Details

Autor(en) / Beteiligte

• Velema, Willem A.
• Park, Hyun Shin
• Kadina, Anastasia
• Orbai, Lucian
• Kool, Eric T.

Titel

Trapping Transient RNA Complexes by Chemically Reversible Acylation

Ist Teil von

• Angewandte Chemie, 2020-12, Vol.132 (49), p.22201-22206

Ort / Verlag

Weinheim: Wiley Subscription Services, Inc

Erscheinungsjahr

2020

Quelle

Wiley Online Library

Beschreibungen/Notizen

• 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. Crosslinking of transient RNA complexes was achieved by using dimeric 2′‐OH acylating reagents. The crosslinks of captured complexes are efficiently reversed by phosphine‐mediated reduction of azide‐trigger groups, thereby enabling convenient downstream analysis.