Reconstitution of mammalian mitochondrial translation system and its application

• Project/Area Number: 25440004
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Molecular biology
• Research Institution: The University of Tokyo
• Principal Investigator: Tomita-Takeuchi Nono 東京大学, 新領域創成科学研究科, 准教授 (80323450)
• Research Collaborator: Akabane Shiori 東京大学, 大学院新領域創成科学研究科, 博士課程大学院生
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 翻訳 / ミトコンドリア / 生体外タンパク質合成系 / タンパク質合成

Outline of Final Research Achievements

Overall objective of this study is to elucidate the molecular mechanism of translation system in mammalian mitochondria. As a result, we aim to lay the basis for the medical application, such as the design of antibiotics. Utilizing the reconstituted mitochondrial translation system, we have dissected the function of mitochondrial ICT1, a mitoribosomal protein, and yet a member of peptide release factor family. Its substrate specificity as well as the functional domain has been analyzed. We demonstrated the possible engagement of ICT1 in the translation termination at non-standard stop codons AGA and AGG of two mitochondrial ORFs MTCOI and MTND6, respectively. Structures of the mitoribosomal complexes have been determined by cryoEM reconstruction. The structures of PRE- and POST-translocation complex have been determined at high resolution of approximately 4 angstrom.

Research Products

• [Int'l Joint Research] IMPB, Charite, Berlin(Germany)
• [Int'l Joint Research]
• [Journal Article] Response to the Formal Letter of Z Chrzanowska-Lightowlers and RN Lightowlers regarding our article "Ribosome rescue and translation termination at non-standard stop codons by ICT1 in mammalian mitochondria" (2015)
  • Author(s): Nierhaus, KH. & Takeuchi N.
  • Journal Title: PLoS Genet. Volume: 11(6) Issue: 6 Pages: e1005218-e1005218
  • DOI: 10.1371/journal.pgen.1005218
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Oxidation of translation factor EF-G transiently retards the translational elongation cycle in Escherichia coli (2015)
  • Author(s): Nagano, T., Yutthanasirikul, R., Hihara, Y., Hisabori, T., Kanamori, T., Takeuchi, N., Ueda, T. and Nishiyama, Y.
  • Journal Title: J. Biochem. Volume: - Issue: 2 Pages: 165-72
  • DOI: 10.1093/jb/mvv026
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Ribosome rescue and translation termination at non-standard stop codons by ICT1 in mammalian mitochondria (2014)
  • Author(s): Akabane, S., Ueda, T., Knud H. Nierhaus, KH. & Takeuchi N.
  • Journal Title: PLOS Genetics Volume: 18 Issue: 9 Pages: e1004616-e1004616
  • DOI: 10.1371/journal.pgen.1004616
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Human G-proteins, ObgH1 and Mtg1, associate with the large mitochondrial ribosome subunit and are involved in translation and assembly of respiratory complexes (2013)
  • Author(s): Kotani, T.
  • Journal Title: Nucleic acids research Volume: 2013 Issue: 6 Pages: 3713-3722
  • DOI: 10.1093/nar/gkt079
  • Peer Reviewed
• [Journal Article] Nuclear Respiratory Factor 2beta (NRF-2beta) Recruits NRF-2alpha to the Nucleus by Binding to Importin-alpha:beta via an Unusual Monopartite-Type Nuclear Localization Signal (2013)
  • Author(s): Hayashi, R.
  • Journal Title: Journal of molecular biology Volume: 425 Issue: 18 Pages: 3536-3548
  • DOI: 10.1016/j.jmb.2013.07.007
  • Peer Reviewed
• [Journal Article] 翻訳伸長制御のメカニズムとその意義 (2013)
  • Author(s): 富田(竹内)野乃、上田卓也
  • Journal Title: 実験医学増刊号『生命分子を統合するRNA』 Volume: 31 Pages: 1108-1116
• [Presentation] Unique translation termination in mammalian mitochondria (2015)
  • Author(s): Nono Tomita-Takeuchi
  • Organizer: Molecular Mitochondria
  • Place of Presentation: Helsinki, Finland
  • Year and Date: 2015-10-05
  • Int'l Joint Research / Invited
• [Presentation] 真核細胞におけるnon-canonicalな翻訳制御因子Stm1の機能解析 (2014)
  • Author(s): 林光、上田卓也、伊藤耕一、梶昭、竹内野乃
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県、横浜市）
  • Year and Date: 2014-11-25
• [Presentation] Characterization of eukarytic translation initiation factor 5A from Saccharomyces cerevisiae. (2013)
  • Author(s): Satoshi Nishimura, Hikari Hayashi, Tetsuya Kotani, Takuya Ueda, Koichi Ito, Nono Tomita-Takeuchi
  • Organizer: 2013 RIboclub annual meeting
  • Place of Presentation: ケベック（カナダ）
• [Presentation] The Regulation of elongation Phase (2013)
  • Author(s): Hikari Hayashi 1, Satoshi Nishimura 1, Tetsuya Kotani 1, Syun Utsunomiya 1, Masanari Ishibashi 1, Takuya Ueda, Koichi Ito, Akira Kaji, Nono Tomita-Takeuchi
  • Organizer: 2013 RIboclub annual meeting
  • Place of Presentation: ケベック（カナダ）
• [Presentation] 哺乳類ミトコンドリアにおける異常停止したリボゾーム複合体の再生機構 (2013)
  • Author(s): 赤羽しおり、小谷哲也、入江遼平、上田卓也、竹内野乃
  • Organizer: 第15回日本RNA学会年会
  • Place of Presentation: 愛媛県県民文化会館（日本）
• [Presentation] Protein Synthesis System of Mammalian Mitochondria
  • Author(s): Nono Takeuchi
  • Organizer: ASMRM2013
  • Place of Presentation: ソウル（韓国）
  • Invited