Structural analysis of modified NCP using mass spectrometry
| Project/Area Number |
17K07313
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Yokohama City University |
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Principal Investigator |
AKASHI Satoko 横浜市立大学, 生命医科学研究科, 教授 (10280728)
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| Project Period (FY) |
2017-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 質量分析 / ヌクレオソームコア粒子 / 構造解析 / 蛋白質 / 生体分子 / 分析科学 |
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| Outline of Final Research Achievements |
To clarify structural changes in nucleosomes caused by modifications, such as DNA methylation and histone acetylation, we performed biophysical analysis of modified nucleosome core particles (NCPs) that were reconstituted in vitro. In the analysis of modified NCPs, mono- and di-NCP were prepared with 147 and 366 base-pair (bp) DNA oligomers. Further, we prepared NCPs with DNA oligomers having AT or CG rich sequences, and characterized their biophysical properties by mass spectrometry and small angle X-ray scattering. No structural information is available for these NCPs due to difficulty in crystallization. We established a mass spectrometric method to reliably determine the methylation levels of DNA fragments with ~400 bp.
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| Academic Significance and Societal Importance of the Research Achievements |
極めて多くのサイトがメチル化されたDNAを用いて再構成されたDNAの構造解析のための要素技術を質量分析を駆使して開発できたことは、エピゲノム科学研究の基盤構築に貢献できると考えられる。特に、今後クライオ電顕による構造解析を組み合わせることで、このような研究が大幅に進展すると考えられるが、そのための準備段階として、有用な知見が得られた。
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Report
(5 results)
Research Products
(48 results)
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[Journal Article] Crystal structure of the overlapping dinucleosome composed of hexasome and octasome2017
Author(s)
Daiki Kato, Akihisa Osakabe, Yasuhiro Arimura, Yuka Mizukami, Naoki Horikoshi, Kazumi Saikusa, Satoko Akashi, Yoshifumi Nishimura, Sam-Yong Park, Jumpei Nogami, Kazumitsu Maehara, Yasuyuki Ohkawa, Atsushi Matsumoto, Hidetoshi Kono, Rintaro Inoue, Masaaki Sugiyama, Hitoshi Kurumizaka
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Journal Title
Science
Volume: 356
Issue: 6334
Pages: 205-208
DOI
Related Report
Peer Reviewed / Open Access / Acknowledgement Compliant
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