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PMID: 29535189 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Zc3h13/Flacc is required for adenosine methylation by bridging the mRNA-binding factor Rbm15/Spenito to the m6A machinery component Wtap/Fl(2)d.

Genes & development ·Vol. 32 ·No. 5-6 ·2018-00-01 ·Pages 415-429

Knuckles P, Lence T, Haussmann IU, Jacob D, Kreim N, Carl SH, Masiello I, Hares T, Villaseñor R, Hess D, Andrade-Navarro MA, Biggiogera M, Helm M, Soller M, Bühler M, Roignant JY

Abstract

N6-methyladenosine (m6A) is the most abundant mRNA modification in eukaryotes, playing crucial roles in multiple biological processes. m6A is catalyzed by the activity of methyltransferase-like 3 (Mettl3), which depends on additional proteins whose precise functions remain poorly understood. Here we identified Zc3h13 (zinc finger CCCH domain-containing protein 13)/Flacc [Fl(2)d-associated complex component] as a novel interactor of m6A methyltransferase complex components in Drosophila and mice. Like other components of this complex, Flacc controls m6A levels and is involved in sex determination in Drosophila We demonstrate that Flacc promotes m6A deposition by bridging Fl(2)d to the mRNA-binding factor Nito. Altogether, our work advances the molecular understanding of conservation and regulation of the m6A machinery.

