Mechanical manipulation and in vitro reconstitution of microtubule severing

• Project/Area Number: 18KK0195
• Research Category: Fund for the Promotion of Joint International Research (Fostering Joint International Research (B))
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
• Research Institution: Nagoya University
• Principal Investigator: Nakamura Masayoshi 名古屋大学, トランスフォーマティブ生命分子研究所, 特任准教授 (40553409)
• Co-Investigator(Kenkyū-buntansha): 吉成 晃 名古屋大学, トランスフォーマティブ生命分子研究所, 学振特別研究員(PD) (00829872) ; 橋本 隆 奈良先端科学技術大学院大学, 先端科学技術研究科, 教授 (80180826)
• Project Period (FY): 2018-10-09 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000); Fiscal Year 2021: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2020: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2019: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2018: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 微小管 / 細胞骨格 / カタニン / チューブリン / シロイヌナズナ / 植物

Outline of Final Research Achievements

Microtubule severing through katanin activities plays a crucial role in the organization of cortical microtubules in plant cells in response to developmental and environmental cues. Analyses of microtubule dynamics have elucidated the sites of microtubule cleavage and the duration required for cleavage in interphase epidermal cells of plants. However, the molecular mechanisms underlying katanin localization and severing activity remain unresolved. In this study, alongside conventional genetic and biochemical analyses, we developed a live cell imaging method and an in vitro reconstruction system, demonstrating that the p80 regulatory subunit regulates the microtubule severing site by p60.

Academic Significance and Societal Importance of the Research Achievements

カタニンによる微小管切断は、発生過程や環境に応答しながら、動的な表層微小管の配向パターン形成において重要な役割を果たしている。微小管動態解析により植物体表面の間期表皮細胞における微小管切断は微小管形成部位と交差部位で引き起こされることが知られている。しかしながら、生命活動に重要なカタニンの局在性や切断活性制御の分子基盤は明らかとなっていなかった。本研究では、カタニンによる微小管切断が起こる場所に着目した国際共同研究を展開し、切断箇所の決定メカニズムを明らかにした。

Research Products

• [Int'l Joint Research] Carnegie Institution for Science(米国)
• [Int'l Joint Research] Carnegie Institution for Science(米国)
• [Int'l Joint Research] Institute of Biotechnology AS CR(チェコ)
• [Int'l Joint Research] Carnegie Institution for Science(米国)
• [Int'l Joint Research] Institute of Biotechnology AS CR(チェコ)
• [Journal Article] Near‐infrared imaging of phytochrome‐derived autofluorescence in plant nuclei (2024)
  • Author(s): Yoshinari Akira、Isoda Reika、Yagi Noriyoshi、Sato Yoshikatsu、Lindeboom Jelmer J.、Ehrhardt David W.、Frommer Wolf B.、Nakamura Masayoshi
  • Journal Title: The Plant Journal Volume: - Issue: 5 Pages: 1699-1712
  • DOI: 10.1111/tpj.16699
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Finding a right place to cut: How katanin is targeted to cellular severing sites (2022)
  • Author(s): Masayoshi Nakamura, Noriyoshi Yagi, Takashi Hashimoto
  • Journal Title: Quantitative Plant Biology Volume: 3
  • DOI: 10.1017/qpb.2022.2
  • Peer Reviewed / Open Access
• [Journal Article] An anchoring complex recruits katanin for microtubule severing at the plant cortical nucleation sites (2021)
  • Author(s): Noriyoshi Yagi , Takehide Kato , Sachihiro Matsunaga , David W. Ehrhardt , Masayoshi Nakamura , Takashi Hashimoto
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 3684-3684
  • DOI: 10.1038/s41467-021-24067-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Using Genetically Encoded Fluorescent Biosensors for Quantitative In Vivo Imaging (2020)
  • Author(s): Yoshinari Akira、Moe-Lange Jacob、Kleist Thomas J.、Cartwright Heather N.、Quint David A.、Ehrhardt David W.、Frommer Wolf B.、Nakamura Masayoshi
  • Journal Title: Arabidopsis Protocols, Methods in Molecular Biology Volume: 2200 Pages: 303-322
  • DOI: 10.1007/978-1-0716-0880-7_14
  • ISBN: 9781071608791, 9781071608807
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Covalent Self-labeling of Tagged Proteins with Chemical Fluorescent Dyes in BY-2 Cells and Arabidopsis Seedlings (2020)
  • Author(s): Iwatate, J.R., Yoshinari, A., Yagi, N., Grzybowski, M., Ogasawara, H., Kamiya, M., Komatsu, T., Taki, M., Yamaguchi, S., Frommer, W.B. and Nakamura, M.
  • Journal Title: The Plant Cell Volume: 32 Issue: 10 Pages: 3081-3094
  • DOI: 10.1105/tpc.20.00439
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mechanistic Insights into Plant Chiral Growth (2020)
  • Author(s): Nakamura, M. and Hashimoto, T.
  • Journal Title: Symmetry Volume: 12 Issue: 12 Pages: 2056-2056
  • DOI: 10.3390/sym12122056
  • Peer Reviewed
• [Journal Article] CLASP stabilization of plus ends created by severing promotes microtubule creation and reorientation (2019)
  • Author(s): Lindeboom Jelmer J.、Nakamura Masayoshi、Saltini Marco、Hibbel Anneke、Walia Ankit、Ketelaar Tijs、Emons Anne Mie C.、Sedbrook John C.、Kirik Viktor、Mulder Bela M.、Ehrhardt David W.
  • Journal Title: The Journal of Cell Biology Volume: 218 Issue: 1 Pages: 190-205
  • DOI: 10.1083/jcb.201805047
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Elucidating molecular mechanisms of microtubule nucleation, severing, and regulation of microtubule dynamics during the organization of cortical microtubule arrays in plants (2023)
  • Author(s): Masayoshi Nakamura
  • Organizer: The 49th NAITO Conference on Frontiers of Microtubule and Its-related Motors
  • Int'l Joint Research / Invited
• [Presentation] An anchoring complex stabilizes the association of daughter microtubule minus ends to their nucleation sites (2022)
  • Author(s): Noriyoshi Yagi, Masayoshi Nakamura
  • Organizer: Gordon Research Conference, Plant and Microbial Cytoskeleton
  • Int'l Joint Research
• [Remarks] The anchor and the sword:
  • URL: https://www.itbm.nagoya-u.ac.jp/en_backup/research/2021/06/post-35.php