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Analysis of the molecular mechanisms of RNA-directed DNA methylation

Research Project

Project/Area Number 16H06159
Research Category

Grant-in-Aid for Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field Molecular biology
Research InstitutionThe University of Tokyo

Principal Investigator

Iwakawa Hiro-oki  東京大学, 定量生命科学研究所, 助教 (60710415)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥24,310,000 (Direct Cost: ¥18,700,000、Indirect Cost: ¥5,610,000)
Fiscal Year 2018: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
Fiscal Year 2017: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
Fiscal Year 2016: ¥10,660,000 (Direct Cost: ¥8,200,000、Indirect Cost: ¥2,460,000)
KeywordsRNAサイレンシング / siRNA / DNA-RNAハイブリッド / RNA-DNAハイブリッド / RNA-DNA ハイブリッド
Outline of Final Research Achievements

We conducted our research by focusing on how RNA-induced silencing complexes (RISCs) mediate sequence specific DNA methylation in nuclei. Through in vitro binding assays, we found that the plant cytoplasmic RISCs interact more tightly with RNAs than with DNAs, conversely, the plant nuclear RISCs bind to DNAs with much higher affinity than to RNAs. Moreover, we obtained novel insights into nuclear RISC assembly and template specificity of RNA-dependent RNA polymerase 6 (RDR6), which is known to be involved in de novo DNA methylation.

Academic Significance and Societal Importance of the Research Achievements

植物の核内RISCがRNAよりもDNAと強く結合することを世界で初めて生化学的に示したことで、「核内RISCは新生RNAとの結合を介して標的DNAに接近しメチル化を促進する」と信じられてきた従来のモデルを書き換える可能性がある。さらに核内RISC形成機構やRDR6の鋳型特異性の生化学的な理解は、未だ多くの謎に包まれている核内RNAサイレンシング機構を理解する上で重要な基盤的知見となる。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Annual Research Report
  • 2016 Annual Research Report
  • Research Products

    (20 results)

All 2019 2018 2017 2016 Other

All Journal Article (5 results) (of which Peer Reviewed: 5 results,  Open Access: 2 results,  Acknowledgement Compliant: 1 results) Presentation (13 results) (of which Int'l Joint Research: 7 results,  Invited: 3 results) Remarks (2 results)

  • [Journal Article] In vitro RNA-dependent RNA Polymerase Assay Using Arabidopsis RDR62018

    • Author(s)
      Kyungmin Baeg, Yukihide Tomari, Hiro-oki Iwakawa
    • Journal Title

      Bio-protocol

      Volume: 8 Issue: 1

    • DOI

      10.21769/bioprotoc.2673

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] ilencing messages in a unique way2017

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

      Nature Plants,

      Volume: 3 Issue: 10 Pages: 769-770

    • DOI

      10.1038/s41477-017-0028-2

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] In Vitro Analysis of ARGONAUTE-Mediated Target Cleavage and Translational Repression in Plants2017

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

      Methods in Molecular Biology

      Volume: 1640 Pages: 55-71

    • DOI

      10.1007/978-1-4939-7165-7_4

    • ISBN
      9781493971640, 9781493971657
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 植物のRNAサイレンシング機構が自己のmRNAを攻撃しないメカニズム2017

    • Author(s)
      岩川 弘宙
    • Journal Title

      BSJ-Review

      Volume: 8 Issue: B Pages: 58-70

    • DOI

      10.24480/bsj-review.8b4.00114

    • ISSN
      2432-9819
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] The poly(A) tail blocks RDR6 from converting self mRNAs into substrates for gene silencing.2017

    • Author(s)
      Baeg K, *Iwakawa HO, *Tomari Y.
    • Journal Title

      Nature Plants

      Volume: 3 Issue: 4 Pages: 17036-17036

    • DOI

      10.1038/nplants.2017.36

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] In vitro recapitulation of the secondary siRNA biogenesis in plants2019

    • Author(s)
      Hiro-oki Iwakawa, Kyungmin Baeg, Yuriki Sakurai, Manabu Yoshikawa, Yukihide Tomari
    • Organizer
      第20回武田科学振興財団生命科学シンポジウム RNAネオバイオロジー
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] In vitro recapitulation of the secondary siRNA biogenesis in plants2019

    • Author(s)
      Kyungmin Baeg, Yuriki Sakurai, Andy Lam, Keisuke Shoji, Manabu Yoshikawa, Yukihide Tomari, Hiro-oki Iwakawa
    • Organizer
      POST-TRANSCRIPTIONAL GENE REGULATION IN PLANTS (PGRP) 2019 Nara
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] The PIWI domain of RdDM Argonautes in plants plays a critical role for sensing the identity of the 5′-base of small RNAs2019

    • Author(s)
      Wei Liu, Yukihide Tomari, Hiro-oki Iwakawa
    • Organizer
      POST-TRANSCRIPTIONAL GENE REGULATION IN PLANTS (PGRP) 2019 Nara
    • Related Report
      2018 Annual Research Report
  • [Presentation] 植物のRNAサイレンシング機構2018

    • Author(s)
      岩川弘宙
    • Organizer
      第53回 植物感染生理談話会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 植物における二次的小分子RNA生成の試験管内再現2018

    • Author(s)
      Baeg Kyungmin、櫻井 友理希、吉川 学、泊 幸秀、岩川 弘宙
    • Organizer
      第20回日本RNA学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] In vitro analysis of small RNA preference of ARGONAUTE 4 in Arabidopsis thaliana2018

    • Author(s)
      Wei Liu, Yukihide Tomari, Hiro-oki Iwakawa
    • Organizer
      第59回日本植物生理学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Plant ARGONAUTE4 family proteins prefer to bind to DNA targets in vitro2017

    • Author(s)
      Iwakawa HO, Tomari Y
    • Organizer
      The 22nd Annual Meeting of the RNA Society
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] The poly(A) tail blocks RDR6 from converting self mRNAs into the substrates for gene silencing2017

    • Author(s)
      Baeg K, Iwakawa HO, Tomari Y
    • Organizer
      The 22nd Annual Meeting of the RNA Society
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Plant ARGONAUTE4 family proteins prefer to bind to DNA targets in vitro2017

    • Author(s)
      Iwakawa HO, Tomari Y
    • Organizer
      12th Microsymposium on small RNA biology
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] The poly(A) tail blocks RDR6 from converting self mRNAs into the substrates for gene silencing2017

    • Author(s)
      Baeg K, Iwakawa HO, Tomari Y
    • Organizer
      12th Microsymposium on small RNA biology
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Plant ARGONAUTE4 family proteins prefer to bind to DNA targets in vitro2016

    • Author(s)
      Hiro-oki Iwakawa, Yukihide Tomari
    • Organizer
      The 2016 joint annual meeting of the RNA Society and the RNA Society of Japan
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 植物のRNAサイレンシング機構2016

    • Author(s)
      岩川 弘宙
    • Organizer
      第39回日本分子生物学会
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] 生化学で迫る植物の RNA サイレンシング2016

    • Author(s)
      岩川 弘宙
    • Organizer
      日本植物学会第80回大会
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Remarks] researchmap

    • URL

      https://researchmap.jp/Hiro_Iwa/

    • Related Report
      2018 Annual Research Report
  • [Remarks] researchmap 岩川 弘宙

    • URL

      https://researchmap.jp/Hiro_Iwa/

    • Related Report
      2017 Annual Research Report 2016 Annual Research Report

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Published: 2016-04-21   Modified: 2020-03-30  

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