Regulation of the Chromosomal Passenger Complex identifies pathology of chromosomal instability in cancers

• Project/Area Number: 18H04034
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Medium-sized Section 50:Oncology and related fields
• Research Institution: Japanese Foundation for Cancer Research
• Principal Investigator: 広田 亨 公益財団法人がん研究会, がん研究所 実験病理部, 部長 (50421368)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Granted (Fiscal Year 2021)
• Budget Amount: ¥43,810,000 (Direct Cost: ¥33,700,000、Indirect Cost: ¥10,110,000); Fiscal Year 2021: ¥11,830,000 (Direct Cost: ¥9,100,000、Indirect Cost: ¥2,730,000); Fiscal Year 2020: ¥13,910,000 (Direct Cost: ¥10,700,000、Indirect Cost: ¥3,210,000); Fiscal Year 2019: ¥8,580,000 (Direct Cost: ¥6,600,000、Indirect Cost: ¥1,980,000); Fiscal Year 2018: ¥9,490,000 (Direct Cost: ¥7,300,000、Indirect Cost: ¥2,190,000)
• Keywords: がん / 染色体不安定性 / Aurora B / セントロメア / Ｍ期キナーゼ / 染色体分配 / 天然編成領域 / 原子間力顕微鏡 / 分裂期キナーゼ / 動原体 / HP1 / タンパク質構造解析 / M期キナーゼ / 構造解析

Outline of Annual Research Achievements

染色体パセンジャー複合体（Chromosomal Passenger Complex; CPC）はセントロメアに局在し染色体の正確な分配を制御している。この機能においてはAurora Bキナーゼ活性が動原体分子をリン酸化することが必要だが、いかにセントロメアから動原体までの距離を超えて活性を及ぼせるのかが不明であった。本研究ではこれまでにin vitroの再構成系で、CPCは動的な構造をとりそのコンフォマーが伸長することを見出し、セントロメアに局在しながらも活性を動原体まで直接届けることができることが示唆されている。これを踏まえて2020年度は、動的構造を作り出す構造学的解析を進め、細胞内で動的構造を見いだし、その意義を検討するための実験系を作成した。

１） Aurora Bの機能において重要な構造変化であるCPCの伸長の制御を明らかにするために、伸長に主に寄与するINCENPのドメイン解析を行った。つまり伸縮を担うと思われるINCENPの分子内で、形の変化に寄与するドメイン、複合体全体のコンフォマーに寄与するドメイン、その部分がHP1の結合によって制御を受ける可能性を明らかにした。

２）CPCの伸長が実際に細胞内で起こるのかを検討するために、FRET反応を利用したプローブを作成し、これと動原体分子のリン酸化状態、及び動原体と微小管との結合状態を同時に解析し、分裂期進行に伴うCPCの構造変換制御を検討した。さらに、in vitro見出されたCPCの構造変換特性の細胞内での意義を検討するために、CPCの伸張をラパマイシンにより条件的に制御する実験系を構築した。

Current Status of Research Progress

3: Progress in research has been slightly delayed.

Reason

コロナ禍で生化学的な実験が制限されたことが影響し、リコンビナントタンパク質を用いる実験計画の遅延は避けられなかった。しかし再現性のある結果が得られ始めているので、研究の速度を上げつつ試験管内でCPC複合体の構造特性を明らかにしいく。

Strategy for Future Research Activity

試験管内でCPC複合体の構造特性の解析から、CPCが自己集合に関与するドメインの存在が見えてきた。複合体全体の動的構造とともに、自己集合活性がCPCのセントロメアへの濃縮するどのように関わっているのかも合わせて検討していきたい。

