• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Biochemical approaches to understanding the reaction platforms of the piRNA pathway

Research Project

Project/Area Number 18H05271
Research Category

Grant-in-Aid for Scientific Research (S)

Allocation TypeSingle-year Grants
Review Section Broad Section G
Research InstitutionThe University of Tokyo

Principal Investigator

泊 幸秀  東京大学, 定量生命科学研究所, 教授 (90447368)

Co-Investigator(Kenkyū-buntansha) 中川 真一  北海道大学, 薬学研究院, 教授 (50324679)
Project Period (FY) 2018-06-11 – 2023-03-31
Project Status Granted (Fiscal Year 2022)
Budget Amount *help
¥193,570,000 (Direct Cost: ¥148,900,000、Indirect Cost: ¥44,670,000)
Fiscal Year 2022: ¥32,370,000 (Direct Cost: ¥24,900,000、Indirect Cost: ¥7,470,000)
Fiscal Year 2021: ¥32,370,000 (Direct Cost: ¥24,900,000、Indirect Cost: ¥7,470,000)
Fiscal Year 2020: ¥32,370,000 (Direct Cost: ¥24,900,000、Indirect Cost: ¥7,470,000)
Fiscal Year 2019: ¥51,350,000 (Direct Cost: ¥39,500,000、Indirect Cost: ¥11,850,000)
Fiscal Year 2018: ¥45,110,000 (Direct Cost: ¥34,700,000、Indirect Cost: ¥10,410,000)
KeywordspiRNA / RNAサイレンシング / PIWI / 小分子RNA / GTSF1 / 反応場 / 細胞内構造体 / nuage / P-body / ミトコンドリア / Zucchini / Trimmer
Outline of Annual Research Achievements

これまでに、遺伝学的スクリーニングと、タンパク質-タンパク質相互作用の探索によって、膨大な数のpiRNA関連因子が存在することが明らかになっている、その中で生化学活性が特定されているものは、我々が発見したpiRNA成熟のためのエクソヌクレアーゼTrimmerや、その前段階ではたらくエンドヌクレアーゼZucchiniを含めてごく少数に限られている。我々は、アメリカの研究室との共同研究と独自の生化学系の利用によって、GTSF1 (Gametocyte Specific Factor 1)と呼ばれる因子が、PIWIタンパク質と相互作用し、その標的RNA切断活性を大きく向上させる機能を持っているということを見いだした。さらに、別のアメリカの研究室との共同研究により、RNase κ と呼ばれるミトコンドリアに局在するヌクレアーゼが、piRNAの効率的な生産に重要であることを発見した。また、piRNA関連の新規因子を探索する解析を通じて、カイコが持つ2つのPIWIタンパク質、SiwiとBmAgo3のうち、BmAgo3をノックダウンした時だけ、Siwiと特異的に相互作用する機能未知の新規因子を発見した。さらには、次世代シーケンサーを駆使することにより、カイコBmN4細胞ゲノムの中に存在する最も代表的なpiRNAクラスターであるtorimochiの配列および性状解析を進めた。また、piRNA経路と同様に、トランスポゾンやウイルスなどの遺伝的侵略者からゲノムを守る役割を果たしているsiRNA経路については、その生合成において重要な働き果たすDicer(ショウジョウバエ由来のDicer-2)が長い二本鎖RNAを切断しながらsiRNAを次々と産生してゆく過程を一分子レベルで観察することに成功し、従来提案されていたモデルに修正を与えることとなった。

Current Status of Research Progress
Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

piRNA経路に関連するとされる膨大な数の因子の中で、生化学的な機能がきちんと同定されたものはごく一部に限られている。RNase κ は、piRNA生合成にかかわることが新規に見いだされたヌクレアーゼであり、piRNA生合成の生化学的理解大きく前進させるものである。さらに、GTSF1は、PIWIタンパク質も含めた大きなArgonauteファミリータンパク質のRNA切断活性を制御する初めての補助因子であり、その意義は大きい。また、piRNA経路と関連が深いsiRNA経路についても、一分子イメージングを駆使した大きな研究の進展が見られ論文として発表した。

Strategy for Future Research Activity

カイコに複数存在するGTSF1のホモログ(Gtsf1およびGtsf1-like)と、2つのPIWIタンパク質ホモログ(SiwiとAgo3)との間に存在する生化学的関係性を検証し、その協調的作用がpiRNA産生に与える影響を解析する。さらに、Siwiをノックダウンした際にのみ特異的にAgo3に結合する新規因子についても、そのpiRNApiRNA産生に与える影響や、相互作用する因子の探索を行う。また、カイコBmN4細胞の代表的なpiRNAクラスターであるtorimochiについては、複数種類の次世代シーケンサーを組み合わせることにより、周辺領域も含めた正確な配列の決定および性状解析を行い、他のトランスポゾンの配列や性状と大規模に比較する。

Assessment Rating
Interim Assessment Comments (Rating)

A: In light of the aim of introducing the research area into the research categories, the expected progress has been made in research.

