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Cell. 2019 Jul 25;178(3):731-747.e16. doi: 10.1016/j.cell.2019.06.013. Epub 2019 Jun 27.

Deciphering the "m6A Code" via Antibody-Independent Quantitative Profiling.

Author information

1
Department of Molecular Genetics, Weizmann Institute of Science, 7610001 Rehovot, Israel.
2
Medical University of Vienna, Center for Anatomy & Cell Biology, Währinger Straße 13, 1090 Vienna, Austria.
3
Life Sciences Core Facilities, Weizmann Institute of Science, 7610001 Rehovot, Israel.
4
Department of Molecular Genetics, Weizmann Institute of Science, 7610001 Rehovot, Israel. Electronic address: schwartz@weizmann.ac.il.

Abstract

N6-methyladenosine (m6A) is the most abundant modification on mRNA and is implicated in critical roles in development, physiology, and disease. A major limitation has been the inability to quantify m6A stoichiometry and the lack of antibody-independent methodologies for interrogating m6A. Here, we develop MAZTER-seq for systematic quantitative profiling of m6A at single-nucleotide resolution at 16%-25% of expressed sites, building on differential cleavage by an RNase. MAZTER-seq permits validation and de novo discovery of m6A sites, calibration of the performance of antibody-based approaches, and quantitative tracking of m6A dynamics in yeast gametogenesis and mammalian differentiation. We discover that m6A stoichiometry is "hard coded" in cis via a simple and predictable code, accounting for 33%-46% of the variability in methylation levels and allowing accurate prediction of m6A loss and acquisition events across evolution. MAZTER-seq allows quantitative investigation of m6A regulation in subcellular fractions, diverse cell types, and disease states.

PMID:
31257032
DOI:
10.1016/j.cell.2019.06.013

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