Studies on formation mechanism of nano-organelle in growing cells by live-cell time-lapse fluorescence imaging analyses.

• Project/Area Number: 17H03791
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Kanazawa University
• Principal Investigator: Fukumori Yoshihirfo 金沢大学, その他部局等, その他 (60135655)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥15,730,000 (Direct Cost: ¥12,100,000、Indirect Cost: ¥3,630,000); Fiscal Year 2019: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2018: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000); Fiscal Year 2017: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
• Keywords: 磁性細菌 / オルガネラ / 原核細胞 / 生細胞イメージング / オルガネラ形成 / 細胞骨格 / 原子間力顕微鏡 / イメージング / 磁気微粒子 / ライブ解析 / ナノ

Outline of Final Research Achievements

The bacterial organelle "magnetosome" allows a bacterial cell to be sensing the magnetic field. In this study, we focused on the four magnetosomal proteins involved in the magnetosome vesicle formation process. We analyzed subcellular localizations of these vesicle forming proteins and identified protein partners interacting with these proteins in magnetosomes. The results suggested that the vesicle forming proteins localize in magnetosomes and interact with each other to form protein complexes. Moreover, these proteins interact with the magnetosomal proteins associated with iron uptake and redox reactions. Furthermore, we succeeded in observing the dynamic magnetosome assembly process by live-cell fluorescence imaging using the strain capable of genetically controlling magnetosome synthesis.

Academic Significance and Societal Importance of the Research Achievements

本研究では、「マグネトソーム」形成に必須の蛋白質の実態に世界で初めて迫った。これまでのところ、マグネトソームをはじめとする細菌の細胞内小器官の形成メカニズムは、ほとんど明らかにされていない。細菌の細胞内小器官は、CO2固定、窒素循環、水素生産、エネルギー生産などの生命活動と地球環境にとって重要な化学反応の場であり、その形成に必須の蛋白質に関する知見は、これらの反応の効率化や、新しい機能を有した細胞内小器官の創成を目指した研究の基礎として重要である。