Report

(8 results)
  • 2021 Annual Research Report
  • 2020 Abstract (Interim Assessment) ( PDF )   Annual Research Report   Interim Assessment (Comments) ( PDF )
  • 2019 Annual Research Report
  • 2018 Abstract ( PDF )   Comments on the Screening Results ( PDF )   Annual Research Report

Research Products

(32 results)

All 2022 2021 2020 2019 2018 Other

All Int'l Joint Research (6 results) Journal Article (13 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 13 results,  Open Access: 6 results) Presentation (11 results) (of which Int'l Joint Research: 10 results,  Invited: 11 results) Patent(Industrial Property Rights) (2 results) (of which Overseas: 1 results)

  • [Int'l Joint Research] University of Massachusetts/Thomas Jefferson University(米国)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] 上海科技大学(中国)

    • Related Report
      2021 Annual Research Report
  • [Int'l Joint Research] University of Massachusetts/Thomas Jefferson University(米国)

    • Related Report
      2020 Annual Research Report
  • [Int'l Joint Research] 上海科技大学(中国)

    • Related Report
      2020 Annual Research Report
  • [Int'l Joint Research] Thomas Jefferson University(米国)

    • Related Report
      2019 Annual Research Report
  • [Int'l Joint Research] Chinese Academy of Sciences(中国)

    • Related Report
      2019 Annual Research Report
  • [Journal Article] Life of RISC: Formation, action, and degradation of RNA-induced silencing complex2022

    • Author(s)
      Iwakawa Hiro-oki、Tomari Yukihide
    • Journal Title

      Molecular Cell

      Volume: 82 Pages: 30-43

    • DOI

      10.1016/j.molcel.2021.11.026

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Ribosome stalling caused by the Argonaute-microRNA-SGS3 complex regulates the production of secondary siRNAs in plants2021

    • Author(s)
      Iwakawa Hiro-oki、Lam Andy Y.W.、Mine Akira、Fujita Tomoya、Kiyokawa Kaori、Yoshikawa Manabu、Takeda Atsushi、Iwasaki Shintaro、Tomari Yukihide
    • Journal Title

      Cell Reports

      Volume: 35 Pages: 109300-109300

    • DOI

      10.1016/j.celrep.2021.109300

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single-molecule analysis of processive double-stranded RNA cleavage by Drosophila Dicer-22021

    • Author(s)
      Naganuma Masahiro、Tadakuma Hisashi、Tomari Yukihide
    • Journal Title

      Nature Communications

      Volume: 12 Pages: 4268-4268

    • DOI

      10.1038/s41467-021-24555-1

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] RNase κ promotes robust piRNA production by generating 2',3'-cyclic phosphate-containing precursors.2021

    • Author(s)
      Shigematsu M, Kawamura T, Morichika K, Izumi N, Kiuchi T, Honda S, Pliatsika V, Matsubara R, Rigoutsos I, Katsuma S, Tomari Y, Kirino Y.
    • Journal Title

      Nature Communications

      Volume: 12 Pages: 4498-4498

    • DOI

      10.1038/s41467-021-24681-w

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Cell-free reconstitution reveals the molecular mechanisms for the initiation of secondary siRNA biogenesis in plants2021

    • Author(s)
      Sakurai Yuriki、Baeg Kyungmin、Lam Andy Y. W.、Shoji Keisuke、Tomari Yukihide、Iwakawa Hiro-oki
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 118

    • DOI

      10.1073/pnas.2102889118

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Mechanistic analysis of the enhanced RNAi activity by 6-mCEPh-purine at the 5′ end of the siRNA guide strand2020

    • Author(s)
      Brechin Vincent、Shinohara Fumikazu、Saito Jun-ichi、Seitz Herv?、Tomari Yukihide
    • Journal Title

      RNA

      Volume: 27 Pages: 151-162

    • DOI

      10.1261/rna.073775.119

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] siRNA potency enhancement via chemical modifications of nucleotide bases at the 5′-end of the siRNA guide strand2020

    • Author(s)
      Shinohara Fumikazu、Oashi Taiji、Harumoto Toshimasa、Nishikawa Tomoyuki、Takayama Yuki、Miyagi Hikaru、Takahashi Yuichi、Nakajima Takahiro、Sawada Takashi、Koda Yasuo、Makino Asana、Sato Atsuko、Hamaguchi Kaori、Suzuki Michihiko、Yamamoto Junichiro、Tomari Yukihide、Saito Jun-Ichi
    • Journal Title

      RNA

      Volume: 27 Pages: 163-173

    • DOI

      10.1261/rna.073783.119

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Zucchini consensus motifs determine the mechanism of pre-piRNA production2020

    • Author(s)
      Izumi Natsuko、Shoji Keisuke、Suzuki Yutaka、Katsuma Susumu、Tomari Yukihide
    • Journal Title

      Nature

      Volume: 578 Pages: 311-316

    • DOI

      10.1038/s41586-020-1966-9

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A widespread family of heat-resistant obscure (Hero) proteins protect against protein instability and aggregation2020

    • Author(s)
      Tsuboyama K., Osaki T., Matsuura-Suzuki E., Kozuka-Hata H., Okada Y., Oyama M., Ikeuchi Y., Iwasaki S., and Tomari Y.
    • Journal Title

      PLoS Biol.