Research Products

• [Int'l Joint Research] Colorado State University(米国)
• [Int'l Joint Research] Newcastle University(英国)
• [Int'l Joint Research] Colorado State University(米国)
• [Int'l Joint Research] Colorado State University(米国)
• [Int'l Joint Research] European Molecular Biology Organization(ドイツ)
• [Journal Article] Pathogenicity assessment of variants for breast cancer susceptibility genes based on BRCAness of tumor sample (2021)
  • Author(s): Yoshida Reiko、Hagio Taichi、Kaneyasu Tomoko、Gotoh Osamu、Osako Tomo、Tanaka Norio、Amino Sayuri、Yaguchi Noriko、Nakashima Eri、Kitagawa Dai、Ueno Takayuki、Ohno Shinji、Nakajima Takeshi、Nakamura Seigo、Miki Yoshio、Hirota Toru、Takahashi Shunji、Matsuura Masaaki、Noda Tetsuo、Mori Seiichi
  • Journal Title: Cancer Sci. Volume: 112 Pages: 1310-1319
  • DOI: 10.1111/cas.14803
  • Peer Reviewed / Open Access
• [Journal Article] Prolonged mitosis causes separase deregulation and chromosome nondisjunction (2021)
  • Author(s): Shindo Norihisa、Otsuki Makoto、Uchida Kazuhiko S.K.、Hirota Toru
  • Journal Title: Cell Reports Volume: 34 Pages: 108652-108652
  • DOI: 10.1016/j.celrep.2020.108652
  • Peer Reviewed / Open Access
• [Journal Article] Kinase inhibition profiles as a tool to identify kinases for specific phosphorylation sites (2020)
  • Author(s): Watson Nikolaus A.、Cartwright Tyrell N.、Lawless Conor、Camara-Donoso Marcos、Sen Onur、Sako Kosuke、Hirota Toru、Kimura Hiroshi、Higgins Jonathan M. G.
  • Journal Title: Nature Communications Volume: 11 Pages: 1684-1684
  • DOI: 10.1038/s41467-020-15428-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of a novel nucleolar protein complex required for mitotic chromosome segregation through centromeric accumulation of Aurora B (2020)
  • Author(s): Fujimura, A., Hayashi, Y., Kato, K., Kogure, Y., Kameyama, M., Shimamoto, H., Daitoku, H., Fukamizu, A., Hirota, T., and Kimura, K.
  • Journal Title: Nucleic Acids Research Volume: 48 Pages: 6583-6596
  • DOI: 10.1093/nar/gkaa449
  • NAID: 120007147865
  • Peer Reviewed / Open Access
• [Journal Article] BRAF Mutation in Colorectal Cancers: From Prognostic Marker to Targetable Mutation. (2020)
  • Author(s): Nakayama Izuma, Hirota Toru, Shinozaki Eiji
  • Journal Title: Cancers Volume: 12 Pages: 3236-3236
  • DOI: 10.3390/cancers12113236
  • Peer Reviewed / Open Access
• [Journal Article] Folding the genome into mitotic chromosomes (2019)
  • Author(s): Motoko Takahashi, Toru Hirota
  • Journal Title: Current Opinion in Cell Biology Volume: 60 Pages: 19-26
  • DOI: 10.1016/j.ceb.2019.03.005
  • Peer Reviewed
• [Journal Article] Cdkl-mediated DIAPHI phosphorylation maintains metaphase cortical tension and inactivates the spindle assembly checkpoint at anaphase. (2019)
  • Author(s): Nishimura K, Johmura Y, Deguchi K, Jiang Z, Uchida KSK, Suzuki N, Shimada M, Chiba Y, Hirota T, Yoshimura SH, Kono K, Nakanishi M.
  • Journal Title: Nat commun. Volume: 10 Issue: 1 Pages: 1-10
  • DOI: 10.1038/s41467-019-08957-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Enrichment of CLDN18‐ARHGAP fusion gene in gastric cancers in young adults (2019)
