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Genetic study on bacterial colony formation and its application to isolation of bacteria

Research Project

Project/Area Number 17H03789
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied microbiology
Research InstitutionTokyo University of Agriculture (2020-2021)
The University of Tokyo (2017-2019)

Principal Investigator

Masaki Haruhiko  東京農業大学, 生命科学部, 教授 (50134515)

Co-Investigator(Kenkyū-buntansha) 清水 則夫  東京医科歯科大学, 統合研究機構, 准教授 (30226245)
Project Period (FY) 2017-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000)
Fiscal Year 2020: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2019: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2018: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2017: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
Keywords大腸菌 / コロニー形成 / 温度感受性変異 / MPN法 / 液体培養 / 不飽和脂肪酸 / cAMP / VBNC / コリシン / tRNA / コロニー / 脂肪酸 / バクテリア / コリシンD / 硫黄代謝 / 脂肪酸合成 / CRP / 細菌 / 増殖 / cAMP受容タンパク質 / ASKA
Outline of Final Research Achievements

Most of bacteria in nature do not form colonies on solid media. To know its genetic background, we isolated Escherichia coli mutants that hardly form colonies compared to liquid cultivation. Characterization of the mutants revealed the importance of unsaturated fatty acids in colony formation. When faced with starvation, even wild-type E. coli falls in shortage of fatty acids, causing decrease of colony formation. When fatty acid synthesis was inhibited with chemical drugs, colony formation was impaired in not only E. coli but also Bacillus subtilis, Staphylococcal epidermidis and Corynebacterium glutamicum, suggesting its generality in bacteria.
In the course of screening of essential genes for the maintenance of colony-forming activity of E. coli in the viable but non-culturable state, we found that both cyaA- and & crp- strains keep high colony-forming activities for a long time, suggesting that cAMP-CRP is a global switch to lead cells to dormancy on starvation stress.

Academic Significance and Societal Importance of the Research Achievements

伝統的に細菌が生きている証拠とされてきたコロニー形成を相対化し,コロニーを作れない変異体を分離することで,コロニー形成が特殊な生物現象であることを示した。その変異を糸口にして,飢餓に接した細菌がコロニー形成能を落とす隠れた一因が,栄養要求性とは異なる不飽和脂肪酸の欠乏にあることを見出し,その供給が一般細菌の分離にも重要であることを示唆した。
飢餓に曝された大腸菌がコロニーを作らなくなるVBNC現象を,細胞増殖能の減衰という一面だけでなく, cAMPをシグナルとして増殖能を積極的に遮断して休眠状態に入っていく過程である可能性を示した。

Report

(5 results)
  • 2021 Final Research Report ( PDF )
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (10 results)

All 2021 2020 2019 2018 2017 Other

All Journal Article (6 results) (of which Peer Reviewed: 4 results,  Open Access: 3 results) Presentation (3 results) (of which Invited: 1 results) Remarks (1 results)

  • [Journal Article] Substrate recognition mechanism of tRNA-targeting ribonuclease, colicin D, and an insight into tRNA cleavage-mediated translation impairment.2021

    • Author(s)
      Ogawa, T., Takahashi, K., Ishida, W., Aono, T., Hidaka, M., Terada, T., Masaki, H.
    • Journal Title

      RNA Biology

      Volume: 18 Issue: 8 Pages: 1193-1205

    • DOI

      10.1080/15476286.2020.1838782

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] バクテリアのコロニー形成の遺伝学事始め2019

    • Author(s)
      正木春彦,納庄一樹
    • Journal Title

      バイオサイエンスとインダストリー

      Volume: 77 Pages: 221-225

    • NAID

      40021932037

    • Related Report
      2019 Annual Research Report
  • [Journal Article] コロニー形成の遺伝学,その後2019

    • Author(s)
      正木春彦
    • Journal Title

      IFO Res. Commun.

      Volume: 33 Pages: 221-222

    • Related Report
      2019 Annual Research Report
  • [Journal Article] Isolation of colonization-defective Escherichia coli mutants reveals critical requirement for fatty acids in bacterial colony formation.2018

    • Author(s)
      Kazuki Nosho, Koji Yasuhara, Yuto Ikehata, Tomohiro Mii, Taichiro Ishige, Shunsuke Yajima, Makoto Hidaka, Tetsuhiro Ogawa, Haruhiko Masaki
    • Journal Title

      Microbiology

      Volume: 164 Issue: 9 Pages: 1122-1132

    • DOI

      10.1099/mic.0.000673

    • Related Report
      2018 Annual Research Report 2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Crystal structure of the central and the C-terminal RNase domains of colicin D implicated its translocation pathway through inner membrane of target cell2018

    • Author(s)
      Chang Jung-Wei、Sato Yusuke、Ogawa Tetsuhiro、Arakawa Takatoshi、Fukai Shuya、Fushinobu Shinya、Masaki Haruhiko
    • Journal Title

      The Journal of Biochemistry

      Volume: 164 Issue: 5 Pages: 329-339

    • DOI

      10.1093/jb/mvy056

    • NAID

      40021724807

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Linear Regression Links Transcriptomic Data and Cellular Raman Spectra2018

    • Author(s)
      Kobayashi-Kirschvink, K. J., Nakaoka, H., Oda, A., Kamei, K. F., Nosho, K., Fukushima, H., Kanesaki, Y., Yajima, S., Masaki, H., Ohta, K., Wakamoto, Y.
    • Journal Title

      Cell Systems

      Volume: 7 Issue: 1 Pages: 104-117

    • DOI

      10.1016/j.cels.2018.05.015

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] バクテリアが飢餓環境中でコロニーを作らなくなるのはなぜか?2020

    • Author(s)
      正木春彦
    • Organizer
      第35回日本環境感染学会総会・学術集会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] 脂肪酸の供給不足がバクテリアのコロニー形成能を失わせる: コロニーを作らない大腸菌変異株の性質からの一般化2018

    • Author(s)
      納庄一樹, 三井智玄, 池端佑仁, 安原幸司, 正木春彦
    • Organizer
      第17回 微生物研究会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Molecular structure of colicin D, an Escherichia coli toxin that cleaves tRNAs for arginine2017

    • Author(s)
      Jung-Wei Chang, Yusuke Sato, Tetsuhiro Ogawa, Takatoshi Arakawa1 Shinya Fushinobu, Shuya Fukai, Haruhiko Masaki
    • Organizer
      第17回 日本蛋白質科学会年会
    • Related Report
      2017 Annual Research Report
  • [Remarks] コロニーを作れない大腸菌が教える: 自然界のバクテリアは脂肪酸欠乏に陥ってコロニーが作れなくなる

    • URL

      https://www.a.u-tokyo.ac.jp/topics/2018/20181001-1.html

    • Related Report
      2018 Annual Research Report

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Published: 2017-04-28   Modified: 2023-01-30  

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