Identification and characterization of the RISC-loading complex

• Project/Area Number: 22687014
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Molecular biology
• Research Institution: The University of Tokyo
• Principal Investigator: TOMARI Yukihide 東京大学, 分子細胞生物学研究所, 教授 (90447368)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥24,700,000 (Direct Cost: ¥19,000,000、Indirect Cost: ¥5,700,000); Fiscal Year 2012: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000); Fiscal Year 2011: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000); Fiscal Year 2010: ¥9,490,000 (Direct Cost: ¥7,300,000、Indirect Cost: ¥2,190,000)
• Keywords: 核酸 / 蛋白質 / 発現制御 / 生体分子 / 遺伝子

Research Abstract

Small RNAs including siRNAs and microRNAs act through formation of RNA-induced silencing complex (RISC), the core component of which is Argonaute family proteins. Here we show that the Hsc70/Hsp90 chaperone machinery is essential for loading of small RNA duplexes into Argonaute. Moreover, we established an in vitro reconstitution system for RISC assembly composed of seven protein factors includingthe chaperone machinery.

Research Products

• [Journal Article] miRNA-like duplexes as RNAi triggers with improved specificity. (2012)
  • Author(s): Betancur JG, Yoda M.
  • Journal Title: Front Genet. Volume: 3(127) Pages: 127-127
  • DOI: 10.3389/fgene.2012.00127
  • Peer Reviewed
• [Journal Article] microRNAs mediate gene silencing via multiple different pathways in Drosophila. (2012)
  • Author(s): Fukaya T, *Tomari Y.
  • Journal Title: Mol Cell. Volume: 48(6) Issue: 6 Pages: 825-836
  • DOI: 10.1016/j.molcel.2012.09.024
  • Peer Reviewed
• [Journal Article] Dicer is dispensable for asymmetric RISC loading in mammals (2012)
  • Author(s): Betancur JG, et al.
  • Journal Title: RNA Volume: 18 Issue: 1 Pages: 24-30
  • DOI: 10.1261/rna.029785.111
  • Peer Reviewed
• [Journal Article] Native gel analysis for RISC assembly (2011)
  • Author(s): Kawamata T, *To mari Y
  • Journal Title: Methods Mol Biol Volume: 725 Pages: 91-105
  • Peer Reviewed
• [Journal Article] Recognition of the pre-miRNA structure by Drosophila Dicer-1 (2011)
  • Author(s): Tsutsumi A, Kawamata T, Izumi N, Seitz H, *Tomari Y
  • Journal Title: Nat Struct Mol Biol Volume: 18 Pages: 1153-1158
  • Peer Reviewed
• [Journal Article] Multilayer checkpoints for microRNA authenticity during RISC assembly (2011)
  • Author(s): 川俣朋子、依田真由子、泊幸秀
  • Journal Title: EMBO Reports Volume: 12(9) Issue: 9 Pages: 944-949
  • DOI: 10.1038/embor.2011.128
  • Peer Reviewed
• [Journal Article] Native Gel Analysis for RISC Assembly (2011)
  • Author(s): 川俣朋子、泊幸秀
  • Journal Title: Methods in Molecular Biology Volume: 725 Pages: 91-105
  • DOI: 10.1007/978-1-61779-046-1_7
  • ISBN: 9781617790454, 9781617790461
  • Peer Reviewed
• [Journal Article] Making RISC. (2010)
  • Author(s): Kawamata T, *Tomari Y
  • Journal Title: Trends Biochem Sci Volume: 35 Pages: 368-376
  • Peer Reviewed
• [Journal Article] Hsc70/Hsp90 chaperone machinery mediates ATP-dependent RISC loading of small RNA duplexes (2010)
  • Author(s): Iwasaki S, Kobayashi M, Yoda M, Sakaguchi Y, Katsuma S, Suzuki T, Tomari Y
  • Journal Title: Mol Cell Volume: 39 Pages: 292-299
  • Peer Reviewed
• [Journal Article] Hsc70/Hsp90 chaperone machinery mediates ATP-dependent RISC loading of small RNA duplexes. (2010)
  • Author(s): Iwasaki S, et al.
  • Journal Title: Mol Cell. Volume: 39 Pages: 292-299
  • Peer Reviewed
• [Presentation] How to make RISC (2012)
  • Author(s): 泊幸秀
  • Organizer: FURANO Conference
  • Place of Presentation: 富良野市(招待講演)
  • Year and Date: 2012-03-06
• [Presentation] RNAサイレンシング複合体の形成と機能 (2012)
  • Author(s): 泊 幸秀
  • Organizer: アンチセンス・遺伝子・デリバリー合同シンポジウム
  • Place of Presentation: 仙台市民会館、仙台市
  • Invited
• [Presentation] In vitro reconstitution of human RISC assembly/ヒトRISC形成過程の再構成 (2012)
  • Author(s): 成瀬 健
  • Organizer: 第35回分子生物学会年会
  • Place of Presentation: 福岡国際会議場、福岡市
• [Presentation] Chaperoning RISC assembly (2011)
  • Author(s): 泊幸秀
  • Organizer: 長崎大学NEKKEN Special Seminar
  • Place of Presentation: 長崎市(招待講演)
  • Year and Date: 2011-11-21
• [Presentation] Chaperoning RISC Assembly (2011)
  • Author(s): Tomari Y.
  • Organizer: Keystone Symosia Mechanism and Biology of Silencing
  • Place of Presentation: Monterey (USA)
  • Year and Date: 2011-03-22
• [Presentation] Chaperoning RISC Assembly. (2011)
  • Author(s): Tomari Y
  • Organizer: Keystone Symposia Mechanism and Biology of Silencing
  • Place of Presentation: Monterey,USA
• [Presentation] In vitro Reconstitution of RISC Assembly Mediated by Hsc70/Hsp 90 Chaperone Machinery (2011)
  • Author(s): Iwasaki S, Tomari Y
  • Organizer: The 16th Annual Meeting of RNA Society
  • Place of Presentation: Kyoto
• [Presentation] Chaperoning RISC Assembly (2010)
  • Author(s): Tomari Y.
  • Organizer: The 2^<nd> Korean Oligonucleotide Therapeutics Society International Symposium
  • Place of Presentation: Seoul (Korea)
  • Year and Date: 2010-11-30
• [Presentation] Hsc70/Hsp90シャペロンマシナリーによるRISC loading (2010)
  • Author(s): 岩崎信太郎
  • Organizer: 第12回日本RNA学会
  • Place of Presentation: 一ツ橋記念講堂(東京)
  • Year and Date: 2010-07-27
• [Presentation] Chaperone Machinery is Required for RISC-loading (2010)
  • Author(s): Iwasaki S, Tomari Y
  • Organizer: 19th CDB Meeting /RNA Sciences in Cell and Developmental Biology
  • Place of Presentation: Riken CDB, Kobe, Japan
• [Presentation] Chaperone Machinery is Required for RISC-loading (2010)
  • Author(s): Iwasaki S.
  • Organizer: 19^<th> CDB Meeting/RNA Sciences in Cell and Developmental Biology
  • Place of Presentation: 理化学研究所・発生科学総合研究センター(神戸)