  • Author(s): Nakayama Izuma、Shinozaki Eiji、Sakata Seiji、Yamamoto Noriko、Fujisaki Junko、Muramatsu Yusuke、Hirota Toru、Takeuchi Kengo、Takahashi Shunji、Yamaguchi Kensei、Noda Tetsuo
  • Journal Title: Cancer Science Volume: 110 Pages: 1352-1363
  • DOI: 10.1111/cas.13967
  • Peer Reviewed
• [Journal Article] Dynamics of sister chromatids through the cell cycle: Together and apart (2018)
  • Author(s): Takahashi Motoko、Hirota Toru
  • Journal Title: The Journal of Cell Biology Volume: 217 Pages: 1887-1889
  • DOI: 10.1083/jcb.201804091
  • Peer Reviewed / Open Access
• [Journal Article] Nucleolar integrity during interphase supports faithful Cdk1 activation and mitotic entry (2018)
  • Author(s): Hayashi Yuki、Fujimura Akiko、Kato Kazashi、Udagawa Rina、Hirota Toru、Kimura Keiji
  • Journal Title: Science Advances Volume: 4
  • DOI: 10.1126/sciadv.aap7777
  • Peer Reviewed / Open Access
• [Journal Article] <b>Quantitative analyses of the metaphase-to-anaphase transition reveal differential kinetic regulation for securin and cyclin </b><b>B1 </b> (2018)
  • Author(s): Makoto Konishi, Norihisa Shindo, Masataka Komiya, Kozo Tanaka, Takehiko Itoh, and Toru Hirota
  • Journal Title: Biomed. Res. Volume: 39 Issue: 2 Pages: 75-85
  • DOI: 10.2220/biomedres.39.75
  • NAID: 130006708566
  • ISSN: 0388-6107, 1880-313X
  • Peer Reviewed / Open Access
• [Presentation] がん細胞における染色体不安定性の病理機構 (2020)
  • Author(s): 広田 亨
  • Organizer: 第79回日本癌学会学術総会
  • Int'l Joint Research / Invited
• [Presentation] 分裂期染色体制御の破綻によるがん幹細胞のプロイディ変化 (2020)
  • Author(s): 趙 民知、サンペトラ・オルテア、佐谷秀行、広田 亨
  • Organizer: 第79回日本癌学会学術総会
  • Int'l Joint Research
• [Presentation] セパレース制御異常が誘導するがん細胞の染色体分配異常 (2020)
  • Author(s): 進藤軌久、広田 亨
  • Organizer: 第79回日本癌学会学術総会
  • Int'l Joint Research
• [Presentation] 液液相分離を介したAurora Bキナーゼの活性化の時空間的な制御機構 (2020)
  • Author(s): 野澤竜介、広田 亨
  • Organizer: 第79回日本癌学会学術総会
  • Int'l Joint Research
• [Presentation] A robust mitotic regulation facilitates karyotypic evolution in cancer stem cells (2020)
  • Author(s): Toru Hirota
  • Organizer: The 43rdnd Annual meeting of the Molecular Biology Society of Japan
  • Int'l Joint Research / Invited
• [Presentation] Chromosomal passenger complex creates a nuclear compartment regulating Aurora B kinase activity through liquid-liquid phase separation (2020)
  • Author(s): Ryu-Suke Nozawa, Toru Hirota
  • Organizer: The 43rdnd Annual meeting of the Molecular Biology Society of Japan
  • Int'l Joint Research
• [Presentation] The homeostatic control of chromosomes. Symposium (Organizer) (2019)
  • Author(s): Toru Hirota and Katsuhiko Shirahige
  • Organizer: The 71st Annual meeting of the Japan Society for Cell Biology (JSCB)
  • Int'l Joint Research
• [Presentation] A robust control of mitotic transitions prevents chromosome missegregation. (2019)
  • Author(s): Toru Hirota
  • Organizer: International Symposium Cancer Etiology. Tohoku Forum for Creativity
  • Int'l Joint Research / Invited
• [Presentation] Chromosome dynamics in cancers. Symposium (Organizer) (2019)
  • Author(s): Toru Hirota and Satoshi Tashiro
