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Structural analysis of large protein complexes based on the novel evolutionary indicator

Research Project

Project/Area Number 17K07321
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Structural biochemistry
Research InstitutionHigh Energy Accelerator Research Organization

Principal Investigator

ADACHI NARUHIKO  大学共同利用機関法人高エネルギー加速器研究機構, 物質構造科学研究所, 特任准教授 (70707489)

Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords分子進化 / 構造生物学 / 転写 / 複合体 / 蛋白質 / 遺伝子
Outline of Final Research Achievements

The general transcription factor TFIID is involved in the regulation of 90% of the expression of all genes and plays a central role in transcriptional initiation. However, since it is a complex of 15 subunits with a molecular weight of 1 MDa, its high-resolution three-dimensional structure has not yet been analyzed. We have established a large scale purification method for TFIID and its five interacting factors. The next problem is the mixing order of factors. Based on a new index of molecular evolution developed recently, we have come up with the idea of mixing factors in the order in which they were born during evolution. In this study, we performed A. Crystallization and crystal structure analysis of TFIID, B. Molecular evolutionary analysis, and C. Reconstruction and three-dimensional structure analysis of TFIID-containing complexes.

Academic Significance and Societal Importance of the Research Achievements

生体内において、タンパク質などの生体分子は複合体を形成して働いている。したがって、生体分子の機能解明には、複合体の状態で立体構造を解析することが重要である。しかし、複合体の立体構造解析を行うためには、溶液中で複合体を安定に維持する条件と、複合体を結晶化する(もしくは良好なクライオグリッドを作製する)条件を一致させる必要があるため、一般的に、複合体の立体構造解析は極めて困難であることが知られている。今回の研究は、現代の生命科学において必要不可欠であるものの、いまだに困難とされる複合体の立体構造解析に対して、一般的な解を与えるといった学術的な意義を有する。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (28 results)

All 2020 2019 2018 2017 Other

All Int'l Joint Research (2 results) Journal Article (3 results) (of which Open Access: 3 results,  Peer Reviewed: 2 results) Presentation (19 results) (of which Int'l Joint Research: 7 results,  Invited: 11 results) Book (1 results) Remarks (3 results)

  • [Int'l Joint Research] Harvard University(米国)

    • Related Report
      2019 Annual Research Report
  • [Int'l Joint Research] 上海科学技術大学(中国)

    • Related Report
      2019 Annual Research Report
  • [Journal Article] Crystallographic and cryogenic electron microscopic structures and enzymatic characterization of sulfur oxygenase reductase from Sulfurisphaera tokodaii2020

    • Author(s)
      Yuta Sato, Takashi Yabuki, Naruhiko Adachi, Toshio Moriya, Takatoshi Arakawa, Masato Kawasaki, Chihaya Yamada, Toshiya Senda, Shinya Fushinobu, Takayoshi Wakagi
    • Journal Title

      bioRxiv

      Volume: -

    • DOI

      10.1101/2020.05.03.074773

    • Related Report
      2019 Annual Research Report
    • Open Access
  • [Journal Article] Crystal structure of the DNA-binding domain of the LysR-type transcriptional regulator CbnR in complex with a DNA fragment of the recognition-binding site in the promoter region.2018

    • Author(s)
      Koentjoro MP, Adachi N, Senda M, Ogawa N, Senda T.
    • Journal Title

      FEBS J.

      Volume: 285(5) Issue: 5 Pages: 977-989

    • DOI

      10.1111/febs.14380

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Leading role of TBP in the Establishment of Complexity in Eukaryotic Transcription Initiation Systems2017

    • Author(s)
      Kawakami Eiryo、Adachi Naruhiko、Senda Toshiya、Horikoshi Masami
    • Journal Title

      Cell Reports

      Volume: 21 Issue: 13 Pages: 3941-3956

    • DOI

      10.1016/j.celrep.2017.12.034

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] クライオ電子顕微鏡:気になっていること・困っていること@KEK2019

    • Author(s)
      安達成彦
    • Organizer
      日本顕微鏡学会・第75回学術講演会(口頭発表)
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Structure analysis of transcription related complexes and installation of cryo-EM in KEK2019

    • Author(s)
      安達成彦
    • Organizer
      第19回日本蛋白質科学会年会・第71回日本細胞生物学会大会 合同年次大会(口頭発表)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] クライオ電子顕微鏡で生体分子の形を明らかにする2019

    • Author(s)
      安達成彦
    • Organizer
      KEK公開講座「生物学におけるクライオ電子顕微鏡」(口頭発表)
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Structure analysis of transcription related complexes and installation of cryo-EM in KEK2019

    • Author(s)
      Naruhiko Adachi, Masato Kawasaki, Toshio Moriya, Fumiaki Yumoto, Kotaro Koiwai, Akira Shinoda, Yusuke Yamada, Toshiya Senda
    • Organizer
      The 2019 American Crystallographic Association Annual Meeting(ポスター発表)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 単粒子解析の原理と実際:KEKにおける共同利用型クライオ電子顕微鏡の立ち上げを例として2019

