Suppression of the splicing mutation based on the molecular mechanism of pre-mRNA splicing fidelity

• Project/Area Number: 25460057
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Hokkaido University
• Principal Investigator: MAITA Hiroshi 北海道大学, 薬学研究院, 講師 (60431318)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: スプライシング / ケミカルバイオロジー / 低分子化合物 / スプライソソーム / fidelity / スプライス部位変異 / snRNP / スプリットルシフェラーゼ

Outline of Final Research Achievements

We have developed an assay to find a compound that affect the spliceosome and have obtained several hit compounds by the automated screening of a small compound library. In this study we made a genetic reporter to detect the compound-mediated recovery of defected splicing that is caused by a mutation in the 3’ splice site. Some of our hit compounds have shown that they can induce splicing of the mutated intron at the correct site.

Research Products

• [Journal Article] A Splicing Reporter Tuned to Non-AG Acceptor Sites Reveals that Luteolin Enhances the Recognition of Non-canonical Acceptor Sites. (2016)
  • Author(s): Msanori Chiba, Hiroyoshi Ariga, Hiroshi Maita
  • Journal Title: Chemical Biology and Drug Design Volume: 87 Issue: 2 Pages: 275-282
  • DOI: 10.1111/cbdd.12656
  • NAID: 120006581491
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A split luciferase-based reporter for detection of a cellular macromolecular complex. (2014)
  • Author(s): Hiroshi Maita, Kenji Tomita, Hiroyoshi Ariga
  • Journal Title: Analytical Biochemistry Volume: 452 Pages: 1-9
  • DOI: 10.1016/j.ab.2014.01.015
  • NAID: 120005460254
  • Peer Reviewed
• [Presentation] Identification of small compounds that affect the cellular snRNP levels (2016)
  • Author(s): 米田 宏
  • Organizer: RNA2016
  • Place of Presentation: 京都国際会館（京都府京都市）
  • Year and Date: 2016-06-29
  • Int'l Joint Research
• [Presentation] アクセプター部位変異存在下でスプライシングを回復させる化合物の探索 (2015)
  • Author(s): 米田 宏
  • Organizer: 日本RNA学会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2015-07-15
• [Presentation] スプライシング異常を改善する生薬由来化合物の探索 (2015)
  • Author(s): 千葉 政徳、有賀 寛芳、米田 宏
  • Organizer: 日本薬学会第135年会
  • Place of Presentation: 兵庫県神戸市、神戸学院大学
  • Year and Date: 2015-03-27
• [Presentation] Screening of compounds affecting spliceosome formation by a split luciferase-based snRNP reporter (2013)
  • Author(s): 冨田 健司、有賀 寛芳、米田 宏
  • Organizer: 日本分子生物学会年会
  • Place of Presentation: 兵庫県神戸市、神戸国際展示場