NSUN2 introduces 5-methylcytosines in mammalian mitochondrial tRNAs.
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Authors
Van Haute, Lindsey
McCann, Beverly J
Powell, Christopher A
Bansal, Dhiru
Vasiliauskaitė, Lina
Garone, Caterina
Shin, Sanghee
Gleeson, Joseph G
Rhee, Hyun-Woo
Publication Date
2019-09Journal Title
Nucleic acids research
ISSN
0305-1048
Publisher
Oxford University Press
Volume
47
Issue
16
Pages
8720-8733
Language
eng
Type
Article
This Version
VoR
Physical Medium
Print
Metadata
Show full item recordCitation
Van Haute, L., Lee, S., McCann, B. J., Powell, C. A., Bansal, D., Vasiliauskaitė, L., Garone, C., et al. (2019). NSUN2 introduces 5-methylcytosines in mammalian mitochondrial tRNAs.. Nucleic acids research, 47 (16), 8720-8733. https://doi.org/10.1093/nar/gkz559
Abstract
Expression of human mitochondrial DNA is indispensable for proper function of the oxidative phosphorylation machinery. The mitochondrial genome encodes 22 tRNAs, 2 rRNAs and 11 mRNAs and their post-transcriptional modification constitutes one of the key regulatory steps during mitochondrial gene expression. Cytosine-5 methylation (m5C) has been detected in mitochondrial transcriptome, however its biogenesis has not been investigated in details. Mammalian NOP2/Sun RNA Methyltransferase Family Member 2 (NSUN2) has been characterized as an RNA methyltransferase introducing m5C in nuclear-encoded tRNAs, mRNAs and microRNAs and associated with cell proliferation and differentiation, with pathogenic variants in NSUN2 being linked to neurodevelopmental disorders. Here we employ spatially restricted proximity labelling and immunodetection to demonstrate that NSUN2 is imported into the matrix of mammalian mitochondria. Using three genetic models for NSUN2 inactivation-knockout mice, patient-derived fibroblasts and CRISPR/Cas9 knockout in human cells-we show that NSUN2 is necessary for the generation of m5C at positions 48, 49 and 50 of several mammalian mitochondrial tRNAs. Finally, we show that inactivation of NSUN2 does not have a profound effect on mitochondrial tRNA stability and oxidative phosphorylation in differentiated cells. We discuss the importance of the newly discovered function of NSUN2 in the context of human disease.
Keywords
Mitochondria, Fibroblasts, Animals, Mice, Knockout, Humans, Mice, Microcephaly, Eczema, Facies, Growth Disorders, 5-Methylcytosine, Methyltransferases, RNA, Messenger, RNA, Transfer, RNA Processing, Post-Transcriptional, Nucleic Acid Conformation, Methylation, Protein Transport, Oxidative Phosphorylation, Gene Knockout Techniques, HEK293 Cells, Intellectual Disability, Primary Cell Culture, CRISPR-Cas Systems, Gene Editing, RNA, Mitochondrial
Sponsorship
Medical Research Council, UK [MC_UU_00015/4 to M.M.]; EMBO [ALFT 701-2013 to L.V.H.]; National Research Foundation of Korea [NRF-2019R1A2C3008463 to S.Y.L and H.W.R.]; Cancer Research UK [C13474/A18583, C6946/A14492 to E.A.M.]; Wellcome Trust [104640/Z/14/Z, 092096/Z/10/Z to E.A.M.]. Funding for open access charge: MRC.
Funder references
European Commission Horizon 2020 (H2020) Marie Sk?odowska-Curie actions (721757)
MRC (MC_U105697135)
MRC (MC_UU_00015/4)
Wellcome Trust (092096/Z/10/Z)
WELLCOME TRUST (104640/Z/14/Z)
Cancer Research UK (18583)
Cancer Research UK (A14492)
Identifiers
External DOI: https://doi.org/10.1093/nar/gkz559
This record's URL: https://www.repository.cam.ac.uk/handle/1810/296162