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Elucidation of the molecular mechanism of action of the yeast strongest DIT1 terminator and its application

Research Project

Project/Area Number 15K07378
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Applied microbiology
Research InstitutionToyota Central R&D Lab., Inc.

Principal Investigator

Matsuyama Takashi  株式会社豊田中央研究所, 社会システム研究領域 健康創出プログラム, 主任研究員 (90394882)

Co-Investigator(Kenkyū-buntansha) 井沢 真吾  京都工芸繊維大学, 応用生物学系, 准教授 (10273517)
北川 孝雄  山口大学, 大学院医学系研究科, 助教(特命) (20614928)
Co-Investigator(Renkei-kenkyūsha) IRIE Kenji  筑波大学, 医学医療系, 教授 (90232628)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords転写後発現制御 / ターミネーター / 出芽酵母 / 遺伝子発現 / RNA結合タンパク質
Outline of Final Research Achievements

In budding yeast, we analyzed the activation mechanism of the DIT1 terminator (DIT1t) with the highest activity to increase protein production of its upstream ORF. The cis sequences GUUCG/U were identified. Due to accumulation of deletion mutations that enhance the activity of DIT1t activation, an artificial DIT1t with about 1.5 times the activity of the wild type DIT1t was constructed.
Terminator activity of six genes involved in cell wall synthesis with GUUCG/U in the 3'-UTR was increased by overexpression of NAB6 and PAP1 genes. On the other hand, the nab6-disrupted strain was resistant to a cell wall synthetic inhibitor, caspofungin, and was sensitive to the drug congo red, which binds to another cell wall component chitin and causes growth inhibition. Taken together, it was suggested that a global regulation system using Nab6p-Pap1p complex as a hub might control cell wall synthesis.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (12 results)

All 2018 2017 2016 2015 Other

All Journal Article (2 results) (of which Peer Reviewed: 2 results,  Open Access: 2 results,  Acknowledgement Compliant: 1 results) Presentation (5 results) (of which Int'l Joint Research: 2 results) Book (1 results) Remarks (2 results) Patent(Industrial Property Rights) (2 results)

  • [Journal Article] Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae.2016

    • Author(s)
      Y. Ito, T. Kitagawa, M. Yamanashi, S. Katahira, S. Izawa, K. Irie, M. Furutani-Seiki, and T. Matsuyama
    • Journal Title

      Sci. Rep.

      Volume: 6 Issue: 1 Pages: 36997-36997

    • DOI

      10.1038/srep36997

    • NAID

      120007129256

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Combinatorial Screening for Transgenic Yeasts with High Cellulase Activities in Combination with a Tunable Expression System2015

    • Author(s)
      Yoichiro Ito, Mamoru Yamanishi, Akinori Ikeuchi, Chie Imamura, Takashi Matsuyama
    • Journal Title

      Plos One

      Volume: 10 Issue: 12 Pages: e0144870-e0144870

    • DOI

      10.1371/journal.pone.0144870

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] DIT1ターミネーターと2つのRNA結合タンパク質による目的タンパク質の増産技術2018

    • Author(s)
      松山崇、伊藤洋一郎、北川孝雄、山西守、片平悟史、井沢真吾、入江賢児、清木誠
    • Organizer
      日本農芸化学会2018年度大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae2017

    • Author(s)
      松山崇
    • Organizer
      The 33rd International Specialised Symposium on Yeast (ISSY33)
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] DIT1ターミネーターと2つのRNA結合タンパク質による目的タンパク質の増産技術2017

    • Author(s)
      松山崇、伊藤洋一郎、北川孝雄、山西守、片平悟史、井沢真吾、入江賢児、清木誠
    • Organizer
      日本生物工学会第69回大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Combinatorial screening for transgenic yeasts with high cellulase activities in combination with a highly tunable expression system2016

    • Author(s)
      松山崇
    • Organizer
      14th International Congress on Yeasts
    • Place of Presentation
      淡路島夢舞台(兵庫)
    • Year and Date
      2016-09-11
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] 細胞壁合成阻害剤を用いたNAB6遺伝子の機能解析2016

    • Author(s)
      松山崇、北川孝雄、入江賢児
    • Organizer
      日本農芸化学会 2016年度大会
    • Place of Presentation
      札幌コンベンションセンター、札幌市、北海道
    • Year and Date
      2016-03-29
    • Related Report
      2015 Research-status Report
  • [Book] 酵母菌・麹菌・乳酸菌の産業応用展開 第四章 酵母による高活性ターミネーターを利用したタンパク質高生産2018

    • Author(s)
      松山崇
    • Total Pages
      7
    • Publisher
      シーエムシー出版
    • ISBN
      9784781313177
    • Related Report
      2017 Annual Research Report
  • [Remarks] 山口大学ホームページ

    • URL

      http://www.yamaguchi-u.ac.jp/weeklynews/2016/_5595.html

    • Related Report
      2016 Research-status Report
  • [Remarks] 京都工芸繊維大学ホームページ

    • URL

      https://www.kit.ac.jp/2016/11/161117_releasenews/

    • Related Report
      2016 Research-status Report
  • [Patent(Industrial Property Rights)] 酵母ターミネーター及びその利用2016

    • Inventor(s)
      松山崇
    • Industrial Property Rights Holder
      株式会社豊田中央研究所
    • Industrial Property Rights Type
      特許
    • Filing Date
      2016
    • Acquisition Date
      2017
    • Related Report
      2017 Annual Research Report
  • [Patent(Industrial Property Rights)] 酵母ターミネーター及びその利用2016

    • Inventor(s)
      松山崇
    • Industrial Property Rights Holder
      株式会社 豊田中央研究所
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2016-003052
    • Filing Date
      2016-01-08
    • Related Report
      2015 Research-status Report

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Published: 2015-04-16   Modified: 2019-03-29  

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