Molecular mechanism of ribosome hijack by the HCV IRES

• Project/Area Number: 16H04756
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Ito Takuhiro 国立研究開発法人理化学研究所, 生命機能科学研究センター, ユニットリーダー (70401164)
• Research Collaborator: IMATAKA hiroaki ; SHIGEMATSU hideki ; YOKOYAMA takeshi ; KASHIWAGI kazuhiro
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2018: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2017: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2016: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: ウイルス / 生体分子 / 電子顕微鏡 / 翻訳 / 翻訳開始

Outline of Final Research Achievements

We determined the cryo-EM structure of the HCV IRES which binds to the 40S ribosomal subunit of the translating 80S ribosome without impeding the ongoing translation. Single-molecule measurements by fluorescence microscopy confirmed that the HCV IRES binds to the translating ribosome. Additional cryo-EM studies showed that the HCV IRES binds to the 40S subunit of the translating 80S ribosome even in the case of the HCV IRES-dependent translation. Biochemical experiments showed that the ribosome translating cap-dependently is a better substrate for the HCV IRES than a free ribosome.

Academic Significance and Societal Importance of the Research Achievements

HCV由来のIRESがこれまで想定されていなかった方法で反応中の翻訳装置リボソームを乗っ取っていることを示した点で画期的である。本研究により明らかになった機構は、HCV由来のIRESと同じような構造を持つ他のウイルスにも共通すると考えられる。従って、IRESとリボソームの40Sサブユニットとの相互作用を阻害する薬剤を開発できれば、効果的な抗ウイルス薬になる可能性がある。

