Structural analysis of the ribosome nascentpeptide-chain complex of mammalian mitochondria

• Project/Area Number: 18K06054
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43010:Molecular biology-related
• Research Institution: The University of Tokyo
• Principal Investigator: Tomita-Takeuchi Nono 東京大学, 大学院新領域創成科学研究科, 准教授 (80323450)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 翻訳 / 哺乳類ミトコンドリア / リボソーム / 新生ペプチド鎖 / ミトコンドリア / タンパク質合成 / ペプチドトンネル / cryoEM

Outline of Final Research Achievements

This study aims to clarify the structure of the peptide tunnel of the mammalian mitochondrial ribosome and to gain insight into the interaction between the nascent peptide-chain and the peptide tunnel. We have reconstituted the mammalian mitochondrial translation system and successfully formed a ribosome-nascentchain complex(RNC) utilizing translation arrest via a polyproline sequence (Lee, 2021). By examining the length of the nascent peptide-chain, we demonstrated that the N-terminus of the nascent peptide-chain is exposed outside the peptide tunnel when the length of the nascent peptide-chain is more than approximately 80aa.

Academic Significance and Societal Importance of the Research Achievements

近年、ミトコンドリアのリボソームや翻訳因子に誤作用して副作用をもたらす薬剤が多く報告されている。また、リボソームのペプチドトンネルは、新生ペプチド鎖のプロセシングや折りたたみ、細胞内輸送などの制御をになう場であり、ペプチドトンネルは多くの抗生物質の結合部位となっている。本研究により、新生ペプチド鎖の配列や長さを制御しながらミトコンドリアのRNCを調製することが可能になった。今後様々なRNCの構造を決定することにより、薬剤デザイン等に有用な知見が得られると期待される。

Research Products

• [Int'l Joint Research] IMPB, Charite(ドイツ)
• [Int'l Joint Research] IMPB, Charite(ドイツ)
• [Int'l Joint Research] IMPB, Charite(ドイツ)
• [Journal Article] Reconstitution of yeast translation elongation and termination in vitro utilizing CrPV IRES-containing mRNA. (2020)
  • Author(s): Taisho Abe, Riku Nagai, Hiroaki Imataka, and Nono Takeuchi-Tomita
  • Journal Title: J. Biochemistry Volume: - Issue: 5 Pages: 441-450
  • DOI: 10.1093/jb/mvaa021
  • NAID: 40022226784
  • Peer Reviewed / Open Access
• [Journal Article] In vitro yeast reconstituted translation system reveals function of eIF5A for synthesis of long polypeptide (2020)
  • Author(s): Abe Taisho、Nagai Riku、Shimazaki Shunta、Kondo Shunta、Nishimura Satoshi、Sakaguchi Yuriko、Suzuki Tsutomu、Imataka Hiroaki、Tomita Kozo、Takeuchi-Tomita Nono
  • Journal Title: The Journal of Biochemistry Volume: 167 Issue: 5 Pages: 451-462
  • DOI: 10.1093/jb/mvaa022
  • NAID: 40022226787
  • Peer Reviewed / Open Access
• [Journal Article] Reconstitution of mammalian mitochondrial translation system capable of correct initiation and long polypeptide synthesis from leaderless mRNA (2020)
  • Author(s): Lee Muhoon、Matsunaga Noriko、Akabane Shiori、Yasuda Ippei、Ueda Takuya、Takeuchi-Tomita Nono
  • Journal Title: Nucleic Acids Research Volume: 49 Issue: 1 Pages: 371-382
  • DOI: 10.1093/nar/gkaa1165
  • Peer Reviewed / Open Access
• [Presentation] 哺乳類ミトコンドリアタンパク質合成系の試験管内再構成と分子機構 (2021)
  • Author(s): 富田（竹内）野乃
  • Organizer: 日本ミトコンドリア学会年会
  • Int'l Joint Research / Invited
• [Presentation] 哺乳類ミトコンドリアの新生ペプチド鎖リボソーム複合体の機能構造動態 (2019)
  • Author(s): 安田 一平、上田 卓也、富田 野乃
  • Organizer: 第21回日本RNA学会年会
  • Int'l Joint Research
• [Presentation] IRD-like mechanism of polyproline-mediated ribosome stalling and function of eIF5A (2018)
  • Author(s): Nono Takeuchi-Tomita
  • Organizer: 第41回日本分子生物学会年会
  • Int'l Joint Research / Invited
• [Presentation] IRD-like mechanism of polyproline-mediated ribosome stalling and function of eIF5A (2018)
  • Author(s): Nono Takeuchi-Tomita
  • Organizer: Translational control
  • Int'l Joint Research