    • Author(s)
      安達成彦
    • Organizer
      日本プロテオーム学会2019年大会・第70回日本電気泳動学会総会(口頭発表)
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Structure analysis of transcription related complexes and installation of cryo-EM in KEK2019

    • Author(s)
      Naruhiko Adachi, Masato Kawasaki, Toshio Moriya, Fumiaki Yumoto, Kotaro Koiwai, Akira Shinoda, Yusuke Yamada, Toshiya Senda
    • Organizer
      Cold Spring Harbor Laboratory meeting, Mechanisms of eukaryotic transcription(ポスター発表)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] KEKクライオ電顕の現状2019

    • Author(s)
      千田俊哉・安達成彦・川崎政人・守屋俊夫・山田悠介・篠田晃・小祝孝太郎
    • Organizer
      生理研研究会「クライオ電子顕微鏡によるタンパク質の高分解能単粒子構造解析」
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] KEKにおけるクライオ電顕施設の紹介2019

    • Author(s)
      千田俊哉・安達成彦・川崎政人・守屋俊夫・山田悠介・篠田晃・小祝孝太郎
    • Organizer
      第42回日本分子生物学会年会(口頭発表)
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] マルチサブユニット複合体の研究とKEKクライオ電子顕微鏡の現状2019

    • Author(s)
      安達成彦
    • Organizer
      蛋白研セミナー(第1回構造生命科学研究会)
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] 遺伝子発現制御に関わるマルチサブユニット複合体の構造・機能・進化2019

    • Author(s)
      安達成彦
    • Organizer
      第1回ゲノム・進化・構造の会
    • Related Report
      2018 Research-status Report
  • [Presentation] 試料調製、測定、解析で困ったこととその解決方法の紹介@KEK2019

    • Author(s)
      安達成彦
    • Organizer
      第1回クライオEMネットワーク・ユーザーグループミーティング
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] KEKにおけるクライオ電顕施設の立ち上げと共同利用2019

    • Author(s)
      安達成彦
    • Organizer
      第5回筑波大-KEK連携セミナー
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] 遺伝子発現制御に関わるタンパク質巨大複合体の構造・機能・進化2018

    • Author(s)
      安達成彦
    • Organizer
      第3回CUBSSミーティング
    • Related Report
      2018 Research-status Report
  • [Presentation] Leading role of TBP in the establishment of complexity in eukaryotic transcription initiation systems2018

    • Author(s)
      Naruhiko Adachi
    • Organizer
      The 18th Annual meeting of PSSJ
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] Leading role of TBP in the establishment of complexity in eukaryotic transcription initiation systems2018

    • Author(s)
      Naruhiko Adachi, Eiryo Kawakami, Toshiya Senda, Masami Horikoshi
    • Organizer
      The 32th Annual meeting of Protein Society
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] KEKのクライオ電子顕微鏡の現状2018

    • Author(s)
      安達成彦
    • Organizer
      BINDS測定技術交流会
    • Related Report
      2018 Research-status Report
  • [Presentation] タンパク質複合体サンプルの調製とスクリーニング2018

    • Author(s)
      安達成彦
    • Organizer
      PF研究会
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] Leading role of TBP in the establishment of complexity in eukaryotic transcription initiation systems2018

    • Author(s)
      Naruhiko Adachi, Eiryo Kawakami, Toshiya Senda, Masami Horikoshi
    • Organizer
      Cold Spring Harbor meeting
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Improved method for soluble expression and rapid purification of yeast TFIIA2017

    • Author(s)
      Naruhiko Adachi, Kyohei Aizawa, Yuka Kratzer, Shinya Saijo, Nobutaka Shimizu, Toshiya Senda.
    • Organizer
      Cold Spring Harbor Lab Meeting: MECHANISMS OF EUKARYOTIC TRANSCRIPTION
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Book] 実験医学増刊・イメージング時代の構造生命科学2020

    • Author(s)
      安達成彦、千田俊哉
    • Total Pages
      6
    • Publisher
      羊土社
    • ISBN
      9784758103855
    • Related Report
      2019 Annual Research Report
  • [Remarks] 高エネルギー加速器研究機構・物質構造科学研究所・構造生物学研究センター

    • URL

      https://www2.kek.jp/imss/sbrc/

    • Related Report
      2019 Annual Research Report
  • [Remarks] KEK・IMSS・構造生物学研究センター(SBRC)

    • URL

      https://www2.kek.jp/imss/sbrc/

    • Related Report
      2018 Research-status Report
  • [Remarks] 高エネルギー加速器研究機構・物質構造科学研究所・構造生物学研究センター

    • URL

      http://www2.kek.jp/imss/sbrc/

    • Related Report
      2017 Research-status Report

URL: 

Published: 2017-04-28   Modified: 2021-02-19  

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