Research Products

• [Journal Article] Tree of motility : A proposed history of motility systems in the tree of life (2020)
  • Author(s): M Miyata, R C Robinson, T QP Uyeda, Y Fukumori, S Fukushima, S Haruta, M Homma, K Inaba, M Ito, C Kaito, K Kato, TKenri, Y Kinosita, S Kojima, T Minamino, H Mori, S Nakamura, D Nakane, K Nakayama, M Nishiyama, S Shibata, K Shimabukuro, M Tamakoshi, A Taoka, Y Tashiro, ITulum, HiWada, K Wakabayashi
  • Journal Title: Genes to Cells Volume: 25 Issue: 1 Pages: 6-21
  • DOI: 10.1111/gtc.12737
  • NAID: 130007881664
  • URL: https://ocu-omu.repo.nii.ac.jp/records/2019864
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Diversity of physical properties of bacterial extracellular membrane vesicles revealed through atomic force microscopy phase imaging (2020)
  • Author(s): Kikuchi Yousuke、Obana Nozomu、Toyofuku Masanori、Kodera Noriyuki、Soma Takamitsu、Ando Toshio、Fukumori Yoshihiro、Nomura Nobuhiko、Taoka Azuma
  • Journal Title: Nanoscale Volume: 12 Issue: 14 Pages: 7950-7959
  • DOI: 10.1039/c9nr10850e
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Membranolytic Effects of KT2 on Gram-Negative Escherichia coli Evaluated by Atomic Force Microscopy (2019)
  • Author(s): Theansungnoen, T., Jangpromma, N., Anwised, P., Daduang, S., Fukumori, Y., Taoka, A., and Klaynongsruang
  • Journal Title: Applied Biochemistry and Microbiology Volume: 55(5) Issue: 5 Pages: 495-505
  • DOI: 10.1134/s0003683819050144
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Measuring magnetosomal pH of the magnetotactic bacterium Magnetospirillum magneticum AMB-1 using pH-sensitive fluorescent proteins (2018)
  • Author(s): Y. Eguchi, Y. Fukumori, A. Taoka
  • Journal Title: Biosci. Biotechnol. Biochem. Volume: in press Issue: 7 Pages: 1243-1251
  • DOI: 10.1080/09168451.2018.1451739
  • Peer Reviewed / Open Access
• [Journal Article] 細菌の磁気感応運動のためのオルガネラ「マグネトソーム」 (2018)
  • Author(s): 田岡東, 福森義宏
  • Journal Title: 生物工学 Volume: 96(5) Pages: 248-252
  • Peer Reviewed / Open Access
• [Journal Article] Role of the Bacterial Actin-like Cytoskeleton to Tethering Magnetic Organelle (2018)
  • Author(s): 田岡 東、福森義宏
  • Journal Title: Seibutsu Butsuri Volume: 58 Issue: 2 Pages: 091-093
  • DOI: 10.2142/biophys.58.091
  • NAID: 130006594317
  • ISSN: 0582-4052, 1347-4219
  • Peer Reviewed
• [Journal Article] Tethered Magnets Are the Key to Magnetotaxis: Direct Observations of Magnetospirillum magneticum AMB-1 Show that MamK Distributes Magnetosome Organelles Equally to Daughter Cells (2017)
  • Author(s): Taoka Azuma、Kiyokawa Ayako、Uesugi Chika、Kikuchi Yousuke、Oestreicher Zachery、Morii Kaori、Eguchi Yukako、Fukumori Yoshihiro
  • Journal Title: mBio Volume: 8 Issue: 4
  • DOI: 10.1128/mbio.00679-17
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Analysis of pH regulation in magnetotactic bacteria using pH-sensitive fluorescent protein (2020)
  • Author(s): 江口 友佳子, 田岡 東, 福森 義宏
  • Organizer: 第93回日本細菌学会
• [Presentation] maging of Dynamic Polymerization of Actin-Like Protein MamK for Bacterial Organelle Magnetosome Positioning. (2019)
  • Author(s): Taoka, A., Kikuchi, Y., and Fukumori Y.
  • Organizer: ASM microbe 2019 (米国微生物学会大会).
  • Int'l Joint Research
• [Presentation] irect imaging of membrane vesicle-mediated cell-to-cell communication using high-speed AFM (2019)
  • Author(s): 菊池 洋輔, 市中 佑樹, 豊福 雅典, 尾花 望, 古寺 哲幸, 安藤 敏夫, 福森 義宏, 野村 暢彦, 田岡 東
  • Organizer: 第92回 日本細菌学会総会
• [Presentation] pH感受性蛍光蛋白質を用いた原核細胞およびオルガネラ内のpH蛍光イメージング解析 (2018)
  • Author(s): 江口 友佳子 、田岡 東 、福森 義宏
  • Organizer: 第91回日本生化学会大会
• [Presentation] Measuring magnetosomal pH in living magnetotactic bacterial cell (2018)
  • Author(s): A. Taoka, Y. Eguchi, Y. Kikuchi, Y. Fukumori
  • Organizer: he 6th International Meeting on Magnetotactic Bacteria (MTB 2018 meeting)
  • Int'l Joint Research / Invited
• [Presentation] Structure and function of molecular machinery for magnetotaxis (2018)
  • Author(s): Y. Fukumori, A. Taoka
  • Organizer: The 6th International Meeting on Magnetotactic Bacteria (MTB 2018 meeting)
  • Int'l Joint Research / Invited
• [Presentation] 高速AFM解析により明らかになったグラム陰性菌及びグラム陽性菌の産生するメンブランベシクルの物性多様性 (2018)
  • Author(s): 菊池 洋輔, 尾花 望, 豊福 雅典, 野村 暢彦, 古寺 哲幸, 安藤 敏夫, 福森 義宏, 田岡 東
  • Organizer: 第56回日本生物物理学会年会
• [Presentation] High-speed atomic force microscopic observations explore molecule mechanism of bacterial cell-cell communication using membrane vesicles (2018)
  • Author(s): Yousuke Kikuchi, Nozomu Obana, Masanori Toyofuku, Nobuhiko Nomura, Noriyuki Kodera, Toshio Ando, Yoshihiro Fukumori, Azuma Taoka
  • Organizer: Towards Microbial Control ver 3.0
• [Presentation] 磁石を操る (2017)
  • Author(s): 田岡 東、福森義宏
  • Organizer: 第7回日本微生物学連盟フォーラム「微生物変わり者たちの素顔」
  • Invited
• [Presentation] Imaging of membrane vesicles for bacterial communication (2017)
  • Author(s): Taoka Azuma, Yousuke Kikuchi, Nozomu Obana, Masanori Toyohuku, Nobuhiko Nomura, Noriyuki Kodera, Toshio Ando, Yoshihiro Fukumori
  • Organizer: CHOZEN Project International Symposium on Atomic Force Microscopy at Solid-Liquid Interfaces,
  • Int'l Joint Research / Invited
• [Presentation] Actin-like MamK cytoskeleton tethers bacterial magnetosome organelle in a chain (2017)
  • Author(s): Azuma Taoka, Ayako Kiyokawa, Yousuke Kikuchi, Yoshihiro Fukumori
  • Organizer: International Symposium on "Harmonized supramolecular motility machinery and its diversity
  • Int'l Joint Research
• [Presentation] 全反射蛍光顕微鏡を用いたアクチン様蛋白質MamKの重合特性解析 (2017)
  • Author(s): 稲川 麻織奈、菊池 洋輔、田岡 東、福森 義宏
  • Organizer: 2017年度生命科学系学会合同年次大会（ConBio2017）
• [Presentation] 磁性細菌の生細胞内pHの蛍光イメージング (2017)
  • Author(s): 江口 友佳子、田岡 東、福森 義宏
  • Organizer: 2017年度生命科学系学会合同年次大会（ConBio2017）
• [Book] Biological Magnetic Materials and Applications (2018)
  • Author(s): Taoka, A. and Fukumori, Y.
  • Publisher: Springer
  • ISBN: 9789811080685
• [Remarks] 金沢大学生体分子生理学研究室ホームページ
  • URL: http://pronet.w3.kanazawa-u.ac.jp
• [Funded Workshop] The 6th International Meeting on Magnetotactic Bacteria (MTB 2018 meeting) (2018)