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Elucidation of Molecular Sorting Mechanisms using Cryo-electron microscopy

Research Project

Project/Area Number 21H02654
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 48010:Anatomy-related
Research InstitutionUniversity of Yamanashi

Principal Investigator

Oda Toshiyuki  山梨大学, 大学院総合研究部, 教授 (20569090)

Co-Investigator(Kenkyū-buntansha) 柳澤 春明  東京大学, 大学院医学系研究科(医学部), 講師 (70466803)
新森 英之  山梨大学, 大学院総合研究部, 准教授 (40311740)
Project Period (FY) 2021-04-01 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
Fiscal Year 2023: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2022: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2021: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Keywordsクライオ電子顕微鏡 / 繊毛 / ダイニン
Outline of Research at the Start

細胞内には数千種類以上もタンパク質が混在しており、これらの分子が適切な場所で働くことで、生命活動が維持されている。そのために細胞には、タンパク質を識別し、正しいコンパートメントに局在させる「分子ソート機構」が存在する。人体に於いて器官形成や免疫、感覚など重要な生理機能を担っている繊毛はこのような細胞内コンパートメントの好例であり、繊毛の機能維持には繊毛タンパク質を正確にソートすることが必要であると考えられる。本研究では、繊毛をターゲットとした分子ソート機構が働いていると考えられる2つの繊毛基部構造に注目し、繊毛タンパク質が正しく組み上げられ、正しい場所に運ばれる分子メカニズムを解明する。

Outline of Final Research Achievements

Cilia are essential cellular organelles necessary for cell signaling and motility. The central structure of cilia, the axoneme, adopts the so-called '9+2' structure composed of nine peripheral microtubules and two central pair microtubules. However, the molecular mechanisms that construct this complex structure remain largely unknown. This study identifies a molecule at the base of one of the main projections of the central pair microtubules, the C2a structure, and demonstrates that its presence is essential for the construction of the C2a structure. Additionally, by utilizing the incorporation of the antibiotic puromycin into nascent proteins, we have revealed the existence of a precursor pool at the base of cilia. Furthermore, by genetically manipulating the carboxyl-terminal ends of tubulin, we have shed light on some aspects of the construction mechanism of the central pair microtubules.

Academic Significance and Societal Importance of the Research Achievements

本研究は繊毛を持つ緑藻クラミドモナスを用いて、繊毛の動きと構造に欠かせないタンパク質の働きを明らかにしました。繊毛は呼吸器や生殖系など人体の多くの部位で重要な役割を果たしており、この研究は病気の治療法の開発に貢献する可能性を秘めています。具体的には、抗生物質の特殊な性質を利用した新規分析法を用いて繊毛タンパク質の合成を追跡し、また遺伝子操作により繊毛の微細構造を解明する手法を開発しました。これらの成果は、繊毛異常が引き起こす病気の理解を深め、治療に向けた新たな道を開く基盤となります。

Report

(4 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Annual Research Report
  • 2021 Annual Research Report
  • Research Products

    (14 results)

All 2024 2023 2022 2021 Other

All Int'l Joint Research (1 results) Journal Article (6 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 6 results) Presentation (6 results) (of which Int'l Joint Research: 5 results,  Invited: 4 results) Book (1 results)

  • [Int'l Joint Research] Univ Massachusetts Med Sch(米国)

    • Related Report
      2021 Annual Research Report
  • [Journal Article] Tubulin glycylation controls ciliary motility through modulation of outer-arm dyneins2024

    • Author(s)
      Tomohiro Kubo1, Rinka Sasaki, and Toshiyuki Oda
    • Journal Title

      Molecular Biology of the Cell

      Volume: -

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] α- and β-tubulin C-terminal tails with distinct modifications are crucial for ciliary motility and assembly2023

    • Author(s)
      Kubo Tomohiro、Tani Yuma、Yanagisawa Haru-Aki、Kikkawa Masahide、Oda Toshiyuki
    • Journal Title

      Journal of Cell Science

      Volume: 136 Issue: 16

    • DOI

      10.1242/jcs.261070

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Cryo-EM elucidates the uroplakin complex structure within liquid-crystalline lipids in the porcine urothelial membrane2023

    • Author(s)
      Yanagisawa Haruaki、Kita Yoshihiro、Oda Toshiyuki、Kikkawa Masahide
    • Journal Title

      Communications Biology

      Volume: 6 Issue: 1 Pages: 1018-1018

    • DOI

      10.1038/s42003-023-05393-x

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Cryo-electron tomography of Birbeck granules reveals the molecular mechanism of langerin lattice formation2022

    • Author(s)
      T. Oda, H. Yanagisawa, H. Shinmori, Y. Ogawa, T. Kawamura
    • Journal Title

      eLife

      Volume: 11

    • DOI

      10.7554/elife.79990

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Chlamydomonas FAP70 is a component of the previously uncharacterized ciliary central apparatus projection C2a2021

    • Author(s)
      Hou Yuqing、Zhao Lei、Kubo Tomohiro、Cheng Xi、McNeill Nathan、Oda Toshiyuki、Witman George B.
    • Journal Title

      Journal of Cell Science

      Volume: 134 Issue: 12 Pages: 1-11

    • DOI

      10.1242/jcs.258540

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Neural and behavioral control in Caenorhabditis elegans by a yellow-light?activatable caged compound2021

    • Author(s)
      Takahashi Hironori、Kamiya Mako、Kawatani Minoru、Umezawa Keitaro、Ukita Yoshiaki、Niwa Shinsuke、Oda Toshiyuki、Urano Yasuteru
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 118 Issue: 6

    • DOI

      10.1073/pnas.2009634118

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] Cryo-electron microscopy of uroplakin complex.2023

    • Author(s)
      小田賢幸
    • Organizer
      第79回日本顕微鏡学会学術講演会
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Cryo-electron tomography of Birbeck granules reveals the molecular mechanism of langerin lattice formation2023

    • Author(s)
      小田賢幸
    • Organizer
      第128回日本解剖学会全国学術集会
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Cryo-electron tomography of Birbeck granules2022

    • Author(s)
      小田賢幸
    • Organizer
      第78回日本顕微鏡学会学術講演会
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Cryo-electron tomography of Birbeck granules2022

    • Author(s)
      小田賢幸
    • Organizer
      第82回日本解剖学会中部支部学術集会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Cryo-electron tomography of Birbeck granules.2022

    • Author(s)
      Toshiyuki Oda
    • Organizer
      The 78th Annual Meeting of the Japanese Society of Microscopy
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Cryo-electron tomography of cardiac myofibrils reveals a 3D lattice spring within the Z-discs2021

    • Author(s)
      Toshiyuki Oda
    • Organizer
      The 77th Annual Meeting of the Japanese Society of Microscopy
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Book] クライオ電子顕微鏡ハンドブック2023

    • Author(s)
      難波 啓一ら
    • Total Pages
      460
    • Publisher
      (株)エヌ・ティー・エス
    • ISBN
      9784860438043
    • Related Report
      2023 Annual Research Report

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Published: 2021-04-28   Modified: 2025-01-30  

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