Rapid protein degradation reveals functional coupling between the MCM helicase and cohesin, and their pleiotropic functions

• Project/Area Number: 17K15068
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Molecular biology
• Research Institution: National Institute of Genetics
• Principal Investigator: NATSUME Toyoaki 国立遺伝学研究所, 遺伝メカニズム研究系, 助教 (10435513)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2018: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2017: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: オーキシンデグロン法 / MCM2-7ヘリカーゼ / SMC複合体 / コヒーシン / コンデンシン / SMC5/6 / トポロジカルストレス / MCMヘリカーゼ / SMC5/6複合体 / 細胞周期特異的分解 / タンパク質分解 / 細胞周期

Outline of Final Research Achievements

It is of critical importance for organisms to precisely transmit chromosomal DNA to daughter cells and offsprings for the maintenance of life and species. In this study, by taking advantage of the auxin-inducible degron technology in human cells that enables rapid inactivation of target genes, I studied two important steps in chromosome transmission, DNA replication and chromosome segregation, with unprecedented time-resolution observation, and gave some critical and interesting findings. Especially, this study made a major breakthrough in the study of the SMC5/6 complex, whose role is still controversial in the world, giving foundations of future studies .

Academic Significance and Societal Importance of the Research Achievements

染色体DNAの伝承は、生命科学において重要な課題である一方、この過程の異常は、癌や様々な遺伝性疾患と密接な関わりがある。特に最近、幾つかの遺伝性疾患、癌、母年齢効果が、コヒーシンやSMC5/6の異常によって引き起こされることが分かってきた。本研究では、私が以前確立したヒト細胞におけるオーキシンデグロン法を用いて、コヒーシンに比べて研究が遅れているSMC5/6の基礎的な理解が大きく進んだ。今度は、患者のSMC5/6遺伝子の変異を含めた解析を進めることにより、これらの疾患の分子機構の解明及び治療に繋がると考えている。

Research Products

• [Journal Article] Chk1-mediated Cdc25A degradation as a critical mechanism for normal cell-cycle progression (2019)
  • Author(s): Goto H., Natsume T., Kanemaki M.T., Kaito A., Wang S., Gabazza E.C., Inagaki M., Mizoguchi A.
  • Journal Title: Journal of Cell Science Volume: 132
  • DOI: 10.1242/jcs.223123
  • Peer Reviewed / Open Access
• [Journal Article] Inhibiting the MCM8-9 complex selectively sensitizes cancer cells to cisplatin and olaparib (2019)
  • Author(s): Morii, I., Iwabuchi, Y., Mori, S., Suekuni, M., Natsume, T., Yoshida, K., Sugimoto, N., Kanemaki, MT. and Fujita, M.
  • Journal Title: Cancer Science Volume: 110 Pages: 1044-1053
  • DOI: 10.1111/cas.13941
  • Peer Reviewed / Open Access
• [Journal Article] Generation of conditional auxin-inducible degron (AID) cells and tight control of degron-fused proteins using the degradation inhibitor auxinole (2019)
  • Author(s): Yesbolatova A, Natsume T, Hayashi K, Kanemaki MT
  • Journal Title: Methods Volume: 印刷中 Pages: 73-80
  • DOI: 10.1016/j.ymeth.2019.04.010
  • Peer Reviewed / Open Access
• [Journal Article] Dynein-Dynactin-NuMA clusters generate cortical spindle-pulling forces as a multi-arm ensemble (2018)
  • Author(s): Okumura M, Natsume T, Kanemaki MT, Kiyomitsu T
  • Journal Title: eLife Volume: 7
  • DOI: 10.7554/elife.36559.001
  • Peer Reviewed / Open Access
• [Journal Article] Xrn2 accelerates termination by RNA polymerase II, which is underpinned by CPSF73 activity (2018)
  • Author(s): Eaton Joshua D.、Davidson Lee、Bauer David L.V.、Natsume Toyoaki、Kanemaki Masato T.、West Steven
  • Journal Title: Genes & Development Volume: 32 Pages: 127-139
  • DOI: 10.1101/gad.308528.117
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Ki-67 and condensins support the integrity of mitotic chromosomes through distinct mechanisms (2018)
  • Author(s): Takagi Masatoshi、Ono Takao、Natsume Toyoaki、Sakamoto Chiyomi、Nakao Mitsuyoshi、Saitoh Noriko、Kanemaki Masato T.、Hirano Tatsuya、Imamoto Naoko
  • Journal Title: J. Cell Sci. Volume: 131 Pages: 212092-212092
  • DOI: 10.1242/jcs.212092
  • Peer Reviewed / Open Access
• [Journal Article] Acute inactivation of the replicative helicase in human cells triggers MCM8-9-dependent DNA synthesis (2017)
  • Author(s): Natsume T, Nishimura K, Minocherhomji S, Bhwmick R, Hickson ID, Kanemaki MT
  • Journal Title: Genes Dev. Volume: advanced online Pages: 816-829
  • DOI: 10.1101/gad.297663.117
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Conditional Degrons for Controlling Protein Expression at the Protein Level (2017)
  • Author(s): Natsume Toyoaki、Kanemaki Masato T.
  • Journal Title: Annual Review of Genetics Volume: 51 Pages: 83-102
  • DOI: 10.1146/annurev-genet-120116-024656
  • Peer Reviewed
• [Presentation] 染色体制御におけるヒトSMC5/6の独自な役割を探る (2019)
  • Author(s): 夏目 豊彰
  • Organizer: 第36回染色体ワークショップ・第17回核ダイナミクス研究会
• [Presentation] ヒト染色体の維持における第三のSMC複合体「SMC5/6」の独自な機能を探る (2018)
  • Author(s): 夏目 豊彰
  • Organizer: 日本遺伝学会第90回大会
• [Presentation] Elucidating the unique and essential role of human SMC5/6, the yet-to-be-named SMC complex (2018)
  • Author(s): Toyoaki Natsume
  • Organizer: The 11th 3R&3C Symposium
  • Int'l Joint Research
• [Presentation] ゲノムの安定性を維持するためのヒトMCM8-9依存的な複製フォークの再生機構 (2018)
  • Author(s): 夏目 豊彰
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] Acute degradation of the human SMC complexes using the auxin-inducible degron (AID) technology reveals their distinct cell-cycle specific roles (2017)
  • Author(s): Natsume Toyoaki
  • Organizer: SMC Proteins 2017
  • Int'l Joint Research
• [Presentation] Natsume Toyoaki (2017)
  • Author(s): Acute degradation of the human SMC complexes using the auxin-inducible degron (AID) technology reveals their cell-cycle specific roles in maintaining genome stabilit
  • Organizer: Eukaryotic DNA Replication & Genome Maintenance
  • Int'l Joint Research