| Project/Area Number |
17K19335
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Molecular and Genome biology and related fields
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| Research Institution | Hokkaido University |
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Principal Investigator |
Hirose Tetsuro 北海道大学, 遺伝子病制御研究所, 教授 (30273220)
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| Project Period (FY) |
2017-06-30 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
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| Keywords | noncoding RNA / 液相分離 / RNA結合タンパク質 / 液体相分離 / ノンコーディングRNA / 液体相転移 / プリオン様ドメイン / 蛋白質 / 核酸 |
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| Outline of Final Research Achievements |
This study aims to develop an in vitro LLPS system and understand the LLPS mechanism using the system. We found that LLPS was induced with the RNA fragment derived from the C2 subdomain of NEAT1 arcRNA, that was identified as the functional region for nuclear paraspeckle formation by CRISPR/Cas9 genome editing analysis, which was synthesized in vitro and mixed with HeLa nuclear extract. We devised sensitive and quantitative detection system of LLPS. Using the in vitro system, we identified NONO and SFPQ, which are required for paraspeckle assembly in vivo, are also required for in vitro LLPS and further identified RBM14 and FUS that are secondarily recruited to the C2 RNA. Thus, our research unveiled the mechanism of LLPS which was induced by C2 subdomain of NEAT1 arcRNA.
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| Academic Significance and Societal Importance of the Research Achievements |
LLPS研究は世界的に大きな注目を集め、多くの研究が急速に進んでいる。本研究は、そうした中でLLPSにRNAが果たす役割を解析するためのin vitro解析系を開発し、それを用いて実際に関与するタンパク質因子を同定したことが意義深い。さらにin vivo解析で同定したncRNAの機能ドメインの役割が、LLPS誘導機能であることを明確に示したこともncRNA研究分野に大きなインパクトを与える成果といえる。
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