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Mammalian NSUN2 introduces 5-methylcytidines into mitochondrial tRNAs

Saori Shinoda, Sho Kitagawa, View ORCID ProfileShinichi Nakagawa, Fan-Yan Wei, Kazuhito Tomizawa, Kimi Araki, Masatake Araki, View ORCID ProfileTakeo Suzuki, Tsutomu Suzuki
doi: https://doi.org/10.1101/624932
Saori Shinoda
1Department of Chemistry and Biotechnology, Graduate School of Engineering, the University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan
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Sho Kitagawa
1Department of Chemistry and Biotechnology, Graduate School of Engineering, the University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan
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Shinichi Nakagawa
2Faculty of Pharmaceutical Sciences, Hokkaido University, Hokkaido 060-0812, Japan
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  • ORCID record for Shinichi Nakagawa
Fan-Yan Wei
3Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto 860-8556, Japan
4Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Kawaguchi, Japan
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Kazuhito Tomizawa
3Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto 860-8556, Japan
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Kimi Araki
5Institute of Resource Development and Analysis, Kumamoto University, Kumamoto 860-0811, Japan
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Masatake Araki
5Institute of Resource Development and Analysis, Kumamoto University, Kumamoto 860-0811, Japan
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Takeo Suzuki
1Department of Chemistry and Biotechnology, Graduate School of Engineering, the University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan
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  • ORCID record for Takeo Suzuki
  • For correspondence: ts@chembio.t.u-tokyo.ac.jp t_suzuki@chembio.t.u-tokyo.ac.jp
Tsutomu Suzuki
1Department of Chemistry and Biotechnology, Graduate School of Engineering, the University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan
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  • For correspondence: ts@chembio.t.u-tokyo.ac.jp t_suzuki@chembio.t.u-tokyo.ac.jp
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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 (m5C) at positions 48–50 in eight mouse and six human mt-tRNAs. We also confirmed the absence of m5C in mt-tRNAs isolated from Nsun2 knockout (KO) mice, as well as from NSUN2 KO human culture cells. In addition, we successfully reconstituted m5C 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 m5C 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.

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Posted May 02, 2019.
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Mammalian NSUN2 introduces 5-methylcytidines into mitochondrial tRNAs
Saori Shinoda, Sho Kitagawa, Shinichi Nakagawa, Fan-Yan Wei, Kazuhito Tomizawa, Kimi Araki, Masatake Araki, Takeo Suzuki, Tsutomu Suzuki
bioRxiv 624932; doi: https://doi.org/10.1101/624932
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Mammalian NSUN2 introduces 5-methylcytidines into mitochondrial tRNAs
Saori Shinoda, Sho Kitagawa, Shinichi Nakagawa, Fan-Yan Wei, Kazuhito Tomizawa, Kimi Araki, Masatake Araki, Takeo Suzuki, Tsutomu Suzuki
bioRxiv 624932; doi: https://doi.org/10.1101/624932

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