Single-molecule visualisation of kinetochore-microtubule interface using the in vitro reconstitution system
| Project/Area Number |
16K14672
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Molecular biology
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| Research Institution | Hiroshima University |
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Principal Investigator |
Toda Takashi 広島大学, 先端物質科学研究科, 特任教授 (50197894)
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| Co-Investigator(Kenkyū-buntansha) |
湯川 格史 広島大学, 先端物質科学研究科, 特任助教 (50403605)
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| Research Collaborator |
Matsuo Yuzy
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 染色体 / 紡錘体微小管 / in vitro再構成 / 微小管ポリメラーゼ / 微小管結合タンパク質 / 全反射照明蛍光顕微鏡 / 動原体 / 分裂酵母 / 紡錘体（スピンドル）微小管 / 動原体構成タンパク質 / 微小管重合 / 分裂装置 / キネトコア / 微小管 / in vitro / TIRF / スピンドル / 染色体分配 / 再構築 / TIRF顕微鏡 / １分子視覚化 |
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| Outline of Final Research Achievements |
Equal partition of genetic material (chromosomes) lies at the heart of cell reproduction. The kinetochore is a large proteinaceous structure formed on the chromosome. The microtubule is a major component of division apparatus. For each pair of chromosomes to segregate equally, the microtubule captures and pulls the kinetochore towards each pole. The aim of this program was the in vitro reconstitution of the interface that recapitulates the kinetochore-microtubule interaction. Using fission yeast, we first showed that Ndc80 (a core kinetochore component) and Dis1 (a microtubule polymerase) genetically interact. The following in vitro study indicated that these two proteins do not interact directly; instead, the third component, the third protein Mal3/EB1 is necessary for Ndc80 to interact with Dis1. These result have suggested that three factors, namely Ndc80, Dis1 and Mal3, comprise the minimal requirement for the in vitro reconstruction of the kinetochore-microtubule interface.
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| Academic Significance and Societal Importance of the Research Achievements |
染色体の娘細胞への均等分配は、生命の継承、生物の生存にとって最も重要なプロセスである。染色体不分離、非均等分配が起こると、生殖細胞では流産あるいはダウン症候群、常細胞では、細胞ガン化が惹起される。本研究により、染色体分配には３つのタンパク質が必要最小限であることが、明らかになった。この成果により、基礎生命科学分野におけて長年の謎であった染色体分配機構の全容解明への道程が示された。一方、医学、医療分野に対しては、３つのタンパク質をターゲットとする活性小分子の開発、創薬を可能とし、ヒト疾患の予防、治癒に寄与する。よって本研究成果は、学術的、社会的意義、両面から重要な知見を提供したと云える。
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Report
(4 results)
Research Products
(45 results)
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[Journal Article] A microtubule polymerase cooperates with the kinesin-6 motor and a microtubule cross-linker to promote bipolar spindle assembly in the absence of kinesin-5 and kinesin-14 in fission yeast2017
Author(s)
Yukawa, M., Kawakami, T., Okazaki, M., Kume, K., Tang, N.H., and Toda, T.
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Journal Title
Mol Biol Cell
Volume: 28
Pages: 3647-3659
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Journal Article] Generation and characterisation of temperature sensitive mutants of genes encoding the fission yeast spindle pole body2017
Author(s)
Tang, N.H., Fong, C.S., Jourdain, I., Masuda, H., Yukawa, M., and Toda, T.
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Journal Title
PeerJ Preprints
Volume: 5
Pages: e3377v1
DOI
Related Report
Open Access / Int'l Joint Research
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[Journal Article] Identification of three signaling molecules required for calcineurin-dependent monopolar growth induced by the DNA replication checkpoint in fission yeast2017
Author(s)
Kume, K., Hashimoto, T., Suzuki, M., Mizunuma, M., Toda, T., & Hirata, D.
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Journal Title
Biochem Biophys Res Commun
Volume: 491
Pages: 883-889
DOI
Related Report
Peer Reviewed
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[Journal Article] An unconventional interaction between Dis1/TOG and Mal3/EB1 in fission yeast promotes the fidelity of chromosome segregation2016
Author(s)
Matsuo, Y., Maurer, S. P., Yukawa, M., Zakian, S., Singleton, M. R., Surrey, T. and Toda,
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Journal Title
J Cell Sci
Volume: 129
Pages: 4592-4606
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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