Schoeller, Eva and Weichmann, Franziska and Treiber, Thomas and Ringle, Sam and Treiber, Nora and Flatley, Andrew and Feederle, Regina and Bruckmann, Astrid and Meister, Gunter (2018) Interactions, localization, and phosphorylation of the m(6)A generating METTL3-METTL14-WTAP complex. RNA, 24 (4). pp. 499-512. ISSN 1355-8382, 1469-9001
Full text not available from this repository. (Request a copy)Abstract
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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | GENE-EXPRESSION REGULATION; MESSENGER-RNA METHYLATION; N-6-ADENOSINE METHYLATION; NUCLEAR-RNA; TRANSLATION; PROTEIN; N6-METHYLADENOSINE; METHYLTRANSFERASE; IDENTIFICATION; DEMETHYLASE; METTL14; METTL3; RNA modification; m(6)A; methyltransferase |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I > Prof. Dr. Gunter Meister |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 20 Feb 2020 13:58 |
| Last Modified: | 20 Feb 2020 13:58 |
| URI: | https://pred.uni-regensburg.de/id/eprint/14771 |
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