Perturbation of m6A writers reveals two distinct classes of mRNA methylation at internal and 5' sites

Schraga Schwartz, Maxwell R. Mumbach, Marko Jovanovic, Tim Wang, Karolina Maciag, G. Guy Bushkin, Philipp Mertins, Dmitry Ter-Ovanesyan, Naomi Habib, Davide Cacchiarelli, Neville E. Sanjana, Elizaveta Freinkman, Michael E. Pacold, Rahul Satija, Tarjei S. Mikkelsen, Nir Hacohen, Feng Zhang, Steven A. Carr, Eric S. Lander*, Aviv Regev

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

955 Scopus citations

Abstract

N6-methyladenosine (m6A) is a common modification of mRNA with potential roles in fine-tuning the RNA life cycle. Here, we identify a dense network of proteins interacting with METTL3, a component of the methyltransferase complex, and show that three of them (WTAP, METTL14, and KIAA1429) are required for methylation. Monitoring m6A levels upon WTAP depletion allowed the definition of accurate and near single-nucleotide resolution methylation maps and their classification into WTAP-dependent and -independent sites. WTAP-dependent sites are located at internal positions in transcripts, topologically static across a variety of systems we surveyed, and inversely correlated with mRNA stability, consistent with a role in establishing "basal" degradation rates. WTAP-independent sites form at the first transcribed base as part of the cap structure and are present at thousands of sites, forming a previously unappreciated layer of transcriptome complexity. Our data shed light on the proteomic and transcriptional underpinnings of this RNA modification.

Original languageEnglish
Pages (from-to)284-296
Number of pages13
JournalCell Reports
Volume8
Issue number1
DOIs
StatePublished - 10 Jul 2014
Externally publishedYes

Bibliographical note

Funding Information:
This work was supported by NHGRI CEGS P50 HG006193 (A.R.), U54 HG003067 (E.S.L), and Broad Institute Funds. A.R. was supported by an NIH Pioneer Award and HHMI. S.S. was supported by a European Molecular Biology Organization fellowship, and S.S. and D.C. were supported by Human Frontier Science Program fellowships. M.J. was supported by fellowships of the Swiss National Science Foundation for advanced researchers (SNF) and the Marie Sklodowska-Curie IOF.

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