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Trapping Transient RNA Complexes by Chemically Reversible Acylation.

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.

SUBMITTER: Velema WA

PROVIDER: S-EPMC7988885 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Trapping Transient RNA Complexes by Chemically Reversible Acylation.

Velema Willem A WA Park Hyun Shin HS Kadina Anastasia A Orbai Lucian L Kool Eric T ET

Angewandte Chemie (International ed. in English) 20200928 49

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 ...[more]

PMID: 32845055

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