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Spatiotemporal mapping of gene expression to reveal mechanisms of cellular differentiation in E. coli biofilms

Research Project

Project/Area Number 21K05341
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 38020:Applied microbiology-related
Research InstitutionKyoto University

Principal Investigator

ROBERT Martin  京都大学, 薬学研究科, 特定准教授 (90365487)

Project Period (FY) 2021-04-01 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2023: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2022: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2021: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywordsbiofilm / E. coli / gene expression / imaging / differentation / bacterial growth / proteomics / stress resistance / proteome analysis / cell motility / acid fermentation / imaging system / proteome / surface properties / nutrients / differentiation / Escherichia coli
Outline of Research at the Start

Bacteria can form biofilms having unique and emergent biological properties. To explore these properties we will quantify gene expression and metabolic activity in E. coli biofilms using time-lapse fluorescence imaging of gene expression, and spatio-temporal mapping of the cellular proteome.

Outline of Final Research Achievements

This project was about examining functional differentiation in E. coli biofilms by monitoring growth, morphology and gene expression during development.
We found that in complex media glucose affects biofilm formation resulting in structural differences as well as functional differences linked to metabolism and response to acid stress during growth on glucose. Differential spatio-temporal expression was observed both with fluorescent reporters and proteome analysis. Morphological differences arise from changes in surface hardness as well as local nutritional conditions. We observed changes in gene expression activity and protein levels related to motility, acid resistance, metabolism, biofilm formation and protein synthesis. As planned, we provided novel evidence for spatiotemporal differentiation in activity in different parts of the biofilm (top-bottom and center-edge axis) and over time (biofilm growth and aging) that suggest co-existence and interactions between these populations.

Academic Significance and Societal Importance of the Research Achievements

この結果はバイオフィルムの成長と発達に対するグルコースの重要な影響を浮き彫りにした。 また、環境条件や薬剤などに対する耐性を試験するために使用できる単純な生物学的モデルにおける、多細胞の増殖と分化に関する基本的な洞察を提供する。バイオフィルムの生存と形成に重要な耐酸性、マトリックス産生、運動性遺伝子に関連した明らかな変化は、バイオフィルムの形成を制御するための新たなターゲットとなる可能性を示唆している。加えて、我々は関連分野の他の研究者にも有用な、オープンソースタイプの汎用的で安価なマルチモードイメージングシステムを開発した。

Report

(4 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Research-status Report
  • 2021 Research-status Report
  • Research Products

    (13 results)

All 2023 2022 2021 Other

All Presentation (12 results) (of which Int'l Joint Research: 2 results,  Invited: 1 results) Remarks (1 results)

  • [Presentation] Exploring culture conditions and fractionation methods for spatio-temporal analysis of E. coli biofilms.2023

    • Author(s)
      Fujino. Y and Robert, M.
    • Organizer
      The 46th Annual Meeting of the Molecular Biology Society of Japan
    • Related Report
      2023 Annual Research Report
  • [Presentation] Investigating the role of YccA on E. coli biofilm growth and development.2023

    • Author(s)
      Zhao, Y and Robert, M.
    • Organizer
      The 46th Annual Meeting of the Molecular Biology Society of Japan
    • Related Report
      2023 Annual Research Report
  • [Presentation] Development of imaging system to track biofilm morphology and temporal gene expression in growing biofilms.2023

    • Author(s)
      Sakai. R, Zhao, Y, and Robert, M.
    • Organizer
      The 46th Annual Meeting of the Molecular Biology Society of Japan
    • Related Report
      2023 Annual Research Report
  • [Presentation] Biofilming beyond B. subtilis2023

    • Author(s)
      Robert, M.
    • Organizer
      BACELL 2023
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research
  • [Presentation] E. coli biofilms as a developmental model2023

    • Author(s)
      Robert M.
    • Organizer
      第19回 21世紀大腸菌研究会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Effects of glucose on the growth of E. coli biofilms in rich medium2023

    • Author(s)
      干場 悠介, 藤野 祐紀子, Zhao Yifan, 酒井 隆之介, Robert M.
    • Organizer
      第19回 21世紀大腸菌研究会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Investigating the role of YccA on E. coli biofilm growth and development2023

    • Author(s)
      Zhao Yifan, Sakai R., Fujino Y., Hoshiba Y., Robert M.
    • Organizer
      第19回 21世紀大腸菌研究会
    • Related Report
      2023 Annual Research Report
  • [Presentation] 明視野および蛍光モードでバイオフィルムの動態を捉える タイムラプスイメージングシステムの開発2023

    • Author(s)
      酒井 隆之介, Zhao Yifan, Robert M
    • Organizer
      第19回 21世紀大腸菌研究会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Exploring patterns of growth and differentiation during E. coli biofilm formation.2023

    • Author(s)
      Robert M.
    • Organizer
      日本農芸化学会関西支部 第525回講演会
    • Related Report
      2023 Annual Research Report
  • [Presentation] Exploring transitions in metabolic state during the development of bacterial biofilms.2023

    • Author(s)
      Fujino Y. Sakai, R. and Robert M.
    • Organizer
      RIKEN BDR Symposium 2023 “Transitions in Biological Systems”. Kobe
    • Related Report
      2022 Research-status Report
    • Int'l Joint Research
  • [Presentation] Opening and closing the box of bacterial metabolic function: from enzyme discovery to adaptive evolution and biofilms using E. coli.2022

    • Author(s)
      Robert M.
    • Organizer
      Annual Meeting of Japan Society for Environmental Biotechnology. Tokyo
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] Exploring developmental processes in growing E. coli biofilms2021

    • Author(s)
      Yukiko Fujino
    • Organizer
      ゲノム微生物学会
    • Related Report
      2021 Research-status Report
  • [Remarks] Biofilm spatio-temporal gene expression mapping

    • URL

      https://sites.google.com/kyoto-u.ac.jp/msb/research/biofilm-spatiotemporal-mapping-project

    • Related Report
      2023 Annual Research Report

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Published: 2021-04-28   Modified: 2025-01-30  

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