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Structural biology on the interaction between archaeal CMG helicase and DNA polymerase D

Research Project

Project/Area Number 18K06081
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43020:Structural biochemistry-related
Research InstitutionUniversity of Yamanashi

Principal Investigator

OYAMA Takuji  山梨大学, 大学院総合研究部, 准教授 (60423133)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
KeywordsDNA複製 / CMGヘリカーゼ / 超好熱古細菌 / 結晶構造解析 / CMGヘリカーゼ複合体 / X線結晶構造解析 / DNAポリメラーゼD / X線結晶構造解析 / 古細菌
Outline of Final Research Achievements

In the early stage of DNA replication, the CMG complex, comprising a MCM helicase core (hexamer) and two activators GINS (tetramer) and Cdc45 (GAN in archaea), unwinds the template DNA. Archaea have a DNA replication mechanism similar to that of eukaryotes, and we have recently found a novel interaction between (1) CMG, which is shared among archaea and eukaryotes, and (2) DNA polymerase D, which is unique to archaea. In order to elucidate the functional importance of the new complex interaction revealed by biochemical analysis, we performed X-ray crystallography of a partial complex containing the functional interaction. We succeeded in reconstructing and determining the crystal structure of the complex, elucidated the details of the interaction at the amino acid level, and confirmed the importance of the interaction by biochemical analysis.

Academic Significance and Societal Importance of the Research Achievements

古細菌が持つDNA複製に関わるタンパク質の構造と機能は、真核生物由来のものに似ており、複合体はしばしば簡略化されているため、ヒトを含む真核生物の複雑なシステムを理解するための有用なモデルである。超好熱古細菌由来タンパク質は熱安定性に優れ、構造解析に有利である。一方、古細菌は他生物界と異なる独自システムも併せ持ち、構造生物学的に興味深い。本研究でターゲットとしたDNAポリメラーゼDは古細菌固有酵素であるが、古細菌としてのユニークな特性に加え、構造機能解析を通して真核生物システムとの類似性を見出せれば、学術的に意義高く、生物種間の進化上の関連まで視野を広げれば、知の蓄積の観点で社会的意義に富む。

Report

(5 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Products Report
  • Research Products

    (12 results)

All 2022 2021 2020 2019 2018 Other

All Journal Article (5 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 5 results,  Open Access: 5 results) Presentation (2 results) Remarks (5 results)

  • [Journal Article] Family D DNA polymerase interacts with GINS to promote CMG-helicase in the archaeal replisome.2022

    • Author(s)
      Oki, K., Nagata, M., Yamagami, T., Numata, T., Ishino, S., Oyama, O., and Ishino, Y.
    • Journal Title

      Nucleic Acids Res.

      Volume: 50 Issue: 7 Pages: 3601-3615

    • DOI

      10.1093/nar/gkab799

    • Related Report
      Products Report
    • Peer Reviewed / Open Access
  • [Journal Article] Preparation of co-crystals of human PPARα-LBD and ligand for high-resolution X-ray crystallography2021

    • Author(s)
      Kamata Shotaro、Oyama Takuji、Ishii Isao
    • Journal Title

      STAR Protocols

      Volume: 2 Issue: 1 Pages: 100364-100364

    • DOI

      10.1016/j.xpro.2021.100364

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] PPARα Ligand-Binding Domain Structures with Endogenous Fatty Acids and Fibrates2020

    • Author(s)
      Kamata Shotaro、Oyama Takuji、Saito Kenta、Honda Akihiro、Yamamoto Yume、Suda Keisuke、Ishikawa Ryo、Itoh Toshimasa、Watanabe Yasuo、Shibata Takahiro、Uchida Koji、Suematsu Makoto、Ishii Isao
    • Journal Title

      iScience

      Volume: 23 Issue: 11 Pages: 101727-101727

    • DOI

      10.1016/j.isci.2020.101727

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A crystallization and preliminary X-ray diffraction study of the Arabidopsis thaliana proliferating cell nuclear antigen (PCNA2) alone and in a complex with a PIP-box peptide from Flap endonuclease 12020

    • Author(s)
      Kowalska Ewa、Strza?ka Wojciech、Oyama Takuji
    • Journal Title

      Acta Biochimica Polonica

      Volume: 67 Pages: 49-52

    • DOI

      10.18388/abp.2020_2896

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Direct visualization of DNA baton pass between replication factors bound to PCNA2018

    • Author(s)
      Mayanagi Kouta、Ishino Sonoko、Shirai Tsuyoshi、Oyama Takuji、Kiyonari Shinichi、Kohda Daisuke、Morikawa Kosuke、Ishino Yoshizumi
    • Journal Title

      Scientific Reports

      Volume: 8 Issue: 1 Pages: 16209-16209

    • DOI

      10.1038/s41598-018-34176-2

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] ブタンジオール生産能に関わるSerratia marcescens由来ニコチンアミダーゼのX 線結晶構造解析2019

    • Author(s)
      大山 拓次、大槻 隆司
    • Organizer
      日本微生物生態学会 第33回大会ポスター
    • Related Report
      2019 Research-status Report
  • [Presentation] 結晶構造解析より明らかになったブタンジオール生産に重要なSerratia marcescens ニコチンアミダーゼの多量体構造2019

    • Author(s)
      大山 拓次、大槻 隆司
    • Organizer
      日本結晶学会 2019年度年会ポスター
    • Related Report
      2019 Research-status Report
  • [Remarks] 文部科学省「大学における新型コロナウイルス感染症対策の好事例について」授業事例掲載

    • URL

      https://www.yamanashi.ac.jp/27474

    • Related Report
      2020 Annual Research Report
  • [Remarks] 国立情報学研究所サイバーシンポジウムにて遠隔・対面ハイブリッド講義の事例紹介を発表

    • URL

      https://www.nii.ac.jp/event/other/decs/#16

    • Related Report
      2020 Annual Research Report
  • [Remarks] 研究成果発表(高脂血症治療薬標的分子の構造機能相関に関する論文が国際学術誌iScienceに掲載)

    • URL

      https://www.yamanashi.ac.jp/28446

    • Related Report
      2020 Annual Research Report
  • [Remarks] 日本化学連合シンポジウム「化学研究・教育マネジメント」にて新型コロナ禍でのハイブリッド授業の事例紹介

    • URL

      https://catsj.jp/wp/wp-content/uploads/2020/11/14th_JUCST_sympo.pdf

    • Related Report
      2020 Annual Research Report
  • [Remarks] プレスリリース : アーキア(古細菌)のDNA合成酵素が鋳型DNAの二本鎖を解く機能に貢献! ~アーキア(古細菌)がどのようにDNAを複製するのかを解明~ https://www.yamanashi.ac.jp/wp-content/uploads/2021/09/20210929pr.pdf 2021年9月29日発表

    • Related Report
      Products Report

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Published: 2018-04-23   Modified: 2024-03-28  

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