Elucidating the mechanism of the Wnt/PCP signaling pathway by the genome editing technology

• Project/Area Number: 15K07054
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cell biology
• Research Institution: Kumamoto University
• Principal Investigator: Kikuchi Koji 熊本大学, 大学院生命科学研究部(医), 講師 (70457290)
• Co-Investigator(Renkei-kenkyūsha): TANAKA Tsubasa 熊本大学, 発生医学研究所, 助教 (00392027)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: Wnt5aシグナル経路 / 微小管 / 微小管結合タンパク質 / Dishevelled / キネシン / 細胞接着 / 細胞運動 / 平面内細胞極性 / Wnt/PCPシグナル経路 / 微小管リモデリング / 微小管結合蛋白質 / 前後極性 / 上皮組織 / 微小管ダイナミクス / ゲノム編集 / Wnt/平面内細胞極性経路 / Dvl

Outline of Final Research Achievements

Wnt5a signaling through a β-catenin-independent pathway promotes microtubule (MT) remodeling during cell-substrate adhesion, cell migration, and planer cell polarity (PCP) formation, all of which are essential features of tissue morphogenesis. Although Wnt5a signaling and MT remodeling are known to form an interdependent regulatory loop, the underlying mechanism remains unknown. Here we show that Map7/7D1 cooperate with Kif5b to coordinate a feedback loop between Dvl dynamics and MT remodeling in the Wnt5a signaling pathway, and that the role of Map7/7D1 family proteins in Dvl/Dsh localization is evolutionarily conserved.
As a methodological highlight, we adapted a CRISPR-Cas9-mediated genome editing technology to generate knock-in HeLa cells and fly strains. As the overexpression of Map7/7D1 family proteins is reported to induce aberrant MT bundling, this strategy allowed us to reliably analyze behaviors of endogenous Map7/7D1 family proteins in cells and tissues.

Research Products

• [Journal Article] Map7/7D1 and Dvl form a feedback loop that facilitates microtubule remodeling and Wnt5a signaling. (2018)
  • Author(s): Kikuchi, K., Nakamura, A., Arata, M., Shi, D., Nakagawa, M., Tanaka, T., Uemura, T., Fujimori, T., Kikuchi, A., Uezu, A., Sakamoto, Y., and Nakanishi, H.
  • Journal Title: EMBO Rep. Volume: 印刷中
  • Peer Reviewed
• [Journal Article] Effects of various spacers between biotin and the phospholipid headgroup on immobilization and sedimentation of biotinylated phospholipid-containing liposomes facilitated by avidin-biotin interactions. (2017)
  • Author(s): Sakamoto, Y., Kikuchi, K., Umeda, K. & Nakanishi, H.
  • Journal Title: Journal of biochemistry Volume: 未定 Issue: 3 Pages: 221-226
  • DOI: 10.1093/jb/mvx016
  • NAID: 40021326130
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Map7/7D1 and Dvl form a feedback loop that facilitates microtubule remodeling and Wnt5a signaling. (2018)
  • Author(s): Kikuchi, K., Nakamura, A., Arata, M., Shi, D., Nakagawa, M., Tanaka, T., Uemura, T., Fujimori. T., Kikuchi, A., and Nakanishi, H.
  • Organizer: KEY FORUM: The 3rd International Symposium on Stem Cell Traits and Developmental Systems.
• [Presentation] Identification of Novel Microtubule-associated Proteins that Contribute to the Regulation of the Wnt/PCP Signaling Pathway. (2017)
  • Author(s): Koji Kikuchi, Tsubasa Tanaka, Masaki Arata, Dongbo Shi, Akira Nakamura, Tadashi Uemura, Toshihiko Fujimori, Hiroyuki Nakanishi
  • Organizer: The 1st ABiS Symposium Towards the Future of Advanced Bioimaging for Life Sciences
  • Place of Presentation: 自然科学研究機構・岡崎コンファレンスセンター
  • Year and Date: 2017-02-19
• [Presentation] Coordination of cytoskeletal remodeling and Dishevelled localization in Wnt/PCP signaling by conserved microtubule-associated proteins. (2017)
  • Author(s): Kikuchi, K., Nakamura, A., Arata, M., Shi, D., Nakagawa, M., Tanaka, T., Uemura, T., Fujimori. T., Kikuchi, A., and Nakanishi, H.
  • Organizer: ConBio2017
• [Presentation] Novel regulation mechanisms of microtubule dynamics during neurite outgrowth. (2017)
  • Author(s): Kikuchi, K., Kuramoto, T., Uezu, A., Saito, T., Hisanaga, S., and Nakanishi, H.
  • Organizer: 第69回日本細胞生物学会大会
• [Presentation] Identification of novel microtubule-associated proteins that contribute to the regulation of the Wnt/PCP signaling pathway. (2016)
  • Author(s): 菊池 浩二、田中 翼、新田 昌輝、石 東博、中村 輝、上村 匡、藤森 俊彦、中西 宏之
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2016-11-30
• [Presentation] Identification of novel microtubuleassociated proteins that contribute to the epithelial morphogenesis through the Wnt/PCP signaling pathway. (2016)
  • Author(s): Koji Kikuchi, Tsubasa Tanaka, Masaki Arata, Dongbo Shi, Akira Nakamura, Tadashi Uemura, Toshihiko Fujimori, Hiroyuki Nakanishi
  • Organizer: JDRC12
  • Place of Presentation: 立教大学
  • Year and Date: 2016-09-09
• [Presentation] 微小管結合蛋白質によるWnt/平面内細胞極性経路を介した上皮組織の構築メカニズム (2016)
  • Author(s): 菊池 浩二、田中 翼、新田 昌輝、石 東博、中村 輝、上村 匡、藤森 俊彦、中西 宏之
  • Organizer: 第68回日本細胞生物学会大会
  • Place of Presentation: 京都テルサ
  • Year and Date: 2016-06-15
• [Presentation] 神経突起伸長を制御する新規微小管結合タンパク質の同定 (2015)
  • Author(s): 菊池 浩二
  • Organizer: BMB2015
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-01
• [Presentation] 微小管結合蛋白質によるWnt/平面内細胞極性経路を介した細胞骨格のダイナミクス制御機構 (2015)
  • Author(s): 菊池 浩二
  • Organizer: 第67回日本細胞生物学会大会
  • Place of Presentation: タワーホール船堀
  • Year and Date: 2015-06-30
• [Remarks] 研究室ホームページ
  • URL: https://sites.google.com/site/lab1yakurihp/
• [Remarks] Researchmap
  • URL: http://researchmap.jp/read0147459/
• [Remarks] Researchgate
  • URL: https://www.researchgate.net/profile/Koji_Kikuchi/
• [Remarks] ORCID
  • URL: http://orcid.org/0000-0002-7616-8200
• [Remarks] ResearcherID
  • URL: http://www.researcherid.com/rid/W-1048-2017
• [Remarks] Researchmapの個人ページ
  • URL: http://researchmap.jp/read0147459/
• [Remarks] Researchgateの個人ページ
  • URL: https://www.researchgate.net/profile/Koji_Kikuchi/
• [Remarks] 所属研究室のホームページ
  • URL: https://sites.google.com/site/lab1yakurihp/
• [Remarks] 所属研究機関が作成した研究室紹介のページ
  • URL: http://www.medphas.kumamoto-u.ac.jp/research/bunya/8.html
• [Remarks] researchmapの個人ページ
  • URL: http://researchmap.jp/read0147459/