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Investigation of the regulation mechanism of the replication fork by phosphorylation and intramolecular interaction of Claspin

Research Project

Project/Area Number 18K06124
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43030:Functional biochemistry-related
Research InstitutionTokyo Metropolitan Institute of Medical Science

Principal Investigator

ZHIYING You  公益財団法人東京都医学総合研究所, 基礎医科学研究分野, 主席研究員 (90332270)

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: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
KeywordsClaspin / DNA複製 / フォーク安定化 / DNA結合 / タンパク質のリン酸化 / 分子内結合 / DNA replication / DNA-binding / phosphorylation / de-phosphorylation / MCM helicase / MCM2-7 / Cdc7キナーゼ / DNAヘリカーゼ / 複製フォーク / リン酸化 / MCM
Outline of Final Research Achievements

Claspin is a conserved replication checkpoint mediator and plays a role also in the normal replication fork progression. We reported that Claspin has an acidic patch domain that interacts specifically with Cdc7, and that this domain may also be involved in intramolecular interaction with its N-terminal segment. Purified Claspin from cells barely bound to a fork DNA, whereas the dephosphorylated Claspin changed it to an efficient binder. Claspin form a complex with MCM and inhibit the helicase activity of MCM. In addition, phosphorylation inhibited the DNA binding of Claspin and also inhibited the interaction between the N-terminal and C-terminal of Claspin, suggesting that the phosphorylation of the N-terminal segment regulated the intramolecular interaction and DNA binding of Claspin. We are now identifying the phosphorylated residues that regulate the interactions, and responsible kinase, and their physiological significance.

Academic Significance and Societal Importance of the Research Achievements

外的内的な要因による複製フォークの停止は、ゲノムの不安定化をもたらし、疾患や腫瘍形成の根源的な原因となる。Claspinは、複製ストレス仲介分子としての解析はされてきたが、複製開始進行における役割についての解析はほとんどなされていない。従って、本研究は、複製フォークの制御機構に関して独創的な研究成果をもたらすことで、ガン治療法につながると考えている。今回の研究から、ClaspinがMCMヘリカーゼ活性を抑制することを見出し、リン酸化がClaspinの分子内相互作用およびDNA結合を調節していることにより、複製フォークの安定化、ゲノムの崩壊を防ぐという重要な働きをしている可能性が示唆された。

Report

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

    (4 results)

All 2020 2019 2018

All Presentation (4 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results)

  • [Presentation] DNA複製制御におけるリン酸化によるClaspinの分子内相互作用の調節2020

    • Author(s)
      ○Zhiying You 1, Chi-Chun Yang 1, Hisao Masai1
    • Organizer
      第43回日本分子生物学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] DNA複製制御におけるClaspinの分子内相互作用の役割 Roles of the intramoleular interaction of Claspin in regulation of DNA replication2019

    • Author(s)
      Zhiying You 、Chi-Chun Yang 、Hisao Masai
    • Organizer
      第42回日本分子生物学会
    • Related Report
      2019 Research-status Report
  • [Presentation] The intramolecular interaction mechanism of human Claspin activation2018

    • Author(s)
      Zhiying You 、Chi-Chun Yang 、Hisao Masai
    • Organizer
      DNA metabolism, Genomic Stablity & Human Disease Meeting(Cold Spring Harbor, Asia)
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] Biochemistry of Mcm, the central factor for DNA replication: a hetero-hexameric helicase core complex that does not show DNA helicase activity”2018

    • Author(s)
      Zhiying You
    • Organizer
      首都大学東京Bio-Conference2018
    • Related Report
      2018 Research-status Report
    • Invited

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Published: 2018-04-23   Modified: 2022-01-27  

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