A new method to evaluate the function of mitotic kinases by monitoring chromosome dynamics

• Project/Area Number: 23651243
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Chemical biology
• Research Institution: Japanese Foundation For Cancer Research
• Principal Investigator: HIROTA Toru 公益財団法人がん研究会, がん研究所実験病理部, 部長 (50421368)
• Co-Investigator(Renkei-kenkyūsha): ABE Yusuke 公益財団法人がん研究会, がん研究所・実験病理部, 博士研究員 (50615896) ; UCHIDA Kazuhiko 公益財団法人がん研究会, がん研究所・実験病理部, 博士研究員 (40380555) ; SHINDO Norihisa 公益財団法人がん研究会, がん研究所・実験病理部, 博士研究員 (00512253) ; KUMADA Kazuki 公益財団法人がん研究会, がん研究所・実験病理部, 博士研究員 (10370149)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2012: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: M期 / 染色体 / がん細胞 / キナーゼ阻害薬 / 細胞分裂 / 染色体動態 / がん / 分裂期キナーゼ / 異数性 / Aurora B / Plk1

Research Abstract

Mitotic kinases play crucial roles in separating chromosomes correctly and safely as cells divide. Dysfunction of mitotic kinases therefore causes defective chromosome movements and segregation. In this study, we aimed to develop an assay to indicate cryptic dysfunction of mitotic kinases by monitoring chromosome behavior during mitosis, which is otherwise difficult to detect in conventional means. We found that increasing dose of specific inhibitors of mitotic kinases, caused abnormal chromosomal movements progressively. This what we called the stepwise-challenging assay was then used to address the function of mitotic kinases in several cancer cells that are known to have chromosomal instability

Research Products

• [Journal Article] The nucleoporin Nup188 is required for chromosome alignment in mitosis (2013)
  • Author(s): 伊藤剛、杉野史朗、池田真教、水口万裕美、菅野新一郎、M.A.Amin、家村顕自、安井明、広田亨、田中耕三
  • Journal Title: Cancer Science Volume: (in press) Issue: 7 Pages: 871-879
  • DOI: 10.1111/cas.12159
  • Peer Reviewed
• [Journal Article] Separase sensor reveals dual roles for separase coordinating cohesin cleavage and cdk1 inhibition (2012)
  • Author(s): Shindo, N., Kumada, K., *Hirota, T
  • Journal Title: Dev. Cell Volume: 23 Pages: 112-123
• [Journal Article] HDAC8 mutations in Cornelia de Lange syndrome affect the cohesin acetylation cycle (2012)
  • Author(s): Deardorff MA., Bando, M., Nakato, R., Watrin, E., Itoh, T., Minamino, M.,Saitoh,K., Komata, M., Katou, Y., Clark, D., Cole, KE., De Baere, E., Decroos, C., Di Donato, N., Ernst, S., Francey, LJ., Gyftodimou, Y., Hirashima, K., Hullings, M., Ishikawa, Y., Jaulin, C., Kaur, M., Kiyono, T., Lombardi, PM., Magnaghi-Jaulin, L., Mortier, GR., Nozaki, N., Petersen, MB., Seimiya, H., Siu, VM., Suzuki, Y., Takagaki, K., Wilde, JJ., Willems, PJ, Prigent, C., Gillessen-Kaesbach, G., Christianson, DW., Kaiser, FJ., Jackson, LG., Hirota, T., Krantz ID., *Shirahige K
  • Journal Title: Nature Volume: 489 Pages: 313-317
• [Journal Article] Separase sensor reveals dual roles for separase coordinating cohesin cleavage and cdk1 inhibition. (2012)
  • Author(s): Shindo, N.
  • Journal Title: Developmental Cell Volume: 23 Issue: 1 Pages: 112-123
  • DOI: 10.1016/j.devcel.2012.06.015
  • Peer Reviewed
• [Journal Article] HDAC8 mutations in Cornelia de Lange syndrome affect the cohesin acetylation cycle. (2012)
  • Author(s): Deardorff MA
  • Journal Title: Nature Volume: 489(7415) Issue: 7415 Pages: 313-7
  • DOI: 10.1038/nature11316
  • Peer Reviewed
• [Journal Article] The initial phase of chromosome condensation requires Cdk1-mediated phosphorylation of the CAP-D3 subunit of condensin II (2011)
  • Author(s): Abe, S., Nagasaka, K., Hirayama, Y., Kozuka-Hata, H., Oyama, M., Aoyagi, Y., Obuse, C., and *Hirota, T
  • Journal Title: Genes Dev Volume: 25 Pages: 863-874
• [Journal Article] CAMP (C13orf8, ZNF828) is a novel regulator of kinetochore-microtubule attachment (2011)
  • Author(s): Ito, G., Kanno, S., Uchida, K.S.K, Chiba, S., Sugino, S., Watanabe, K., Mizuno, K., Yasui, A., Hirota, T., and *Tanaka, K
  • Journal Title: EMBO J Volume: 30 Pages: 130-144
• [Journal Article] Differential control of Eg5 dependent centrosome separation by Plk1 and Cdk1 (2011)
  • Author(s): Smith, E., Hegarat, N., Vesely, C., Roseboom, I., Streicher, H., Straatman, K., Flynn, H., Skehel, M., Hirota, T., Kuriyama, R., *Hochegger, H
  • Journal Title: EMBO J Volume: 30 Pages: 2233-2245
• [Journal Article] The initial phase of chromosome condensation requires Cdk1-mediated phosphorylation of the CAP-D3 subunit of condensin II. (2011)
  • Author(s): Abe, S., Nagasaka, K., Hirayama, Y., Kozuka-Hata, H., Oyama, M., Aoyagi, Y., Obuse, C., Hirota, T.
