Analysis of piRNA biogenesis pathway

• Project/Area Number: 17H03632
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Keio University (2018-2019); The University of Tokyo (2017)
• Principal Investigator: ISHIZU Hirotsugu 慶應義塾大学, 医学部(信濃町), 講師 (40574588)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2019: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2018: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2017: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
• Keywords: piRNA / Armi / Piwi / Zuc / GasZ / レトロトランスポゾン / RNAサイレンシング / CLIPPARE / Yb / RNA silencing / Armitage / iCLIP / RNA helicase

Outline of Final Research Achievements

piRNAs are germ cell-specific functional small molecule RNAs that repress the expression of genes with complementary sequences. We identified an ATP-dependent RNA helicase, Armitage (Armi), as a piRNA biogenesis factor in Drosophila. By analyzing the RNA that binds to Armi in cells using iCLIP and CLIPPARE that we independently developed, we were able to capture the transition process of piRNA biogenesis, which has not been analyzed in detail. Biochemical analysis revealed a detailed molecular mechanism of the piRNA biogenesis pathway.

Academic Significance and Societal Importance of the Research Achievements

piRNA生合成経路は、関連因子が多い複雑な分子機構であり、その中間過程に関する知見が全容解明には不可欠である。本研究では、近年開発された技術を取り入れ、関連因子の細胞内局在と機能、プロセシング過程の中間体配列に関する解析を行い、piRNA生合成経路の包括的な理解を深め、成果を上げてきた。さらに、本研究ではCLIP法を応用した新たな配列解析手法を確立した。この新たな手法による中間体解析が、今後もpiRNA生合成経路の全容解明に寄与すると期待している。

Research Products

• [Journal Article] Distinct and collaborative functions of Yb and Armitage in transposon-targeting piRNA biogenesis (2019)
  • Author(s): Hirotsugu Ishizu, Tatsuki Kinoshita, Shigeki Hirakata, Chihiro Komatsuzaki, and Mikiko C. Siomi
  • Journal Title: Cell Reports Volume: 27 Issue: 6 Pages: 1822-1835
  • DOI: 10.1016/j.celrep.2019.04.029
  • Peer Reviewed / Open Access
• [Journal Article] Requirements for multivalent Yb body assembly in transposon silencing in Drosophila (2019)
  • Author(s): Shigeki Hirakata, Hirotsugu Ishizu, Aoi Fujita, Yumiko Tomoe, Mikiko C. Siomi
  • Journal Title: EMBO Reports Volume: 印刷中 Issue: 7
  • DOI: 10.15252/embr.201947708
  • Peer Reviewed / Open Access
• [Journal Article] Nuclear RNA export factor variant initiates piRNA-guided co-transcriptional silencing (2019)
  • Author(s): Murano, K., Iwasaki Y.W., Ishizu H., Mashiko, A., Shibuya, A., Kondo, S., Adachi, S., Suzuki, S., Saito, K., Natsume, T., Siomi, M.C., Siomi, H.
  • Journal Title: The EMBO Journal Volume: 印刷中 Issue: 17 Pages: 1-20
  • DOI: 10.15252/embj.2019102870
  • Peer Reviewed / Open Access
• [Journal Article] Armitage determines PiwiーpiRISC processing from precursor formation and quality control to inter‐organelle translocation (2019)
  • Author(s): Yamashiro Haruna、Negishi Mayu、Kinoshita Tatsuki、Ishizu Hirotsugu、Ohtani Hitoshi、Siomi Mikiko C
  • Journal Title: EMBO reports Volume: 21 Issue: 2
  • DOI: 10.15252/embr.201948769
  • Peer Reviewed / Open Access
• [Journal Article] Distinct and collaborative functions of Yb and Armitage in transposon-targeting piRNA biogenesis (2019)
  • Author(s): Ishizu, H., Kinoshita, T., Hirakata, S., Komatsuzaki, C., Siomi, M. C.
  • Journal Title: Cell Reports Volume: 印刷中
  • Peer Reviewed / Open Access
• [Journal Article] The nucleolar transcriptome regulates Piwi shuttling between the nucleolus and the nucleoplasm (2018)
  • Author(s): Mikhaleva, E. A., Leinsoo, T.A., Ishizu, H., Gvozdev, V. A., Klenov, M.S.
  • Journal Title: Chromosome Research Volume: 27 Issue: 1-2 Pages: 141-152
  • DOI: 10.1007/s10577-018-9595-y
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Use of the CRISPR-Cas9 system for genome editing in cultured Drosophila ovarian somatic cells. (2017)
  • Author(s): Ishizu H, Sumiyoshi T, Siomi MC
  • Journal Title: Methods Volume: 126 Pages: 186-192
  • DOI: 10.1016/j.ymeth.2017.05.021
  • Peer Reviewed / Open Access
• [Presentation] Role of the RNA helicase, Armitage, in the primary piRNA biogenesis pathway (2019)
  • Author(s): Hirotsugu Ishizu, Tatsuki Kinoshita, MIkiko C. Siomi
  • Organizer: Keystone symposia, Small Regulatory RNAs
  • Int'l Joint Research
• [Presentation] ショウジョウバエ卵巣体細胞における一次piRNA整合性経路の解析 (2019)
  • Author(s): 石津大嗣、塩見春彦、塩見美喜子
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] Role of the RNA hilicase, Armitage, in the primary piRNA biogenesis pathway (2019)
  • Author(s): Hirotsugu Ishizu
  • Organizer: Keystone Symposia, Small Regulatory RNAs
  • Int'l Joint Research
• [Presentation] 一次piRNA生合成経路におけるRNAヘリカーゼArmitageの機能 (2018)
  • Author(s): 石津大嗣
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] Role of the RNA helicase, Armitage, in the primary piRNA biogenesis pathway (2017)
  • Author(s): Ishizu, H., Kinoshita, T., Siomi, MC
  • Organizer: The 43rd Naito Conference on “NoncodingRNA: Biology, Chemistry, & Diseases”
  • Int'l Joint Research
• [Presentation] piRNA生合成経路によるmRNA分解制御 (2017)
  • Author(s): 石津大嗣、塩見美喜子
  • Organizer: 第19回日本RNA学会年会
• [Remarks] 慶應義塾大学医学部分子生物学教室のwebサイト
  • URL: http://siomilab.med.keio.ac.jp/
• [Remarks] 東京大学理学部生物化学科塩見研究室ホームページ
  • URL: http://www-siomilab.biochem.s.u-tokyo.ac.jp/index.html