Research Products

• [Int'l Joint Research] UC Berkeley(米国)
• [Journal Article] The Translation Inhibitor Rocaglamide Targets a Bimolecular Cavity between eIF4A and Polypurine RNA (2019)
  • Author(s): Iwasaki Shintaro、Iwasaki Wakana、Takahashi Mari、Sakamoto Ayako、Watanabe Chiduru、Shichino Yuichi、Floor Stephen N.、Fujiwara Koichi、Mito Mari、Dodo Kosuke、Sodeoka Mikiko、Imataka Hiroaki、Honma Teruki、Fukuzawa Kaori、Ito Takuhiro、Ingolia Nicholas T.
  • Journal Title: Molecular Cell Volume: 73 Issue: 4 Pages: 738-748
  • DOI: 10.1016/j.molcel.2018.11.026
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Structural basis for eIF2B inhibition in integrated stress response (2019)
  • Author(s): Kashiwagi Kazuhiro、Yokoyama Takeshi、Nishimoto Madoka、Takahashi Mari、Sakamoto Ayako、Yonemochi Mayumi、Shirouzu Mikako、Ito Takuhiro
  • Journal Title: Science Volume: 364 Issue: 6439 Pages: 495-499
  • DOI: 10.1126/science.aaw4104
  • Peer Reviewed
• [Journal Article] HCV IRES captures an actively translating 80S ribosome (2019)
  • Author(s): Yokoyama Takeshi、Machida Kodai、Iwasaki Wakana、Shigeta Tomoaki、Nishimoto Madoka、Takahashi Mari、Sakamoto Ayako、Yonemochi Mayumi、Harada Yoshie、Shigematsu Hideki、Shirouzu Mikako、Tadakuma Hisashi、Imataka Hiroaki、Ito Takuhiro
  • Journal Title: Molecular Cell Volume: 印刷中 Issue: 6 Pages: 1205-1214
  • DOI: 10.1016/j.molcel.2019.04.022
  • Peer Reviewed / Open Access
• [Journal Article] Dynamic interaction of poly(A)-binding protein with the ribosome (2018)
  • Author(s): Machida Kodai、Shigeta Tomoaki、Yamamoto Yuki、Ito Takuhiro、Svitkin Yuri、Sonenberg Nahum、Imataka Hiroaki
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 17435-17435
  • DOI: 10.1038/s41598-018-35753-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cell-free synthesis of functional antibody fragments to provide a structural basis for antibody-antigen interaction (2018)
  • Author(s): Matsuda, T.
  • Journal Title: PLoS One Volume: 13 Issue: 2 Pages: e0193158-e0193158
  • DOI: 10.1371/journal.pone.0193158
  • Peer Reviewed / Open Access
• [Journal Article] Three-dimensional Structure of eIF2B: A Clue to Understanding the Pathogenesis of CACH/VWM Disease (2017)
  • Author(s): 柏木 一宏
  • Journal Title: BRAIN and NERVE Volume: 69 Issue: 1 Pages: 45-50
  • DOI: 10.11477/mf.1416200635
  • ISSN: 1344-8129, 1881-6096
  • Year and Date: 2017-01-01
• [Journal Article] Solution structure of the first RNA recognition motif domain of human spliceosomal protein SF3b49 and its mode of interaction with a SF3b145 fragment (2017)
  • Author(s): Kuwasako, K.
  • Journal Title: Protein Science Volume: 26 Issue: 2 Pages: 280-290
  • DOI: 10.1002/pro.3080
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Trm5 and TrmD: two enzymes from distinct origins catalyze the identical tRNA modification, m1G37 (2017)
  • Author(s): Goto-Ito, S.
  • Journal Title: Biomolecules Volume: 7 Issue: 1 Pages: 32-32
  • DOI: 10.3390/biom7010032
  • Peer Reviewed / Open Access
• [Journal Article] Crystal structure of eIF2B and insights into eIF2-eIF2B interactions (2017)
  • Author(s): Kashiwagi, K.
  • Journal Title: FEBS Journal Volume: 284 Issue: 6 Pages: 868-874
  • DOI: 10.1111/febs.13896
  • Peer Reviewed / Open Access
• [Journal Article] Methyl transfer by substrate signaling from a knotted protein fold (2016)
  • Author(s): Christian, T.
  • Journal Title: Nature Structural and Molecular Biology Volume: 23 Issue: 10 Pages: 941-948
  • DOI: 10.1038/nsmb.3282
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Rocaglamide Aによる配列特異的な翻訳阻害の構造基盤 (2018)
  • Author(s): 伊藤拓宏
  • Organizer: 第20回日本RNA学会年会
• [Presentation] C型肝炎IRESは翻訳中80Sリボソームに結合し、宿主の翻訳系をハイジャックする (2018)
  • Author(s): 横山武司
  • Organizer: 第20回日本RNA学会年会
• [Presentation] HCV IRES captures a translating 80S ribosome to hijack host translational machinery (2018)
  • Author(s): Ito Takuhiro
  • Organizer: Proteins; from the Cradle to the Grave
  • Int'l Joint Research
• [Presentation] Structural basis for sequence-specific translation inhibition by Rocaglamide A (2018)
  • Author(s): Ito Takuhiro
  • Organizer: Translational Control
  • Int'l Joint Research
• [Presentation] HCV IRES captures a translating 80S ribosome to hijack host translational machinery (2018)
  • Author(s): Yokoyama Takeshi
  • Organizer: Translational Control
  • Int'l Joint Research
• [Presentation] Structural basis for sequence-specific translation repression by eIF4A and Rocaglamide (2017)
  • Author(s): Ito, T.
  • Organizer: ConBio2017
• [Presentation] Crystal structure of eukaryotic translation initiation factor 2B (eIF2B) (2016)
  • Author(s): Kazuhiro Kashiwagi
  • Organizer: The RNA Society / The RNA Society of Japan
  • Place of Presentation: 京都国際会議場(京都)
  • Year and Date: 2016-09-26
  • Int'l Joint Research
• [Presentation] Crystal sturucture of eukaryotic translation initiation factor 2B (2016)
  • Author(s): Takuhiro Ito
  • Organizer: Nascent Chain Biology Meeting 2016
  • Place of Presentation: 富士レークホテル（山梨）
  • Year and Date: 2016-09-01
  • Int'l Joint Research / Invited
• [Presentation] 翻訳開始因子 eIF2B の結晶構造 (2016)
  • Author(s): 柏木 一宏
  • Organizer: 日本蛋白質科学会
  • Place of Presentation: 福岡国際会議場（福岡）
  • Year and Date: 2016-06-08