Interactions, localization, and phosphorylation of the m6A generating METTL3–METTL14–WTAP complex
Open accessAbstract
N 6 -methyladenine (m 6 A) is found on many eukaryotic RNAs including mRNAs. m 6 A modification has been implicated in mRNA stability and turnover, localization, or translation efficiency. A heterodimeric enzyme complex composed of METTL3 and METTL14 generates m 6 A on mRNAs. METTL3/14 is found in the nucleus where it is localized to nuclear speckles and the splicing regulator WTAP is required for this distinct nuclear localization pattern. Although recent crystal structures revealed how the catalytic MT-A70 domains of METTL3 and METTL14 interact with each other, a more global architecture including WTAP and RNA interactions has not been reported so far. Here, we used recombinant proteins and mapped binding surfaces within the METTL3/14-WTAP complex. Furthermore, we identify nuclear localization signals and identify phosphorylation sites on the endogenous proteins. Using an in vitro methylation assay, we confirm that monomeric METTL3 is soluble and inactive while the catalytic center of METTL14 is degenerated and thus also inactive. In addition, we show that the C-terminal RGG repeats of METTL14 are required for METTL3/14 activity by contributing to RNA substrate binding. Our biochemical work identifies characteristic features of METTL3/14-WTAP and reveals novel insight into the overall architecture of this important enzyme complex.
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A functional nuclear localization signal (NLS) on METTL3 has been reported, and NLS mutant of METTL3 retains in cytoplasm.
“A functional nuclear localization signal (NLS) on METTL3 has been reported, and NLS mutant of METTL3 retains in cytoplasm (Schöller et al., 2018).”
From METTL3 uncouples chromatin accessibility from transcription during retinal development (2025) · cites this paper as Schöller et al., 2018
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METTL3 uncouples chromatin accessibility from transcription during retinal development
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- Authors
- Schöller, E., Weichmann, F., Treiber, T., Ringle, S., Treiber, N., Flatley, A., Feederle, R., Bruckmann, A., & Meister, G.
- Affiliations
- University of Regensburg (DE)
- Helmholtz Munich (DE)
- Corresponding author
- Meister, Gunter
- Venue
- RNA
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 2018-01-18
- Volume
- 24
- Issue
- 4
- Pages
- 499-512
- ISSN
- 1355-8382 (ISSN-L)
- 1469-9001
- PMC
- PMC5855951
- OpenAlex
- W2791366345
- Cited by
- 439
- References
- 5
- Citations by year
- 2026: 38 · 2025: 57 · 2024: 55 · 2023: 68 · 2022: 79 · 2021: 57 · 2020: 50 · 2019: 23 · 2018: 12
- FWCI
- 12.63
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- cc-by-nc
- Locations
- RNA (publishedVersion, cc-by-nc)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- University of Regensburg Publication Server (University of Regensburg) (submittedVersion)
- HighWire Press Open Archive (submittedVersion)
- Site cant be reached (submittedVersion, cc-by)
- PuSH - Publication Server of Helmholtz Zentrum München (submittedVersion, cc-by)
- Funders
- Deutsche Forschungsgemeinschaft
- Bavarian Systems-Biology Network
- European Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cancer-related gene regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- N6-methyladenosine, m6A RNA methylation, METTL3, METTL14, WTAP, protein-protein interaction, nuclear localization signal, protein phosphorylation, nuclear speckles, mRNA stability
- MeSH terms
- RNA Splicing Factors, Adenosine, Amino Acid Sequence, HeLa Cells, Humans, Methyltransferases, Nuclear Proteins, Phosphorylation, Recombinant Proteins, RNA Processing, Post-Transcriptional, RNA, Messenger, RNA-Binding Proteins, Cell Cycle Proteins, Catalytic Domain, Protein Interaction Mapping, Cell Line, Tumor, Multiprotein Complexes, HEK293 Cells
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