title: Identification and characterization of a novel evolutionarily conserved lysine-specific methyltransferase targeting eukaryotic translation elongation factor 2 (eEF2).
abstract: The components of the cellular protein translation machinery, such as ribosomal proteins and translation factors, are subject to numerous post-translational modifications. In particular, this group of proteins is frequently methylated. However, for the majority of these methylations, the responsible methyltransferases (MTases) remain unknown. The human FAM86A (family with sequence similarity 86) protein belongs to a recently identified family of protein MTases, and we here show that FAM86A catalyzes the trimethylation of eukaryotic elongation factor 2 (eEF2) on Lys-525. Moreover, we demonstrate that the Saccharomyces cerevisiae MTase Yjr129c, which displays sequence homology to FAM86A, is a functional FAM86A orthologue, modifying the corresponding residue (Lys-509) in yeast eEF2, both in vitro and in vivo. Finally, Yjr129c-deficient yeast cells displayed phenotypes related to eEF2 function (i.e. increased frameshifting during protein translation and hypersensitivity toward the eEF2-specific drug sordarin). In summary, the present study establishes the function of the previously uncharacterized MTases FAM86A and Yjr129c, demonstrating that these enzymes introduce a functionally important lysine methylation in eEF2. Based on the previous naming of similar enzymes, we have redubbed FAM86A and Yjr129c as eEF2-KMT and Efm3, respectively.
authors.affiliation: Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway.
authors.forename: Erna
authors.initials: E
authors.lastname: Davydova
authors.affiliation: Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway.
authors.forename: Angela Y Y
authors.initials: AYY
authors.lastname: Ho
authors.affiliation: Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway.
authors.forename: Jedrzej
authors.initials: J
authors.lastname: Malecki
authors.affiliation: Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway.
authors.forename: Anders
authors.initials: A
authors.lastname: Moen
authors.affiliation: Department of Microbiology, Oslo University Hospital and University of Oslo, 0027 Oslo, Norway, and.
authors.forename: Jorrit M
authors.initials: JM
authors.lastname: Enserink
authors.affiliation: Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway.
authors.forename: Magnus E
authors.initials: ME
authors.lastname: Jakobsson
authors.affiliation: Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford OX1 3TA, United Kingdom.
authors.forename: Christoph
authors.initials: C
authors.lastname: Loenarz
authors.affiliation: Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway,. Electronic address: pal.falnes@ibv.uio.no.
authors.forename: Pål Ø
authors.initials: PØ
authors.lastname: Falnes
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D057809:HEK293 Cells; D006801:Humans; D008745:Methylation; D008780:Methyltransferases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D020652:Peptide Elongation Factor 2 / Q000378:metabolism*; D011496:Protein Methyltransferases / Q000502:physiology*; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D020652:Peptide Elongation Factor 2; D029701:Saccharomyces cerevisiae Proteins; C000594045:EEF2KMT protein, human; D008780:Methyltransferases; D011496:Protein Methyltransferases; C523514:YJR129C protein, S cerevisiae
keywords: Enzyme; Enzyme Catalysis; Post-translational Modification (PTM); Protein Methylation; Translation Elongation Factor