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Visualization of the RIP machinery in living Neurospora

Research Project

Project/Area Number 15K14916
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Applied molecular and cellular biology
Research InstitutionUniversity of Fukui

Principal Investigator

Honda Shinji  福井大学, 学術研究院医学系部門, 助教 (90632167)

Project Period (FY) 2015-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2015: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Keywordsゲノム防御 / ゲノム編集 / 染色体動態 / ライブイメージング / アカパンカビ
Outline of Final Research Achievements

The filamentous fungi Neurospora crassa has a unique genome defense system called Repeat Induced Point mutation (RIP). During specific sexual phase, RIP detects duplicated sequences throughout genome and mutates them, ultimately resulting in gene disruption. The mechanism, however, remains almost unknown. This study aimed to label artificial duplicated sequences by fluorescent proteins and to trace the RIP machinery in living cells. We first optimized blue, green, red, and near-infrared fluorescent protein tagging systems and successfully visualized distinct nuclear compartments in Neurospora. Furthermore, we established a method to effectively produce transparent perithecia to enhance the detection of fluorescent proteins in sexual phases. We are currently optimizing the visualization of artificial duplicated sequences tethered by our improved fluorescent protein systems.

Report

(3 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • Research Products

    (6 results)

All 2016 Other

All Journal Article (3 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 3 results) Presentation (2 results) (of which Int'l Joint Research: 2 results) Remarks (1 results)

  • [Journal Article] Normal chromosome conformation depends on subtelomeric facultative heterochromatin Neurospora crassa2016

    • Author(s)
      Klocko AD, Ormsby T, Galazka JM, Leggett N, Uesaka M, Honda S, Freitag M, and Selker EU
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 113 Issue: 52 Pages: 15048-15053

    • DOI

      10.1073/pnas.1615546113

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Dual chromatin recognition by the histone deacetylase complex HCHC is required for proper DNA methylation in Neurospora crassa2016

    • Author(s)
      Honda S, Bicocca VT, Gessaman JD, Rountree MR, Yokoyama A, Yu EY, Selker JM, and Selker EU
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 113 Issue: 41

    • DOI

      10.1073/pnas.1614279113

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Neurospora chromosomes are organized by blocks of importin alpha-dependent heterochromatin that are largely independent of H3K9me32016

    • Author(s)
      Galazka JM, Klocko AD, Uesaka M, Honda S, Selker EU, Freitag M
    • Journal Title

      Genome Research

      Volume: 26 Issue: 8 Pages: 1069-1080

    • DOI

      10.1101/gr.203182.115

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] Shelterin is required for telomeric integrity in Neurospora crassa2016

    • Author(s)
      Uesaka M, Yokoyama A, Lewis ZA and Honda S
    • Organizer
      EMBO conference
    • Place of Presentation
      Liege, Belgium
    • Year and Date
      2016-04-26
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Presentation] Shelterin is required for telomeric integrity in Neurospora crassa2016

    • Author(s)
      Uesaka M, Yokoyama A, Lewis ZA and Honda S
    • Organizer
      Neurospora meeting 2016
    • Place of Presentation
      California, USA
    • Year and Date
      2016-03-10
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Remarks] 福井大学 学術研究院医学系部門 生命情報医科学講座 染色体機能学領域

    • URL

      http://chromosome.med.lab.u-fukui.ac.jp

    • Related Report
      2016 Annual Research Report

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Published: 2015-04-16   Modified: 2018-03-22  

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