of BINARI crosslinkers to probe RNA regulatory networks.

Journal

Angewandte Chemie International Edition – Wiley

Published: Dec 1, 2020

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