  • Journal Title: Genes & Dev. Volume: 25 Pages: 863-874
  • Peer Reviewed
• [Journal Article] Differential control of Eg5 dependent controsome separation by Plk1 and Cdk1. (2011)
  • Author(s): Smith, E., Hegarat, N., Vesely, C., Roseboom, I., Larch,C., Streicher, H., Straataman, K., Flynn, H., Skehel, M., Hirota, T., Kuriyama, R., Hochegger, H.
  • Journal Title: EMBO J. Volume: 30 Pages: 2233-2245
  • Peer Reviewed
• [Journal Article] CAMP (C13orf8, ZNF828) is a novel regulator of kinetochore-microtubule attachment. (2011)
  • Author(s): Ito, G., Kanno, S., Uchida, KS,, Chiba, S., Sugino, S., Watanabe, K., Mizuno, K., Yasui, A., Hirota, T., Tanaka, K.
  • Journal Title: EMBO J. Volume: 30 Pages: 130-144
  • Peer Reviewed
• [Presentation] How cells ensure separation of sister chromatids in mitosis (2013)
  • Author(s): Hirota, T
  • Organizer: International Symposium on Molecular Medicine
  • Place of Presentation: (Suwon, Korea)
  • Year and Date: 2013-02-28
• [Presentation] M期プロテアーゼ・セパレースの活性プローブによって分かったこと (2013)
  • Author(s): 広田 亨
  • Organizer: 東京理科大学先端生命研・水曜セミナー
  • Place of Presentation: (野田)
  • Year and Date: 2013-01-30
• [Presentation] Consecutive functions of separase ensure the chromosome segregation switch (2012)
  • Author(s): Hirota, T
  • Organizer: Seminar at Research Institute of Genetics and Development
  • Place of Presentation: (Rennes, France)
  • Year and Date: 2012-09-04
• [Presentation] How cells ensure separation of sister chromatids in mitosis (2012)
  • Author(s): Hirota, T
  • Organizer: Seminar at Oxford University Dunn School of Pathology
  • Place of Presentation: (Oxford, UK)
  • Year and Date: 2012-04-19
• [Presentation] How cells ensure separation of sister chromatids in mitosis (2012)
  • Author(s): Hirota, T
  • Organizer: Seminar at Cancer Research UK London Research Insitute
  • Place of Presentation: (London, UK)
  • Year and Date: 2012-04-18
• [Presentation] How cells ensure separation of sister chromatids in mitosis (2012)
  • Author(s): Hirota, T
  • Organizer: Seminar at Imperial College London
  • Place of Presentation: (London, UK)
  • Year and Date: 2012-04-13
• [Presentation] がん細胞における異数性の発生要因を考える (2012)
  • Author(s): 広田 亨
  • Organizer: 第28回新潟県立中央病院集談会・特別講演
  • Place of Presentation: (上越)
  • Year and Date: 2012-03-02
• [Presentation] How cells ensure separation of sister chromatids in mitosis (2012)
  • Author(s): Hirota, T
  • Organizer: Jacques Monad Conference "Cell cycle"
  • Place of Presentation: (Roscoff, France)
• [Presentation] 染色体分離を決行するM期プロテアーゼ・セパレースの活性制御 (2012)
  • Author(s): 広田 亨
  • Organizer: 特定領域研究「細胞増殖制御」終了シンポジウム
  • Place of Presentation: (大岡山)
• [Presentation] How cells ensure trigger chromosome assembly in cells entering mitosis (2012)
  • Author(s): Hirota, T
  • Organizer: Symposium Proteolysis and Cohesion
  • Place of Presentation: (Vienna, Austria)
• [Presentation] Dual functions of separase ensure switch-like initiation of anaphase (2012)
  • Author(s): Hirota, T
  • Organizer: Joint Spring Meeting of BSCB/BSCB/JSDB
  • Place of Presentation: (Coventry, UK)
• [Presentation] アナログ感受性変異体によるGreatwallキナーゼの機能解析 (2012)
  • Author(s): 阿部優介、広田 亨
  • Organizer: 第29回染色体ワークショップ
  • Place of Presentation: (秋保温泉)
• [Presentation] Separase の自己切断による染色体分配制御 (2012)
  • Author(s): 熊田和貴、広田 亨
  • Organizer: 第29回染色体ワークショップ
  • Place of Presentation: (秋保温泉)
• [Presentation] Separase は染色体分離において一人二役を果たす (2012)
  • Author(s): 進藤軌久、広田 亨
  • Organizer: 第29回染色体ワークショップ
  • Place of Presentation: (秋保温泉)
• [Presentation] How cells ensure trigger chromosome assembly in cells entering mitosis. (2012)
  • Author(s): Toru Hirota
  • Organizer: Symposium Proteolysis and Cohesion.