Keywords
Flacc RNA modifications Zc3h13 m6A methyltransferase complex sex determination
MeSH Terms
Adenosine/metabolism Animals Carrier Proteins/metabolism Cell Cycle Proteins Cell Line DNA-Binding Proteins/metabolism Drosophila Proteins/metabolism Drosophila melanogaster/enzymology,physiology Gene Expression Regulation, Developmental Methylation Methyltransferases/metabolism Mice Mouse Embryonic Stem Cells Nuclear Proteins/metabolism Protein Transport RNA Precursors/genetics RNA Splicing RNA Splicing Factors RNA-Binding Proteins/metabolism Sex Determination Processes/genetics
Chemicals
Carrier Proteins Cell Cycle Proteins DNA-Binding Proteins Drosophila Proteins NITO protein, Drosophila Nuclear Proteins RNA Precursors RNA Splicing Factors RNA-Binding Proteins Rbm15 protein, mouse Wtap protein, mouse Zc3h13 protein, mouse Methyltransferases Mettl3 protein, mouse Adenosine
Authors & Affiliations
16 authors, click to expand affiliations / ORCID
Knuckles Philip
Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland. | University of Basel, Basel 4002, Switzerland.
Lence Tina
Institute of Molecular Biology, 55128 Mainz, Germany.
Haussmann Irmgard U
School of Life Science, Faculty of Health and Life Sciences, Coventry University, Coventry CV1 5FB, United Kingdom. | School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
Jacob Dominik
Institute of Pharmacy and Biochemistry, Johannes Gutenberg University of Mainz, 55128 Mainz, Germany.
Kreim Nastasja
Bioinformatics Core Facility, Institute of Molecular Biology, 55128 Mainz, Germany.
Carl Sarah H
Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland. | Swiss Institute of Bioinformatics, Basel 4058, Switzerland.
Masiello Irene
Institute of Molecular Biology, 55128 Mainz, Germany. | Laboratory of Cell Biology and Neurobiology, Department of Animal Biology, University of Pavia, Pavia 27100, Italy.
Hares Tina
Institute of Molecular Biology, 55128 Mainz, Germany.
Villaseñor Rodrigo
Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland. | University of Basel, Basel 4002, Switzerland.
Hess Daniel
Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland.
Andrade-Navarro Miguel A
Institute of Molecular Biology, 55128 Mainz, Germany. | Faculty of Biology, Johannes Gutenberg University of Mainz, 55128 Mainz, Germany.
Biggiogera Marco
Laboratory of Cell Biology and Neurobiology, Department of Animal Biology, University of Pavia, Pavia 27100, Italy.
Helm Mark
Institute of Pharmacy and Biochemistry, Johannes Gutenberg University of Mainz, 55128 Mainz, Germany.
Soller Matthias
School of Life Science, Faculty of Health and Life Sciences, Coventry University, Coventry CV1 5FB, United Kingdom.
Bühler Marc
Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland. | University of Basel, Basel 4002, Switzerland.
Roignant Jean-Yves
Institute of Molecular Biology, 55128 Mainz, Germany.
References (48)
48 references, click to expand
  1. Single-Step Generation of Conditional Knockout Mouse Embryonic Stem Cells.
    Cell Rep. 2015 Jul 28;12(4):709-16 PMID: 26190102
  2. RNA fate determination through cotranscriptional adenosine methylation and microprocessor binding.
    Nat Struct Mol Biol. 2017 Jul;24(7):561-569 PMID: 28581511
  3. spenito is required for sex determination in Drosophila melanogaster.
    Proc Natl Acad Sci U S A. 2015 Sep 15;112(37):11606-11 PMID: 26324914
  4. Absolute and relative quantification of RNA modifications via biosynthetic isotopomers.
    Nucleic Acids Res. 2014 Oct;42(18):e142 PMID: 25129236
  5. Comprehensive analysis of mRNA methylation reveals enrichment in 3' UTRs and near stop codons.
    Cell. 2012 Jun 22;149(7):1635-46 PMID: 22608085
  6. Purification and cDNA cloning of the AdoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase.
    RNA. 1997 Nov;3(11):1233-47 PMID: 9409616
  7. Reader interactome of epigenetic histone marks in birds.
    Proteomics. 2016 Feb;16(3):427-36 PMID: 26703087
  8. Temporal Control of Mammalian Cortical Neurogenesis by m6A Methylation.
    Cell. 2017 Nov 2;171(4):877-889.e17 PMID: 28965759
  9. NanoLuc Complementation Reporter Optimized for Accurate Measurement of Protein Interactions in Cells.
    ACS Chem Biol. 2016 Feb 19;11(2):400-8 PMID: 26569370
  10. Basal forebrain in the context of schizophrenia.
    Brain Res Brain Res Rev. 2000 Mar;31(2-3):205-35 PMID: 10719150
  11. Panorama of ancient metazoan macromolecular complexes.
    Nature. 2015 Sep 17;525(7569):339-44 PMID: 26344197
  12. Structural basis of N(6)-adenosine methylation by the METTL3-METTL14 complex.
    Nature. 2016 May 25;534(7608):575-8 PMID: 27281194
  13. The m6A pathway facilitates sex determination in Drosophila.
    Nat Commun. 2017 Jul 04;8:15737 PMID: 28675155
  14. m6A RNA Methylation Regulates the Self-Renewal and Tumorigenesis of Glioblastoma Stem Cells.
    Cell Rep. 2017 Mar 14;18(11):2622-2634 PMID: 28297667
  15. Accurate proteome-wide label-free quantification by delayed normalization and maximal peptide ratio extraction, termed MaxLFQ.
    Mol Cell Proteomics. 2014 Sep;13(9):2513-26 PMID: 24942700
  16. Genetic alterations of m6A regulators predict poorer survival in acute myeloid leukemia.
    J Hematol Oncol. 2017 Feb 2;10 (1):39 PMID: 28153030
