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Revalidation of the origin of DNA molecules by constructing Escherichia coli deficient in the core enzyme of RNA origin theory

Research Project

Project/Area Number 20K21274
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 38:Agricultural chemistry and related fields
Research InstitutionKyoto University

Principal Investigator

Ogawa Jun  京都大学, 農学研究科, 教授 (70281102)

Co-Investigator(Kenkyū-buntansha) 竹内 道樹  京都大学, 農学研究科, 特定助教 (40766193)
Project Period (FY) 2020-07-30 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2022: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2021: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2020: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywordsデオキシリボヌクレオシド / リボヌクレオチド還元酵素 / DNA / リボヌクレオシド還元酵素 / 遺伝子破壊
Outline of Research at the Start

DNA分子の起源をRNAとするRNA起源説には、いくつかの問題点が存在するものの、他に有力な仮説がないことから、広く受け入れられている。本研究では、RNA起源説に一石を投ずるべく、すべての生物に共通でRNAからのDNA合成の鍵酵素である「リボヌクレオチド還元酵素(RNR)」を欠損する大腸菌を構築する。さらに、「新規DNA合成経路の探索・機能検証」の成果を互いにフィードバックさせることで、生命分子起源における新たな仮説の提案を試みる。加えて、見いだされる新規dNS生合成経路を、dNSの新規製法デザインに展開する。

Outline of Final Research Achievements

We have succeeded in constructing an E. coli that lacks ribonucleotide reductase (RNR), a key enzyme for DNA synthesis from RNA that is common to all organisms. When the nutrient requirements of this bacteria were examined, it was observed that it requires deoxycytidine (dC) among deoxyribonucleosides (dNS), as well as several vitamins.
When a disruption strain expressing a gene for the deoxyribonucleoside synthesis pathway was evaluated for its ability to synthesize dNS, it successfully synthesized deoxyuridine but not dC.
Bioassays using dNS-requiring strains were used to search for novel deoxyribonucleoside-synthesizing microorganisms, but no strains with a novel pathway were found.

Academic Significance and Societal Importance of the Research Achievements

原始地球上にRNAが最初に存在したと仮定するRNAワールド仮説は、広く信じられている。地球上のすべての生物は、リボヌクレオチドの還元反応によりDNAの構成分子を酵素合成する。本反応を触媒する酵素がribonucleotide reductase(RNR)であり、RNA仮説における「DNAはRNAに由来する」とする説の中核を担っている。本来必須遺伝子とされるRNR遺伝子を破壊し、かつ、新たなdNS生合成系により太古の自然環境に存在しえた化合物からDNAを生合成し生育しうる生物を構築できれば、DNAがRNAに由来するというRNA起源説に一石を投じるとともに、新たな生命分子起源説を提案しうる。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • Research Products

    (7 results)

All 2023 2022 2021 Other

All Int'l Joint Research (2 results) Journal Article (1 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (4 results) (of which Int'l Joint Research: 1 results)

  • [Int'l Joint Research] The University of Auckland(ニュージーランド)

    • Related Report
      2022 Annual Research Report
  • [Int'l Joint Research] The University of Auckland(ニュージーランド)

    • Related Report
      2020 Research-status Report
  • [Journal Article] Characterisation of an Escherichia coli line that completely lacks ribonucleotide reduction yields insights into the evolution of obligate intracellularity.2023

    • Author(s)
      Arras, S. D. M., N. Sibaeva, R. J. Catchpole, N. Horinouchi, D. Si, A. M. Rickerby, K. Deguchi, M. Hibi, K. Tanaka, M. Takeuchi, J. Ogawa, A. M. Poole.
    • Journal Title

      eLife

      Volume: 12

    • DOI

      10.7554/elife.83845

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Evaluation of auxotrophy of Escherichia coli disrupted ribonucleotide reductase genes and functional verification of the novel dNS synthetic pathway2022

    • Author(s)
      Kengo Deguchi, Dayoung Si, Nobuyuki Horinouchi, Syoko Kozono, Michiki Takeuchi, Makoto Hibi, Anthony M. Poole, Jun Ogawa
    • Organizer
      Active Enzyme Molecule 2022
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] デオキシリボヌクレチド生合成経路遺伝子を破壊し代替経路を導入した大腸菌による デオキシリボヌクレオシド生産の検討2022

    • Author(s)
      出口賢児 、斯大勇、堀之内伸行、小園祥子 、竹内道樹、 日比慎 、Anthony M. Poole、小川順
    • Organizer
      酵素工学研究会
    • Related Report
      2021 Research-status Report
  • [Presentation] デオキシリボヌクレオチド生合成経路遺伝子を破壊した大腸菌の生育評価と代替経路導入の検証2022

    • Author(s)
      出口賢児 、斯大勇、堀之内伸行、小園祥子 、竹内道樹、 日比慎 、Anthony M. Poole、小川順
    • Organizer
      農芸化学会
    • Related Report
      2021 Research-status Report
  • [Presentation] Growth evaluation of Escherichia coli with knockout of deoxyribonucleotide synthetic pathway genes2021

    • Author(s)
      出口賢児, 斯大勇, 堀之内伸行, 小園祥子, 竹内道樹, 日比慎, Anthony M. poole, 小川順
    • Organizer
      日本農芸化学会2021年度大会
    • Related Report
      2020 Research-status Report

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Published: 2020-08-03   Modified: 2024-01-30  

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