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Reconstruction of in vitro piRNA biogenesis using silkworm germ line BmN4 cells

Research Project

Project/Area Number 25840041
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Functional biochemistry
Research InstitutionThe University of Tokyo

Principal Investigator

NISHIDA Kazumichi  東京大学, 理学(系)研究科(研究院), 助教 (10598436)

Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
KeywordspiRNA生合成 / PIWIタンパク質 / RNAヘリカーゼタンパク質 / 生殖細胞 / カイコ / piRNA / トランスポゾン / BmVasa
Outline of Final Research Achievements

piRNAs associate with PIWI proteins and silence selfish transposable elements. piRNA biogenesis consists of two sequential steps; primary piRNA processing and the Ping-Pong cycle. However, piRNA biogenesis remains elusive. We aimed to elucidate about piRNA pathway.
A Bombyx mori germ cell line, BmN4, is an excellent tool to investigate the molecular pathways of piRNA biogenesis. BmN4 expresses two PIWI proteins (Siwi and BmAgo3). We show that Siwi binds with BmVasa. BmVasa is DEAD-box RNA helicase protein. A lack of BmVasa abolishes the Ping-Pong cycle. Also, Siwi is associated with BmSpn-E. BmSpn-E, unlike BmVasa, is necessary for primary piRNA production. We propose a new model for piRNA biogenesis, where the BmSpn-E function in primary piRNA processing, whereas BmVasa is important for the Ping-Pong cycle.

Report

(3 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • Research Products

    (5 results)

All 2015 2014 2013 Other

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results,  Acknowledgement Compliant: 1 results) Presentation (4 results)

  • [Journal Article] Respective functions of two distinct Siwi complexes assembled during piRNA biogenesis in insect germ cells.2015

    • Author(s)
      Nishida, KM., Iwasaki, YW., Murota, Y., Nagao, A., Mannen, T., Kato, Y., Siomi, H., and Siomi, MC.
    • Journal Title

      Cell Reports

      Volume: 10 Issue: 2 Pages: 193-203

    • DOI

      10.1016/j.celrep.2014.12.013

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] Respective functions of two distinct Siwi, a silkworm PIWI, complexes assembled during piRNA biogenesis in insect germ cells2014

    • Author(s)
      Kazumichi M. Nishida, Yuka W. Iwasaki, Yukiko Murota, Taro Mannen, Haruhiko Siomi and Mikiko C. Siomi
    • Organizer
      日本分子生物学会
    • Place of Presentation
      パシフィコ横浜 (横浜)
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Annual Research Report
  • [Presentation] Respective functions of two distinct Siwi, a silkworm PIWI, complexes assembled during piRNA biogenesis in insect germ cells2014

    • Author(s)
      Kazumichi M. Nishida, Yuka W. Iwasaki, Yukiko Murota, Taro Mannen, Haruhiko Siomi and Mikiko C. Siomi
    • Organizer
      joint Australia and Japan RNA
    • Place of Presentation
      University o Technology Sydney (Australia)
    • Year and Date
      2014-11-02 – 2014-11-05
    • Related Report
      2014 Annual Research Report
  • [Presentation] カイコpiRNA生合成経路におけるBmQinの機能解析2013

    • Author(s)
      西田 知訓
    • Organizer
      第15回日本RNA学会年会
    • Place of Presentation
      ひめぎんホール(愛媛県)
    • Related Report
      2013 Research-status Report
  • [Presentation] BmVasa involve discharge of cleavage product from piRISC in silkworm piRNA biogenesis

    • Author(s)
      西田 知訓
    • Organizer
      第14回 東京大学生命科学シンポジウム
    • Place of Presentation
      東京都
    • Related Report
      2013 Research-status Report

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Published: 2014-07-25   Modified: 2019-07-29  

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