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Electrophilic activity-based RNA probes reveal a self-alkylating RNA for RNA labeling.

ABSTRACT: Probes that form covalent bonds with RNA molecules on the basis of their chemical reactivity would advance our ability to study the transcriptome. We developed a set of electrophilic activity-based RNA probes designed to react with unusually nucleophilic RNAs. We used these probes to identify reactive genome-encoded RNAs, resulting in the discovery of a 42-nt catalytic RNA from an archaebacterium that reacts with a 2,3-disubstituted epoxide at N7 of a specific guanosine. Detailed characterization of the catalytic RNA revealed the structural requirements for reactivity. We developed this catalytic RNA into a general tool to selectively conjugate a small molecule to an RNA of interest. This strategy enabled up to 500-fold enrichment of target RNA from total mammalian RNA or from cell lysate. We demonstrated the utility of this approach by selectively capturing proteins in yeast cell lysate that bind the ASH1 mRNA.

SUBMITTER: McDonald RI

PROVIDER: S-EPMC4232462 | biostudies-literature | 2014 Dec

REPOSITORIES: biostudies-literature

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Electrophilic activity-based RNA probes reveal a self-alkylating RNA for RNA labeling.

McDonald Richard I RI Guilinger John P JP Mukherji Shankar S Curtis Edward A EA Lee Won I WI Liu David R DR

Nature chemical biology 20141012 12

Probes that form covalent bonds with RNA molecules on the basis of their chemical reactivity would advance our ability to study the transcriptome. We developed a set of electrophilic activity-based RNA probes designed to react with unusually nucleophilic RNAs. We used these probes to identify reactive genome-encoded RNAs, resulting in the discovery of a 42-nt catalytic RNA from an archaebacterium that reacts with a 2,3-disubstituted epoxide at N7 of a specific guanosine. Detailed characterizatio ...[more]

PMID: 25306441

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