Synthesis of modified histones and its use for preparation of asymmetric nucleosomes

• Project/Area Number: 18K05316
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: Osaka University
• Principal Investigator: Kawakami Toru 大阪大学, 蛋白質研究所, 准教授 (70273711)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 蛋白質 / ライゲーション / ヒストン / 翻訳後修飾 / ユビキチン / DNAメチル化 / ヌクレオソーム

Outline of Final Research Achievements

Based on the peptide ligation method, we decided to chemically synthesize four types of histones, including post-translational modifications, with the aim of constructing nucleosomes containing selectively modified histones. We also attempt to develop a more efficient ligation method. As a result, we synthesized three types of histones including post-translational modifications, including the results so far. In particular, we succeeded in synthesizing histones containing ubiquitin modification, which is a small protein, and found that the modification activates DNA methyltransferase. Although the synthesis of one type of histone could not be started, it can be synthesized by the similar strategy. We have also found a ligation method with high reaction efficiency and are studying its application.

Academic Significance and Societal Importance of the Research Achievements

遺伝子（DNA）は4種類のヒストンタンパク質からなる8量体に巻き付きヌクレオソームを構成する．これが連なった状態で細胞核に収納される．ヒストンやその関連タンパク質は種々の翻訳後修飾（化学修飾）を受けることで，構造変化やタンパク質間の相互作用を調整して，この遺伝子の発現制御を行っている．これらの修飾は非常に複雑であり，その機能は必ずしも解明されていない．これらの翻訳後修飾を受けたヒストンを選択的に化学合成できるようになれば，それらの機能の解明に非常に有用である．

Research Products

• [Journal Article] Two distinct modes of DNMT1 recruitment ensure stable maintenance DNA methylation (2020)
  • Author(s): A. Nishiyama, C. Mulholland, S. Bultmann, S. Kori, A. Endo, Y. Saeki, W. Qin, C. Trummer, Y. Chiba, H. Yokoyama, S. Kumamoto, T. Kawakami, H. Hojo, G. Nagae, H. Aburatani, K. Tanaka, K. Arita, H. Leonhardt, M. Nakanishi
  • Journal Title: Nat. Commun. Volume: 11 Issue: 1 Pages: 1222-1222
  • DOI: 10.1038/s41467-020-15006-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Enhanced processivity of Dnmt1 by monoubiquitinated histone H3 (2020)
  • Author(s): Y. Mishima, L. Brueckner, S. Takahashi, T. Kawakami, J. Otani, A. Shinohara, K. Takeshita, R. G. Garvilles, M. Watanabe, N. Sakai, H. Takeshima, C. Nachtegael, A. Nishiyama, M. Nakanishi, K. Arita, K. Nakashima, H. Hojo, I. Suetake
  • Journal Title: Genes Cells Volume: 25 Issue: 1 Pages: 22-32
  • DOI: 10.1111/gtc.12732
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chemical synthesis of the ubiquitinated form of histone H3 and its effect on DNA methyltransferase 1 (2019)
  • Author(s): T. Kawakami, Y. Mishima, M. Takazawa, H. Hojo, I. Suetake
  • Journal Title: J. Pept. Sci. Volume: 25 Issue: 9
  • DOI: 10.1002/psc.3200
  • Peer Reviewed
• [Journal Article] Glycopeptide synthesis based on a TFA-labile protection strategy and one-pot four-segment ligation for the synthesis of O-glycosylated histone H2A (2019)
  • Author(s): Y. Asahina, T. Kawakami, H. Hojo
  • Journal Title: Eur. J. Org. Chem. Volume: - Issue: 9 Pages: 1915-1920
  • DOI: 10.1002/ejoc.201801885
  • Peer Reviewed
• [Presentation] Synthesis of Ubiquitinated Histone H3 and its Effect on DNA Methyltransferase 1 (2021)
  • Author(s): Toru Kawakami
  • Organizer: The 18th Akabori Conference
  • Int'l Joint Research
• [Presentation] ライゲーション法を基盤とする翻訳後修飾タンパク質の化学合成 (2021)
  • Author(s): 川上 徹
  • Organizer: 蛋白質研究所セミナー「多角的な視点による蛋白質修飾の機能解明」
• [Presentation] Investigation on stereochemistry of cysteinyl prolyl ester (CPE) peptide for the formation of thioester (2020)
  • Author(s): Eri Sasakura, Hironobu Hojo, Toru Kawakami
  • Organizer: 第57回ペプチド討論会
• [Presentation] ユビキチン化ヒストンH3の化学合成 (2019)
  • Author(s): 川上 徹，高澤雅也，三島優一，北條裕信，末武 勲
  • 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
• [Presentation] 修飾ヒストンの化学合成 (2019)
  • Author(s): 川上 徹
  • Organizer: 蛋白質研究所セミナー「化学によるタンパク質修飾の機能解明を目指して」
  • Invited
• [Presentation] ユビキチン化ヒストンH3の化学合成とその効果 (2019)
  • Author(s): 川上 徹，三島優一，高澤雅也，北條裕信，末武 勲
  • Organizer: 第13回バイオ関連化学シンポジウム
• [Presentation] Chemical Synthesis of Ubiquitinated Histone H3 and Its Effect on DNA Methyltransferase 1 (2019)
  • Author(s): T. Kawakami, Y. Mishima, M. Takazawa, H. Hojo, I. Suetake
  • Organizer: 第56回ペプチド討論会
• [Presentation] 修飾ヒストンの化学合成とその応用例 (2019)
  • Author(s): 川上 徹
  • Organizer: 第42回日本分子生物学会年会
  • Invited
• [Presentation] Chemical synthesis of ubiquitinated histone H3 (2018)
  • Author(s): Toru Kawakami, Masaya Takazawa, Yuichi Mishima, Hironobu Hojo, Isao Suetake
  • Organizer: The 10th international Peptide Symposium
  • Int'l Joint Research
• [Presentation] Synthesis of Ubiquitinated Histone (2018)
  • Author(s): Toru Kawakami
  • Organizer: The 17th Akabori Conference
  • Int'l Joint Research
• [Presentation] A preparation of isopeptide mimetics by the peptide ligation using a thiirane linker (2018)
  • Author(s): Toru Kawakami, Yuichi Mishima, Hironobu Hojo, Isao Suetake
  • Organizer: 35th European Peptide Symposium
  • Int'l Joint Research
• [Remarks]
  • URL: http://www.protein.osaka-u.ac.jp/organic/publication/kawa.html