  • Place of Presentation: Vienna, Austria
  • Invited
• [Presentation] How cells ensure separation of sister chromatids in mitosis. (2012)
  • Author(s): Toru Hirota
  • Organizer: Jacques Monad Conference “Cell cycle”
  • Place of Presentation: Roscoff, France
  • Invited
• [Presentation] How mitotic kinases control chromosome segregation (2011)
  • Author(s): Hirota, T
  • Organizer: 熊本大学GCOE・リエゾンラボ研究会
  • Place of Presentation: (熊本)
  • Year and Date: 2011-11-02
• [Presentation] 染色体分離の正確性を保証するメカニズム (2011)
  • Author(s): 広田 亨
  • Organizer: ケミカルバイオロジー勉強会・理化学研究所
  • Place of Presentation: (和光)
  • Year and Date: 2011-10-26
• [Presentation] Heterochromatin protein 1 is an essential component of the chromosomal passenger complex to prevent mis- segregation of chromosomes (2011)
  • Author(s): Hirota, T
  • Organizer: 平成23年度遺伝研研究会「クロマチンダイナミクスの分子機構」
  • Place of Presentation: (三島)
  • Year and Date: 2011-10-20
• [Presentation] The switch-like activation of separase ensures chromosome segregation (2011)
  • Author(s): Hirota, T
  • Organizer: The GCOE International Workshop "Mitosis"
  • Place of Presentation: (Yokohama)
  • Year and Date: 2011-07-04
• [Presentation] 染色体分離の同期性を保証するメカニズム (2011)
  • Author(s): 広田 亨
  • Organizer: 北海道大学大学院・先端生命科学研究院・分子生物学特別セミナー
  • Place of Presentation: (札幌)
  • Year and Date: 2011-06-24
• [Presentation] How chromosome assembly is triggered in cells entering mitosis (2011)
  • Author(s): Hirota, T
  • Organizer: Impromptu Seminar at the IMP.
  • Place of Presentation: (Vienna)
  • Year and Date: 2011-04-14
• [Presentation] Does the spindle-assembly checkpoint monitor the presence of tension? (2011)
  • Author(s): Uchida, K.S.K., Hirota, T
  • Organizer: 第30回日本分子生物学会.ワークショップ
  • Place of Presentation: (横浜)
• [Presentation] Probing separase activity reveals its switch-like activation at the metaphase-to- anaphase transition (2011)
  • Author(s): Hirota, T
  • Organizer: 第30回日本分子生物学会.シンポジウム
  • Place of Presentation: (横浜)
• [Presentation] Dual role of separase during the metaphase-to-anaphase transition ensures chromosome segregation in mammalian cells (2011)
  • Author(s): Shindo, N., Hirota, T
  • Organizer: Annual meeting of American Society of Cell Biology
  • Place of Presentation: (Denver)
• [Presentation] Probing separase activity reveals its switch-like activation at the metaphase-to- anaphase transition (2011)
  • Author(s): Hirota, T
  • Organizer: The MEXT Priority Research International Symposium "Cell Division"
  • Place of Presentation: (Hakone)
• [Presentation] Probing separase activity reveals its switch-like activation at the metaphase-to- anaphase transition (2011)
  • Author(s): Hirota, T
  • Organizer: The 2nd Dynamic Kinetochore Workshop
  • Place of Presentation: (Vienna)
• [Presentation] がん細胞における異数性の発生要因を考える.要望講演 (2011)
  • Author(s): 広田 亨
  • Organizer: 第52回日本臨床細胞学会総会
  • Place of Presentation: (福岡)
• [Presentation] How chromosome assembly is triggered in cells entering mitosis (2011)
  • Author(s): Hirota, T
  • Organizer: 6th UK-Japan Cell Cycle Meeting
  • Place of Presentation: (Windermere)
• [Presentation] がん細胞における異数性の発生要因を考える (2011)
  • Author(s): 広田 亨
  • Organizer: 第52回日本臨床細胞学術総会（春期大会）(招待講演)
  • Place of Presentation: 福岡
• [Presentation] Probing separase activity reveals its switch-like activation at the metaphase-to-anaphase transition (2011)
  • Author(s): Toru Hirota
  • Organizer: MEXT International Symposium: Cell Division(招待講演)
  • Place of Presentation: Hakone
• [Presentation] The switch-like activation of separase ensures chromosome segregation (2011)
  • Author(s): Toru Hirota
  • Organizer: 6th GCOE International Symposium: Mitosis(招待講演)
  • Place of Presentation: Yokohama
• [Presentation] Probing separase activity reveals its switch-like activation at the metaphase-to-anaphase transition (2011)
  • Author(s): Toru Hirota
  • Organizer: 第34回日本分子生物学会年会(招待講演)
  • Place of Presentation: Yokohama
• [Presentation] Does the spindle-assembly checkpoint monitor the presence of tension? (2011)
  • Author(s): Kazuhiko Uchida, Toru Hirota
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: Yokohama
• [Presentation] Heterochromatin protein 1 is an essential component of the chromosomal passenger complex to prevent missegregation of chromosomes
  • Author(s): Hirota, T
• [Presentation] How cells ensure separation of sister chromatids in mitosis.