      Volume: 18 Issue: 3 Pages: 3000632-3000632

    • DOI

      10.1371/journal.pbio.3000632

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] VCP Machinery Mediates Autophagic Degradation of Empty Argonaute2019

    • Author(s)
      Kobayashi Hotaka、Shoji Keisuke、Kiyokawa Kaori、Negishi Lumi、Tomari Yukihide
    • Journal Title

      Cell Reports

      Volume: 28 Pages: 1144-1153

    • DOI

      10.1016/j.celrep.2019.07.003

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Identification of an AGO (Argonaute) protein as a prey of TER94/VCP2019

    • Author(s)
      Kobayashi Hotaka、Tomari Yukihide
    • Journal Title

      Autophagy

      Volume: 16 Pages: 190-192

    • DOI

      10.1080/15548627.2019.1691351

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Iruka Eliminates Dysfunctional Argonaute by Selective Ubiquitination of Its Empty State2019

    • Author(s)
      Kobayashi Hotaka, Shoji Keisuke, Kiyokawa Kaori, Negishi Lumi, Tomari Yukihide
    • Journal Title

      Mol Cell

      Volume: 73 Pages: 119-129

    • DOI

      10.1016/j.molcel.2018.10.033

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Conformational Activation of Argonaute by Distinct yet Coordinated Actions of the Hsp70 and Hsp90 Chaperone Systems2018

    • Author(s)
      Tsuboyama K, Tadakuma H, Tomari Y
    • Journal Title

      Molecular Cell

      Volume: 70 Pages: 722-729

    • DOI

      10.1016/j.molcel.2018.04.010

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Presentation] Assembly of the RNA Silencing Complex (and Beyond)2021

    • Author(s)
      泊 幸秀
    • Organizer
      2021 RNA Biology Symposium
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] RNAサイレンシング複合体の形成と機能2021

    • Author(s)
      泊 幸秀
    • Organizer
      日本エピジェネティクス研究会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] Hero proteins: widespread protectors against protein instability2020

    • Author(s)
      泊 幸秀
    • Organizer
      MBSJ 2020 Workshop
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] The mechanism of pre-piRNA production2020

    • Author(s)
      泊 幸秀
    • Organizer
      UTRs in Development and Disease
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Assembly of the RNA-induced silencing complex and beyond2019

    • Author(s)
      泊 幸秀
    • Organizer
      RiboClub the 20th Anniversary
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Assembly and function of the RNA silencing complex2019

    • Author(s)
      泊 幸秀
    • Organizer
      FASEB SRC The RNA Localization and Local Translation Conference
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 3′-end maturation of silkworm piRNAs2019

    • Author(s)
      泊 幸秀
    • Organizer
      Keystone Symposia Small Regulatory RNAs
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] A conserved step-wise maturation mechanism of piRNAs2018

    • Author(s)
      泊 幸秀
    • Organizer
      EMBO Workshop: piRNAs and PIWI proteins
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Biochemical and biophysical dissection of RNA silencing2018

    • Author(s)
      泊 幸秀
    • Organizer
      RNA-REG: Regulatory Circuits in RNA Biology
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Iruka ensures the quality of Argonaute by selective ubiquitination of its empty state2018

    • Author(s)
      泊 幸秀
    • Organizer
      CSH Asia Meeting on RNA Biology
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Iruka ensures the quality of Argonaute by selective ubiquitination of its empty state2018

    • Author(s)
      泊 幸秀
    • Organizer
      Joint Australia-Japan RNA Meeting 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Patent(Industrial Property Rights)] クライアントタンパク質を保護するタンパク質のスクリーニング方法並びに生理活性タンパク質安定化タンパク質および該タンパク質を含む医薬組成物2019

    • Inventor(s)
      泊 幸秀、岩崎 信太郎、坪山 幸太郎、他4名
    • Industrial Property Rights Holder
      東京大学
    • Industrial Property Rights Type
      特許
    • Filing Date
      2019
    • Related Report
      2019 Annual Research Report
  • [Patent(Industrial Property Rights)] Protein stabilization by fully unstructured proteins2018

    • Inventor(s)
      坪山 幸太郎, 岩崎 信太郎, 泊 幸秀
    • Industrial Property Rights Holder
      東京大学
    • Industrial Property Rights Type
      特許
    • Filing Date
      2018
    • Related Report
      2018 Annual Research Report
    • Overseas

URL: 

Published: 2018-06-12   Modified: 2022-12-28  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi