title: N6-methyladenosine modification destabilizes developmental regulators in embryonic stem cells.
abstract: N(6)-methyladenosine (m(6)A) has been identified as the most abundant internal modification of messenger RNA in eukaryotes. m(6)A modification is involved in cell fate determination in yeast and embryo development in plants. Its mammalian function remains unknown but thousands of mammalian mRNAs and long non-coding RNAs (lncRNAs) show m(6)A modification and m(6)A demethylases are required for mammalian energy homeostasis and fertility. We identify two proteins, the putative m(6)A MTase, methyltransferase-like 3 (Mettl3; ref. ), and a related but uncharacterized protein Mettl14, that function synergistically to control m(6)A formation in mammalian cells. Knockdown of Mettl3 and Mettl14 in mouse embryonic stem cells (mESCs) led to similar phenotypes, characterized by lack of m(6)A RNA methylation and lost self-renewal capability. A large number of transcripts, including many encoding developmental regulators, exhibit m(6)A methylation inversely correlated with mRNA stability and gene expression. The human antigen R (HuR) and microRNA pathways were linked to these effects. This gene regulatory mechanism operating in mESCs through m(6)A methylation is required to keep mESCs at their ground state and may be relevant to thousands of mRNAs and lncRNAs in various cell types.
authors.affiliation: Tumor Initiation and Maintenance Program, NCI-designated Cancer Center, Sanford Burnham Medical Research Institute, La Jolla, California 92037, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Computer Science, The Donnelly Centre, University of Toronto, Toronto M5S 3G4, Canada.
authors.forename: Yue
authors.initials: Y
authors.lastname: Li
authors.affiliation: Tumor Initiation and Maintenance Program, NCI-designated Cancer Center, Sanford Burnham Medical Research Institute, La Jolla, California 92037, USA.
authors.forename: Julia I
authors.initials: JI
authors.lastname: Toth
authors.affiliation: Tumor Initiation and Maintenance Program, NCI-designated Cancer Center, Sanford Burnham Medical Research Institute, La Jolla, California 92037, USA.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Petroski
authors.affiliation: 1] Department of Computer Science, The Donnelly Centre, University of Toronto, Toronto M5S 3G4, Canada [2] Department of Molecular Genetics, Donnelly Centre for Cellular and Biomolecular Research, Banting and Best Department of Medical Research, University of Toronto, Toronto M5S 3G4, Canada.
authors.forename: Zhaolei
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Tumor Initiation and Maintenance Program, NCI-designated Cancer Center, Sanford Burnham Medical Research Institute, La Jolla, California 92037, USA.
authors.forename: Jing Crystal
authors.initials: JC
authors.lastname: Zhao
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D006801:Humans; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D017386:Sequence Homology, Amino Acid
chemical_list: D012333:RNA, Messenger; C010223:N-methyladenosine; C587355:METTL14 protein, human; D008780:Methyltransferases; C000596366:Mettl3 protein, mouse; C468275:METTL3 protein, human; D000241:Adenosine