  • Author(s): Toru Hirota
  • Organizer: International Symposium on Molecular Medicine.
  • Place of Presentation: Suwon, South Korea
  • Invited
• [Presentation] Ｍ期プロテアーゼ・セパレースの活性プローブによって分かったこと
  • Author(s): 広田 亨
  • Organizer: 東京理科大学先端生命研水曜セミナー
  • Place of Presentation: 野田市
  • Invited
• [Presentation] がん細胞における異数性の発生要因を考える
  • Author(s): 広田 亨
  • Organizer: 第８回新潟横断的消化器疾患研究グループ研究会
  • Place of Presentation: 新潟市
  • Invited
• [Book] 総説「細胞分裂の仕組みに迫る―染色体分離のキー酵素separaseの活性の可視化と作用機序の解明」バイオサイエンスとインダストリー (2013)
  • Author(s): 進藤軌久、広田 亨
• [Book] 総説「Greatwallによるリン酸化バランス調節に基づくM期開始制御」 実験医学増刊号・ヒトと医学のステージへ拡大する細胞周期 (2013)
  • Author(s): 阿部優介、広田 亨
  • Publisher: 羊土社
• [Book] ヒトと医学のステージへ拡大する細胞周期．総説「Greatwallによるリン酸化バランス調節に基づくＭ期開始制御」 (2013)
  • Author(s): 阿部優介
  • Total Pages: 221
  • Publisher: 羊土社
• [Book] Review. Handbook of Proteolytic Enzymes. Third Edition, Edited by Rawlings and Salvesen, Elsevier. (2012)
  • Author(s): Kumada, K., and *Hirota, T
  • Publisher: Separase
• [Book] 記事「がん医療を拓く―ふぞろいを狙い撃つ」 (2012)
  • Author(s): 広田 亨
  • Publisher: ロハスメディカル
• [Book] 総説「バイオセンサーにより明らかにされた染色体分離におけるセパラーゼの異なる2つの役割」 (2012)
  • Author(s): 進藤軌久、広田 亨
  • Publisher: ライフサイエンス新着論文レビュー
• [Book] Encyclopedia of Life Sciences, John Wiley & Sons, Ltd (2011)
  • Author(s): Nagasaka, K., Gallego-Paez, L.M., * Hirota, T
  • Publisher: Mitosis and Meiosis: Molecular Control of Chromosome Separation. Review
• [Remarks] ホームページ
  • URL: http://www.jfcr.or.jp/tci/exppathol/index.html
• [Remarks] 公益財団法人がん研究会 実験病理部 広田亨研究室
  • URL: http://www.jfcr.or.jp/tci/exppathol/index.html
• [Remarks]
  • URL: http://www.jfcr.or.jp/tci/exppathol/index.html
• [Patent(Industrial Property Rights)] セパレース・バイオセンサー (2012)
  • Inventor(s): 広田亨、進藤軌久、熊田和貴
  • Industrial Property Rights Holder: 公益財団法人がん研究会
  • Filing Date: 2012
  • Overseas
• [Patent(Industrial Property Rights)] セハレース・バイオセンサーに関する国際特許 (2012)
  • Inventor(s): 広田亨、進藤軌久、熊田和貴
  • Industrial Property Rights Holder: 広田亨、進藤軌久、熊田和貴
  • Industrial Property Rights Type: 特許
  • Filing Date: 2012-07-24
  • Overseas