Trapping Transient RNA Complexes by Chemically Reversible Acylation.

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

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

Afiliación

Velema WA; Institute for Molecules and Materials, Radboud University, Nijmegen, 6525, AJ, The Netherlands.
Park HS; Department of Chemistry, Stanford University, Stanford, CA, 94305, USA.
Kadina A; Department of Chemistry, Stanford University, Stanford, CA, 94305, USA.
Orbai L; Cell Data Sciences, 46127 Landing Pkwy, Fremont, CA, 94538, USA.
Kool ET; Department of Chemistry, Stanford University, Stanford, CA, 94305, USA.

Angew Chem Int Ed Engl ; 59(49): 22017-22022, 2020 12 01.

Article en En | MEDLINE | ID: mdl-32845055

RESUMEN

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.

Asunto(s)

Azidas/metabolismo; Reactivos de Enlaces Cruzados/metabolismo; Escherichia coli/química; Fosfinas/metabolismo; ARN Bacteriano/metabolismo; Acilación; Azidas/química; Reactivos de Enlaces Cruzados/química; Escherichia coli/metabolismo; Fosfinas/química; ARN Bacteriano/química

Palabras clave

RNA complexes; acylation; bioorthogonal chemistry; crosslinking; nucleic acids

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Fosfinas / Azidas / ARN Bacteriano / Reactivos de Enlaces Cruzados / Escherichia coli Idioma: En Revista: Angew Chem Int Ed Engl Año: 2020 Tipo del documento: Article País de afiliación: Países Bajos Pais de publicación: Alemania

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