  • Organizer: The 78th Annual meeting of the Japanese Cancer Association (JCA)
  • Int'l Joint Research
• [Presentation] Possible contribution of chromosomal instability in cancer progression. (2019)
  • Author(s): Jo Minji, Oltea Sampetrean, Hideyuki Saya, Toru Hirota
  • Organizer: The 78th Annual meeting of the Japanese Cancer Association (JCA)
  • Int'l Joint Research / Invited
• [Presentation] Spatial control of Aurora B activity at centromeres in mitosis. (2019)
  • Author(s): Toru Hirota
  • Organizer: The 42nd Annual meeting of the Molecular Biology Society of Japan (MBSJ)
  • Int'l Joint Research / Invited
• [Presentation] Characterization of DNA binding mode of human condensin I and II. (2018)
  • Author(s): Motoko Takahashi and Toru Hirota
  • Organizer: The 7th Meeting on Grant-in-Aid for scientific research on innovative areas “Chromosome Orchestration System”
  • Int'l Joint Research
• [Presentation] A robust transition from metaphase to anaphase prevents chromosome missegregation (2018)
  • Author(s): Toru Hirota
  • Organizer: The 77th Annual meeting of the Japanese Cancer Association (JCA)
  • Int'l Joint Research / Invited
• [Presentation] Single stranded DNA in interphase is a key chromatin structure for mitotic chromosome organization (2018)
  • Author(s): Motoko Takahashi and Toru Hirota
  • Organizer: The 11th 3R+3C Symposium
  • Int'l Joint Research
• [Presentation] Troubles of the brake and engine driving chromosome segregation in cancer (2018)
  • Author(s): Toru Hirota
  • Organizer: Symposium. Genoproteomic signature of human diseases
  • Int'l Joint Research / Invited
• [Presentation] A robust control of metaphase-to-anaphase transition ensures stable chromosome segregation (2018)
  • Author(s): Toru Hirota
  • Organizer: The 41st Annual meeting of the Molecular Biology Society of Japan (MBSJ)
  • Int'l Joint Research / Invited
• [Book] 染色体分配：マルチステップに進む姉妹染色分体の分離. 実験医学 (2018)
  • Author(s): 内田和彦、広田 亨
  • Total Pages: 6
  • Publisher: 羊土社
• [Remarks] がん研 広田亨研究室
  • URL: https://www.jfcr.or.jp/tci/exppathol/index.html
• [Remarks] がん研究所 実験病理部
  • URL: https://www.jfcr.or.jp/laboratory/department/experiment/index.html
• [Remarks] がん研究所実験病理部・広田亨研究室
  • URL: http://www.jfcr.or.jp/tci/exppathol/
• [Remarks] Researchmap（広田 亨）
  • URL: https://researchmap.jp/toru_hirota/
• [Remarks] がん研究所実験病理部
  • URL: http://www.jfcr.or.jp/tci/exppathol/
• [Remarks] がん研究会がん研究所
  • URL: http://www.jfcr.or.jp/laboratory/index.html
• [Patent(Industrial Property Rights)] HP1の機能に着目した抗がん剤のスクリーニング方法及び評価系 (2019)
  • Inventor(s): 広田 亨、阿部 優介
  • Industrial Property Rights Holder: 広田 亨、阿部 優介
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-168849
  • Filing Date: 2019
• [Patent(Industrial Property Rights)] HP1の機能に着目した抗がん剤のスクリーニング方法及び評価系 (2019)
  • Inventor(s): 広田 亨、阿部 優介
  • Industrial Property Rights Holder: 広田 亨、阿部 優介
  • Industrial Property Rights Type: 特許
  • Filing Date: 2019
  • Acquisition Date: 2019
• [Funded Workshop] The 37th Chromosome Workshop (2019)
• [Funded Workshop] The 11th 3R+3C Symposium (2018)