Mammalian NSUN2 introduces 5-methylcytidines into mitochondrial tRNAs
Copyright policy )Mammalian NSUN2 introduces 5-methylcytidines into mitochondrial tRNAs
ABSTRACT Post-transcriptional modifications in mitochondrial tRNAs (mt-tRNAs) play critical roles in mitochondrial protein synthesis, which produces respiratory chain complexes. In this study, we used mass spectrometric analysis to map 5-methylcytidine (m 5 C) at positions 48–50 in eight mouse and six human mt-tRNAs. We also confirmed the absence of m 5 C in mt-tRNAs isolated from Nsun2 knockout (KO) mice, as well as from NSUN2 KO human culture cells. In addition, we successfully reconstituted m 5 C at positions 48–50 of mt-tRNA in vitro with NSUN2 protein in the presence of S -adenosylmethionine (SAM). Although NSUN2 is predominantly localized to the nucleus and introduces m 5 C into cytoplasmic tRNAs and mRNAs, structured illumination microscopy (SIM) clearly revealed NSUN2 foci inside mitochondria. These observations provide novel insights into the role of NSUN2 in the physiology and pathology of mitochondrial functions.
- Hokkaido Bunkyo University Japan
- University of Tokyo Japan
- Kumamoto University Japan
- Hokkeido University Japan
- Kumamoto University
Gene Editing, Mice, Knockout, Primary Cell Culture, Methyltransferases, Fibroblasts, Methylation, Oxidative Phosphorylation, Mitochondria, Gene Knockout Techniques, Mice, Protein Transport, HEK293 Cells, RNA and RNA-protein complexes, 5-Methylcytosine, Animals, Humans, Nucleic Acid Conformation, RNA, Messenger, CRISPR-Cas Systems, RNA Processing, Post-Transcriptional, HeLa Cells
Gene Editing, Mice, Knockout, Primary Cell Culture, Methyltransferases, Fibroblasts, Methylation, Oxidative Phosphorylation, Mitochondria, Gene Knockout Techniques, Mice, Protein Transport, HEK293 Cells, RNA and RNA-protein complexes, 5-Methylcytosine, Animals, Humans, Nucleic Acid Conformation, RNA, Messenger, CRISPR-Cas Systems, RNA Processing, Post-Transcriptional, HeLa Cells
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