Identification of the role of transcription termination and polyadenylation in the pathogenesis of ALS using multiple high-throughput sequencing analyses
| Project/Area Number |
15K06755
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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 |
Nerve anatomy/Neuropathology
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| Research Institution | Nagoya University |
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Principal Investigator |
Masuda Akio 名古屋大学, 医学系研究科, 准教授 (10343203)
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| Project Period (FY) |
2015-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | tRIP / CLIP法 / RNA代謝 / ポリアデニル化 / RNA-タンパク相互作用 / ALS / FUS / RNA-蛋白相互作用 / CLIP / RNAP II / mRNA / 次世代シークエンサー |
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| Outline of Final Research Achievements |
RNA metabolism occurs in specific machineries of RNA, including RNA polymerase II (RNAP II), spliceosome, and 3’ end processing machinery. However, the transcriptome-wide detection of protein-RNA interactions in a subcellular component is challenging. Here, we developed the tRIP method (targeted RNA immunoprecipitation) to increase efficiency of RNA immunoprecipitation, which enables detection of UV-crosslinked protein-RNA interactions from thousands of cells. Application of tRIP to identification of protein-protein-RNA interactions revealed that FUS, a multifunctional RNA-binding protein associated with ALS, binds specifically upstream to alternative polyadenylation (APA) sites of nascent RNA engaged to RNAP II. Further tRIP analysis demonstrated that the FUS-U1 snRNP-RNA complex is the entity that suppresses APA.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究は、生命の基本現象である、遺伝・転写・翻訳のうち、転写に関するものである。本研究により、従来の100倍以上の感度を持った転写制御解析法を確立した。この方法により、従来、解析することの出来なかった、細胞内の小器官内での転写の様子を見ることが可能になった。実際、本研究で、疾患原因因子FUSによる、従来知られていなかった新たな転写制御様式を発見している。本研究成果は、転写研究や転写異常が原因となる神経疾患の研究の進展に大いに貢献することが期待される。
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Report
(5 results)
Research Products
(38 results)
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[Journal Article] R-spondin 2 promotes acetylcholine receptor clustering at the neuromuscular junction via Lgr5.2016
Author(s)
Nakashima H., Ohkawara B., Ishigaki S., Fukudome T., Ito K., Tsushima M., Konishi H., Okuno T., Yoshimura T., Ito M., Masuda A., Sobue G., Kiyama H., Ishiguro N., Ohno K.
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Journal Title
Sci Rep
Volume: 22
Issue: 1
Pages: 28512-28512
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Arginine methylation of ubiquitin-associated protein 2-like is required for the accurate distribution of chromosomes.2016
Author(s)
Maeda M, Hasegawa H, Sugiyama M, Hyodo T, Ito S, Chen D, Asano E, Masuda A, Hasegawa Y, Hamaguchi M, Senga T
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Journal Title
FASEB Journal
Volume: 30
Issue: 1
Pages: 312-323
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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