  17. Identification of Wilms' tumor 1-associating protein complex and its role in alternative splicing and the cell cycle.
    J Biol Chem. 2013 Nov 15;288(46):33292-302 PMID: 24100041
  18. Combined analysis of exon splicing and genome wide polymorphism data predict schizophrenia risk loci.
    J Psychiatr Res. 2014 May;52:44-9 PMID: 24507884
  19. Structural Basis for Cooperative Function of Mettl3 and Mettl14 Methyltransferases.
    Mol Cell. 2016 Jul 21;63(2):306-317 PMID: 27373337
  20. A fly view on the roles and mechanisms of the m6A mRNA modification and its players.
    RNA Biol. 2017 Sep 2;14 (9):1232-1240 PMID: 28353398
  21. A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation.
    Nat Chem Biol. 2014 Feb;10(2):93-5 PMID: 24316715
  22. The gene virilizer is required for female-specific splicing controlled by Sxl, the master gene for sexual development in Drosophila.
    Development. 1995 Dec;121(12):4017-26 PMID: 8575302
  23. RNA methylation by the MIS complex regulates a cell fate decision in yeast.
    PLoS Genet. 2012;8(6):e1002732 PMID: 22685417
  24. Identification of factors required for m6 A mRNA methylation in Arabidopsis reveals a role for the conserved E3 ubiquitin ligase HAKAI.
    New Phytol. 2017 Jul;215(1):157-172 PMID: 28503769
  25. Quantitative proteomics combined with BAC TransgeneOmics reveals in vivo protein interactions.
    J Cell Biol. 2010 May 17;189(4):739-54 PMID: 20479470
  26. N6-methyladenosine modification destabilizes developmental regulators in embryonic stem cells.
    Nat Cell Biol. 2014 Feb;16(2):191-8 PMID: 24394384
  27. AAV-mediated direct in vivo CRISPR screen identifies functional suppressors in glioblastoma.
    Nat Neurosci. 2017 Oct;20(10 ):1329-1341 PMID: 28805815
  28. m6A modulates neuronal functions and sex determination in Drosophila.
    Nature. 2016 Dec 8;540(7632):242-247 PMID: 27919077
  29. deepTools2: a next generation web server for deep-sequencing data analysis.
    Nucleic Acids Res. 2016 Jul 8;44(W1):W160-5 PMID: 27079975
  30. Structural insights into the molecular mechanism of the m(6)A writer complex.
    Elife. 2016 Sep 14;5: PMID: 27627798
  31. The N6-methyladenosine (m6A)-forming enzyme METTL3 controls myeloid differentiation of normal hematopoietic and leukemia cells.
    Nat Med. 2017 Nov;23 (11):1369-1376 PMID: 28920958
  32. Global quantification of mammalian gene expression control.
    Nature. 2011 May 19;473(7347):337-42 PMID: 21593866
  33. Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq.
    Nature. 2012 Apr 29;485(7397):201-6 PMID: 22575960
  34. m6A Demethylase ALKBH5 Maintains Tumorigenicity of Glioblastoma Stem-like Cells by Sustaining FOXM1 Expression and Cell Proliferation Program.
    Cancer Cell. 2017 Apr 10;31(4):591-606.e6 PMID: 28344040
  35. Andromeda: a peptide search engine integrated into the MaxQuant environment.
    J Proteome Res. 2011 Apr 1;10(4):1794-805 PMID: 21254760
  36. m(6)A RNA methylation promotes XIST-mediated transcriptional repression.
    Nature. 2016 Sep 15;537(7620):369-373 PMID: 27602518
  37. Perturbation of m6A writers reveals two distinct classes of mRNA methylation at internal and 5' sites.
    Cell Rep. 2014 Jul 10;8(1):284-96 PMID: 24981863
  38. METTL14 Inhibits Hematopoietic Stem/Progenitor Differentiation and Promotes Leukemogenesis via mRNA m6A Modification.
    Cell Stem Cell. 2018 Feb 1;22(2):191-205.e9 PMID: 29290617
  39. The Perseus computational platform for comprehensive analysis of (prote)omics data.
    Nat Methods. 2016 Sep;13(9):731-40 PMID: 27348712
  40. Nucleotide modifications in messenger RNA and their role in development and disease.
    Biochem Soc Trans. 2016 Oct 15;44(5):1385-1393 PMID: 27911721
  41. m6A in mRNA: An Ancient Mechanism for Fine-Tuning Gene Expression.
    Trends Genet. 2017 Jun;33(6):380-390 PMID: 28499622
  42. m6A potentiates Sxl alternative pre-mRNA splicing for robust Drosophila sex determination.
    Nature. 2016 Dec 8;540(7632):301-304 PMID: 27919081
  43. Significance analysis of microarrays applied to the ionizing radiation response.
    Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5116-21 PMID: 11309499
  44. MTA is an Arabidopsis messenger RNA adenosine methylase and interacts with a homolog of a sex-specific splicing factor.
    Plant Cell. 2008 May;20(5):1278-88 PMID: 18505803
  45. Mammalian WTAP is a regulatory subunit of the RNA N6-methyladenosine methyltransferase.
    Cell Res. 2014 Feb;24(2):177-89 PMID: 24407421
  46. FTO Plays an Oncogenic Role in Acute Myeloid Leukemia as a N6-Methyladenosine RNA Demethylase.
    Cancer Cell. 2017 Jan 9;31(1):127-141 PMID: 28017614
  47. Promoter-bound METTL3 maintains myeloid leukaemia by m6A-dependent translation control.
    Nature. 2017 Dec 7;552(7683):126-131 PMID: 29186125
  48. Emerging links between m6A and misregulated mRNA methylation in cancer.
    Genome Med. 2017 Jan 12;9(1):2 PMID: 28081722
Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
1549-5477
DOI
10.1101/gad.309146.117
Published
2018-00-01
Epub
2018-00-13
Pages
415-429
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC5900714
Subset
IM
External Links
PubMed source DOI: 10.1101/gad.309146.117 PMC Full Text View journal details
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