A reader-based assay for m6A writers and erasers

TitleA reader-based assay for m6A writers and erasers
Publication TypeJournal Article
Year of Publication2019
AuthorsWiedmer L., Eberle S.A, Bedi R.K, Śledź P., Caflisch A.
JournalAnalytical Chemistry
Volume91
Issue4
Pagination3078-3084
Date Published2019 Feb 19
Type of ArticleResearch Article
ISSN1520-6882
KeywordsDNA methylation, enzyme activity, Enzymes, epitranscriptomics, Fluorescence Resonance Energy Transfer, FRET, High-Throughput Screening Assays, m6A
Abstract

We have developed a homogeneous time-resolved fluorescence (HTRF)-based enzyme assay to measure the catalytic activity of N6-methyladenosine (m6A) methyltransferases and demethylases. The assay detects m6A modifications using the natural m6A-binding proteins (m6A readers). The reaction product or substrate m6A-containing RNA and the m6A reader protein are fluorescently labeled such that their proximity during binding initiates Förster resonance energy transfer (FRET). We show that our HTRF assay can be used for high-throughput screening, which will facilitate the discovery of small-molecule modulators of m6A (de)methylases.

DOI10.1021/acs.analchem.8b05500
pubindex

0244

Alternate JournalAnal. Chem.
PubMed ID30715855
Highlight Role: 
Molecular Interactions