Structural basis for small RNA functional mechanism in eukaryotes

• Project/Area Number: 25840019
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Niigata University
• Principal Investigator: Miyoshi Tomohiro 新潟大学, 研究推進機構超域学術院, 助教 (60534550)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: Argonaute / small RNA / 小分子RNA / 遺伝子発現制御 / X線結晶構造解析 / RNAサイレンシング / 小分子核酸 / non-coding RNA

Outline of Final Research Achievements

Argonaute proteins are widespread in all domains of life from bacteria to eukaryotes, and are an essential component of the nucleic acid-based silencing systems. Although many aspects of the structure of Argonaute-nucleic acid complexes have been described in previous crystal structure studies, the recognition mechanism of the guide and target strands by Argonaute proteins is still not fully understood. In the present study, we have solved the structure of Argonaute in complex with guide RNA and target DNA by X-ray crystallography.

Research Products

• [Journal Article] Crystal structure of translation initiation factor 5B from the crenarchaeon Aeropyrum pernix. (2016)
  • Author(s): Murakami R, Miyoshi T, Uchiumi T, Ito K.
  • Journal Title: Proteins Volume: in press Issue: 5 Pages: 712-717
  • DOI: 10.1002/prot.25009
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Functional role of the C-terminal tail of the archaeal ribosomal stalk in recruitment of two elongation factors to the sarcin/ricin loop of 23S rRNA. (2015)
  • Author(s): Imai, H, Miyoshi, T, Murakami, R, Ito, K, Ishino, Y, and Uchiumi, T.
  • Journal Title: Genes. Cells Volume: 20 Issue: 7 Pages: 613-624
  • DOI: 10.1111/gtc.12256
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Solution structure of human P1-P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome (2013)
  • Author(s): Lee, K.M., Yusa, K., Chu, L.O., Yu, C.W., Oono, M., Miyoshi, T., Ito, K., Shaw, P.C., Wong, K.B., and Uchiumi, T
  • Journal Title: Nucleic Acids Res Volume: 41 Issue: 18 Pages: 8776-8787
  • DOI: 10.1093/nar/gkt636
  • Peer Reviewed
• [Presentation] アルゴノート蛋白質によるガイド/ターゲット二本鎖の新結合様式 (2015)
  • Author(s): 三好智博、伊東孝介、村上僚、内海利男
  • Organizer: 第38回日本分子生物学学会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-01
• [Presentation] Recognition mechanism of guide RNA-target DNA heterodupulex by Argonaute (2015)
  • Author(s): Tomohiro Miyoshi, Kosuke Ito, Ryo Murakami, Toshio Uchiumi
  • Organizer: EMBL Symposium
  • Place of Presentation: EMBL Advanced Training Centre (Germany)
  • Year and Date: 2015-10-18
  • Int'l Joint Research
• [Presentation] Rhodobacter sphaeroides ArgonauteによるガイドRNA/ターゲットDNA認識機構の構造基盤 (2015)
  • Author(s): 三好智博、伊東孝祐、村上僚、内海利男
  • Organizer: 第17回日本RNA学会年会
  • Place of Presentation: ホテルライフォート札幌
  • Year and Date: 2015-07-15
• [Presentation] RNA-guided DNA interferenceを担うArgonauteタンパク質 (2015)
  • Author(s): 三好智博、伊東孝祐、村上僚、内海利男
  • Organizer: 平成27年度日本生化学会関東支部例会
  • Place of Presentation: 新潟日報メディアシップ
  • Year and Date: 2015-06-20
• [Presentation] リボトキシン作用に関与するリボソームP1/P2ダイマーの柔軟な機能構造 (2013)
  • Author(s): 遊佐和之、三好智博、Ka-Ming Lee、Lai-On Chu, Conny Wing-Heng Yu, 大野萌、伊東孝祐、Pang-Chui Shaw, Kam-Bo Wong、内海利男
  • Organizer: 第8回 無細胞生命科学研究会
  • Place of Presentation: 新潟大学