Structural Basis for Cooperative Function of Mettl3 and Mettl14 Methyltransferases

Wang, Ping; Doxtader, Katelyn A.; Nam, Yunsun

doi:10.1016/j.molcel.2016.05.041

Cited by 949 publications

(1,137 citation statements)

References 46 publications

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“…More importantly, in vitro studies show that METTL3 and METTL14 interact directly and stabilize each other at the protein levels (11,34), and increasing evidences suggest that METTL3 is the real catalytically active subunit while METTL14 plays a structural role critical for substrate recognition (35). There also exists evidence that WTAP functions as a regulatory subunit in the m 6 A methyltransferase complex and has a direct interaction with METTL3 (12).…”

Section: Resultsmentioning

confidence: 99%

“…Recent studies have stated the methyltransferase domain (MTD, residues 369–590) and the two Cys-Cys-Cys-His (CCCH)-type zinc finger motifs (ZnF, 259–340) of METTL3 along with METTL14 MTD are necessary for RNA m 6 A modification in vitro methylation activity assays (46). Though the crystal structure of the core METTL3-METTL14 complex comprising the MTase domains have already been elucidated (47), the structure of full-length METTL3 still remains elusive.…”

Section: Discussionmentioning

confidence: 99%

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SUMOylation of the m6A-RNA methyltransferase METTL3 modulates its function

Hou

Zhang

et al. 2018

Nucleic Acids Research

242

176

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The methyltransferase like 3 (METTL3) is a key component of the large N6-adenosine-methyltransferase complex in mammalian responsible for N6-methyladenosine (m6A) modification in diverse RNAs including mRNA, tRNA, rRNA, small nuclear RNA, microRNA precursor and long non-coding RNA. However, the characteristics of METTL3 in activation and post-translational modification (PTM) is seldom understood. Here we find that METTL3 is modified by SUMO1 mainly at lysine residues K177, K211, K212 and K215, which can be reduced by an SUMO1-specific protease SENP1. SUMOylation of METTL3 does not alter its stability, localization and interaction with METTL14 and WTAP, but significantly represses its m6A methytransferase activity resulting in the decrease of m6A levels in mRNAs. Consistently with this, the abundance of m6A in mRNAs is increased with re-expression of the mutant METTL3-4KR compared to that of wild-type METTL3 in human non-small cell lung carcinoma (NSCLC) cell line H1299-shMETTL3, in which endogenous METTL3 was knockdown. The alternation of m6A in mRNAs and subsequently change of gene expression profiles, which are mediated by SUMOylation of METTL3, may directly influence the soft-agar colony formation and xenografted tumor growth of H1299 cells. Our results uncover an important mechanism for SUMOylation of METTL3 regulating its m6A RNA methyltransferase activity.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

SUMOylation of the m6A-RNA methyltransferase METTL3 modulates its function

Hou

Zhang

et al. 2018

Nucleic Acids Research

242

176

View full text Add to dashboard Cite

show abstract

“…The most striking parallel with the prozyme paradigm emerged from the work on the human RNA methyltransferase complex METTL3-METTL14. In this complex, METTL3 constitutes the catalytic core and binds AdoMet but requires allosteric activation and stabilization by METTL14 (48,49). Although METTL14 has been reported to exhibit weak in vitro methylation activity (50), the phylogenetic analysis suggests that the METTL14 catalytic core has lost its function (51).…”

Section: Discussionmentioning

confidence: 99%

The Major Protein Arginine Methyltransferase in Trypanosoma brucei Functions as an Enzyme-Prozyme Complex

Kafková

Debler

Fisk

et al. 2017

Journal of Biological Chemistry

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Edited by John M. DenuProzymes are catalytically inactive enzyme paralogs that dramatically stimulate the function of weakly active enzymes through complex formation. The two prozymes described to date reside in the polyamine biosynthesis pathway of the human parasite Trypanosoma brucei, an early branching eukaryote that lacks transcriptional regulation and regulates its proteome through posttranscriptional and posttranslational means. Arginine methylation is a common posttranslational modification in eukaryotes catalyzed by protein arginine methyltransferases (PRMTs) that are typically thought to function as homodimers. We demonstrate that a major T. brucei PRMT, TbPRMT1, functions as a heterotetrameric enzyme-prozyme pair. The inactive PRMT paralog, TbPRMT1 PRO , is essential for catalytic activity of the TbPRMT1 ENZ subunit. Mutational analysis definitively demonstrates that TbPRMT1 ENZ is the cofactor-binding subunit and carries all catalytic activity of the complex. Our results are the first demonstration of an obligate heteromeric PRMT, and they suggest that enzyme-prozyme organization is expanded in trypanosomes as a posttranslational means of enzyme regulation.

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“…To create a catalytic dead mutant Mettl3 , we inserted the mutations D395A and W398A (DPPW catalytic motif of METTL3) based on published data, which show that those two residues are absolutely required for METTL3 activity 35–37 . Mettl3 KO naïve T cells were isolated from Mettl3 KO mice with StemCell Naïve T cell purification kit, then electroporated by nucleofection by exactly following the Amaxa mouse T cell nucleofector kit manual (Lonza).…”

Section: Methodsmentioning

confidence: 99%

m6A mRNA methylation controls T cell homeostasis by targeting the IL-7/STAT5/SOCS pathways

Tong

Zhu

et al. 2017

Nature

746

820

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N6 -methyladenosine (m6A) is the most common and abundant messenger RNA modification, modulated by ‘writers’, ‘erasers’ and ‘readers’ of this mark 1,2. In vitro data have shown that m6A influences all fundamental aspects of mRNA metabolism, mainly mRNA stability, to determine stem cell fates 3,4. However, its in vivo physiological function in mammals and adult mammalian cells is still unknown. Here we show that deletion of m6A ‘writer’ protein METTL3 in mouse T cells disrupts T cell homeostasis and differentiation. In a lymphopenic mouse adoptive transfer model, naive Mettl3 deficient T cells failed to undergo homeostatic expansion and remarkably remained in the naïve state up through 12 weeks, thereby preventing colitis. Consistent with these observations, the mRNAs of SOCS family genes encoding STAT- signaling inhibitory proteins, Socs1, Socs3 and Cish, were marked by m6A, exhibited slower mRNA decay and increased mRNAs and protein expression levels in Mettl3 deficient naïve T cells. This increased SOCS family activity consequently inhibited IL-7 mediated STAT5 activation and T cell homeostatic proliferation and differentiation. We also found that m6A plays important roles for inducible degradation of Socs mRNAs in response to IL-7 signaling in order to reprogram Naïve T cells for proliferation and differentiation. Our study elucidates for the first time the in vivo biological role of m6A modification in T cell mediated pathogenesis and reveals a novel mechanism of T cell homeostasis and signal-dependent induction of mRNA degradation.

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Structural Basis for Cooperative Function of Mettl3 and Mettl14 Methyltransferases

Cited by 949 publications

References 46 publications

SUMOylation of the m6A-RNA methyltransferase METTL3 modulates its function

SUMOylation of the m6A-RNA methyltransferase METTL3 modulates its function

The Major Protein Arginine Methyltransferase in Trypanosoma brucei Functions as an Enzyme-Prozyme Complex

m6A mRNA methylation controls T cell homeostasis by targeting the IL-7/STAT5/SOCS pathways

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