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Angew Chem Int Ed Engl. 2018 May 22;57(21):6342-6346. doi: 10.1002/anie.201800188. Epub 2018 Mar 22.

Enzymatic or In Vivo Installation of Propargyl Groups in Combination with Click Chemistry for the Enrichment and Detection of Methyltransferase Target Sites in RNA.

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Institute of Biochemistry, Department of Chemistry, University of Münster, Wilhelm-Klemm-Straße 2, 48149, Münster, Germany.
Max Planck Research Group for RNA Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstraße 20, 48149, Münster, Germany.


m6 A is the most abundant internal modification in eukaryotic mRNA. It is introduced by METTL3-METTL14 and tunes mRNA metabolism, impacting cell differentiation and development. Precise transcriptome-wide assignment of m6 A sites is of utmost importance. However, m6 A does not interfere with Watson-Crick base pairing, making polymerase-based detection challenging. We developed a chemical biology approach for the precise mapping of methyltransferase (MTase) target sites based on the introduction of a bioorthogonal propargyl group in vitro and in cells. We show that propargyl groups can be introduced enzymatically by wild-type METTL3-METTL14. Reverse transcription terminated up to 65 % at m6 A sites after bioconjugation and purification, hence enabling detection of METTL3-METTL14 target sites by next generation sequencing. Importantly, we implemented metabolic propargyl labeling of RNA MTase target sites in vivo based on propargyl-l-selenohomocysteine and validated different types of known rRNA methylation sites.


N6-methyladenosine; RNA; metabolic labeling; ribose methylation; transferases


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