In-vitro reconstitution of mitotic chromosomes
| Project/Area Number |
18H02381
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Review Section |
Basic Section 43010:Molecular biology-related
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Shintomi Keishi 国立研究開発法人理化学研究所, 開拓研究本部, 専任研究員 (60462694)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
Fiscal Year 2020: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2019: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
Fiscal Year 2018: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
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| Keywords | 染色体 / 再構成 / トポⅡ / コンデンシン / カエル卵抽出液 / トポイソメラーゼ / トポII / トポイソメラーゼⅡ / 分裂期 / トポイソメラーゼII / ヌクレオソーム / DNAトポロジー |
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| Outline of Final Research Achievements |
Topoisomerase IIα (topo IIα) is one of the six proteins essential for mitotic chromatid reconstitution in vitro. It is not fully understood how this enzyme regulates chromosome assembly remains elusive. In this project, we found that chromosomal binding of topo IIα is sensitive to buffer conditions and depends on its C-terminal domain (CTD). Enzymological assays using circular DNA substrates supports the idea that topo IIα first resolves inter-chromatid entanglements to drive individualization and then generates intra-chromatid entanglements to promote thickening. Importantly, only the latter process requires the CTD. By using frog egg extracts, we also show that the CTD contributes to proper formation of nucleosome-depleted chromatids by counteracting anomalous aggregated . Our results demonstrate that topo IIα utilizes its CTD to deliver the enzymatic core to crowded environments created during mitotic chromatid assembly, thereby fine-tuning this process.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究成果は、細胞の分裂期におけるトポⅡαの役割を初めて明確に示したものであり、染色体の構造と機能を大きく進展させるものである。さらに、トポⅡαは抗がん剤の標的タンパク質として知られているだけでなく、核移植によってクローン個体を作出する技術との関連も示唆されていることから、実学的研究にも大きなインパクトを与えると期待される。
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Report
(4 results)
Research Products
(15 results)
