Organization and function of non-centrosomal microtubules array in polarized epithelial cells

• Project/Area Number: 25440094
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cell biology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Toya Mika 国立研究開発法人理化学研究所, 多細胞システム形成研究センター, 研究員 (80455360)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 極性上皮細胞 / 微小管編成 / 微小管マイナス端 / CAMSAP3/Nezha / 変異マウス / 小腸 / Caco-2細胞 / 立体培養系 / ノックアウトマウス / 三次元培養系 / Nezha/CAMSAP3

Outline of Final Research Achievements

Polarized epithelial cells organize characteristic array of non-centrosomal microtubules that align along apical to basal axis of the cells. The minus-ends of these microtubules face apically, and the plus-ends face towards the basal side. However, how this orientation is established remains elusive. CAMSAP3 is a member of CAMSAP (Calmodulin-regulated-spectrin-associated protein) family proteins that are known as microtubule minus-end binding proteins at non-centrosomal site. In this study, using CAMSAP3 mutant mice and 3D-cultured Caco-2 cells, we showed that CAMSAP3 localizes to the sub-apical region and tethers the minus-ends of microtubules to this site. In the absence of CAMSAP3 function, the orientation of cytoplasmic microtubules or positioning of cell organelle is perturbed. Obtained results demonstrate that apically localized CAMSAP3 determines the proper orientation of microtubules, and in turn that of cell organelles, in mammalian intestinal epithelial cells.

Research Products

• [Journal Article] CAMSAP3 orients the apical-to-basal polarity of microtubule arrays in epithelial cells (2016)
  • Author(s): Toya, M., Kobayashi, S., Kawasaki, M., Shioi, G., Kaneko, M., Ishiuchi, T., Misaki, K., Meng, W. and Takeichi, M.
  • Journal Title: Proc Natl Acad Sci USA Volume: 113 Issue: 2 Pages: 332-337
  • DOI: 10.1073/pnas.1520638113
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] CAMSAP3 orients the apical-to-basal polarity of microtubule arrays in epithelial cells (2016)
  • Author(s): Mika Toya and Masatoshi Takeichi
  • Organizer: Microtubules: From Atoms to Complex Systems
  • Place of Presentation: Heidelberg, Germany
  • Year and Date: 2016-05-29
  • Int'l Joint Research
• [Presentation] CAMSAP3 determines the apical-to-basal orientation of microtubules in polarized epithelial cells (2015)
  • Author(s): Mika Toya, Wenxiang Meng and Masatoshi Takeichi
  • Organizer: 第38会日本分子生物学会年会
  • Place of Presentation: 神戸ポートピアホテル 神戸市 兵庫県
  • Year and Date: 2015-12-03
  • Invited
• [Presentation] Nezha/CAMSAP3は極性上皮細胞における微小管の頂底軸配向に必須の役割を果たす (2015)
  • Author(s): 戸谷美夏, 孟文翔, 竹市雅俊
  • Organizer: 第67会日本細胞生物学会大会
  • Place of Presentation: タワーホール船堀 江戸川区 東京
  • Year and Date: 2015-06-30
• [Presentation] Nezha/CAMSAP3は極性上皮細胞における微小管の頂底軸配向に必須の役割を果たす (2014)
  • Author(s): 戸谷 美夏
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-27
• [Remarks] 上皮細胞の微小管配向の謎を解明 ー微小管結合タンパク質CAMSAP3が重要な働きー
  • URL: http://www.riken.jp/pr/press/2016/20160119_1/
• [Remarks] 上皮細胞の微小管を正しく配向させるメカニズム
  • URL: http://www.cdb.riken.jp/news/2016/researches/0119_9499.html
• [Remarks] ASCB Cell News
  • URL: http://www.ascb.org/cell-news-microtubules-organize-top-to-bottom-in-epithelial-cells-due-to-camsap3/