title: Deficient methylation and formylation of mt-tRNA(Met) wobble cytosine in a patient carrying mutations in NSUN3.
abstract: Epitranscriptome modifications are required for structure and function of RNA and defects in these pathways have been associated with human disease. Here we identify the RNA target for the previously uncharacterized 5-methylcytosine (m(5)C) methyltransferase NSun3 and link m(5)C RNA modifications with energy metabolism. Using whole-exome sequencing, we identified loss-of-function mutations in NSUN3 in a patient presenting with combined mitochondrial respiratory chain complex deficiency. Patient-derived fibroblasts exhibit severe defects in mitochondrial translation that can be rescued by exogenous expression of NSun3. We show that NSun3 is required for deposition of m(5)C at the anticodon loop in the mitochondrially encoded transfer RNA methionine (mt-tRNA(Met)). Further, we demonstrate that m(5)C deficiency in mt-tRNA(Met) results in the lack of 5-formylcytosine (f(5)C) at the same tRNA position. Our findings demonstrate that NSUN3 is necessary for efficient mitochondrial translation and reveal that f(5)C in human mitochondrial RNA is generated by oxidative processing of m(5)C.
authors.affiliation: MRC Mitochondrial Biology Unit, Hills Road, Cambridge CB2 0XY, UK.
authors.forename: Lindsey
authors.initials: L
authors.lastname: Van Haute
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute, Genetics, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
authors.forename: Sabine
authors.initials: S
authors.lastname: Dietmann
authors.affiliation: Helmholtz Zentrum München, Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH), Institute of Human Genetics, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.|Technical University Munich, Institute of Human Genetics, Trogerstrasse 32, 81675 München, Germany.
authors.forename: Laura
authors.initials: L
authors.lastname: Kremer
authors.affiliation: Department of Biology &Biochemistry, University of Bath, Claverton Down, Bath, BA2 7AY, UK.
authors.forename: Shobbir
authors.initials: S
authors.lastname: Hussain
authors.affiliation: MRC Mitochondrial Biology Unit, Hills Road, Cambridge CB2 0XY, UK.
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Pearce
authors.affiliation: MRC Mitochondrial Biology Unit, Hills Road, Cambridge CB2 0XY, UK.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Powell
authors.affiliation: MRC Mitochondrial Biology Unit, Hills Road, Cambridge CB2 0XY, UK.
authors.forename: Joanna
authors.initials: J
authors.lastname: Rorbach
authors.affiliation: Department of Physiology Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3DY, UK.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Lantaff
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute, Genetics, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
authors.forename: Sandra
authors.initials: S
authors.lastname: Blanco
authors.affiliation: Max-Planck-Institute for Molecular Genetics, Otto-Warburg Laboratory, 14195 Berlin, Germany.|University of Würzburg, CU Systems Medicine, 97080 Würzburg, Germany.|Max-Delbrück-Center for Molecular Medicine, Berlin Institute for Medical Systems Biology/Berlin Institute of Health, 13125 Berlin, Germany.
authors.forename: Sascha
authors.initials: S
authors.lastname: Sauer
authors.affiliation: Department of General Pediatrics, Division of Inherited Metabolic Diseases, University Children's Hospital, 69120 Heidelberg, Germany.
authors.forename: Urania
authors.initials: U
authors.lastname: Kotzaeridou
authors.affiliation: Department of General Pediatrics, Division of Inherited Metabolic Diseases, University Children's Hospital, 69120 Heidelberg, Germany.
authors.forename: Georg F
authors.initials: GF
authors.lastname: Hoffmann
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton CB10 1SA, UK.
authors.forename: Yasin
authors.initials: Y
authors.lastname: Memari
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton CB10 1SA, UK.
authors.forename: Anja
authors.initials: A
authors.lastname: Kolb-Kokocinski
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton CB10 1SA, UK.
authors.forename: Richard
authors.identifier: 0000-0002-9130-1006
authors.initials: R
authors.lastname: Durbin
authors.affiliation: Department of Paediatrics, Paracelsus Medical University, SALK Salzburg, Salzburg 5020, Austria.
authors.forename: Johannes A
authors.initials: JA
authors.lastname: Mayr
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute, Genetics, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
authors.forename: Michaela
authors.initials: M
authors.lastname: Frye
authors.affiliation: Helmholtz Zentrum München, Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH), Institute of Human Genetics, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.|Technical University Munich, Institute of Human Genetics, Trogerstrasse 32, 81675 München, Germany.
authors.forename: Holger
authors.identifier: 0000-0003-2379-6286
authors.initials: H
authors.lastname: Prokisch
authors.affiliation: MRC Mitochondrial Biology Unit, Hills Road, Cambridge CB2 0XY, UK.
authors.forename: Michal
authors.initials: M
authors.lastname: Minczuk
mesh_terms: D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008745:Methylation; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D028361:Mitochondrial Diseases / Q000235:genetics*; D009154:Mutation; D012343:RNA, Transfer / Q000378:metabolism*
chemical_list: D012343:RNA, Transfer; D008780:Methyltransferases; C000620888:NSUN3 protein, human