Regulation of primary ciliogenesis by Hippo signalling

• Project/Area Number: 25840065
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Cell biology
• Research Institution: Osaka University
• Principal Investigator: CHIBA Shuhei 大阪大学, 医学(系)研究科(研究院), 特任助教(常勤) (60572493)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 一次繊毛 / 細胞周期 / NDR / Rabin8 / Hippo / 基底小体 / 繊毛症 / Rab8 / Distal Appendages / 増殖と分化 / 膜輸送 / Hippo経路 / 中心体サイクル / 繊毛鞘

Outline of Final Research Achievements

NDR2, an effector molecule for mammalian Hippo signalling, was identified as the causal gene for a canine ciliopathy, early retinal degeneration. Little is known how NDR2 regulates cilium formation. Ciliary membranes are generated by transport and fusion of Golgi-derived vesicles to the pericentrosome, a process requiring Rab11-mediated recruitment of Rabin8, subsequent activation of Rab8 and Rabin8 binding to Sec15. I found that NDR2 phosphorylates Rabin8 and defects in this phosphorylation impair preciliary membrane assembly and ciliogenesis. NDR2-mediated Rabin8 phosphorylation is crucial for ciliogenesis by triggering the switch in binding specificity of Rabin8 from PS to Sec15, thereby promoting local activation of Rab8 and ciliary membrane formation. Additionally, I identified a certain member of lipid kinase as a new physiological substrate for NDR2. These results suggest that the NDR2 is crucial for ciliogenesis by regulating intracellular vesicle trafficking.

Research Products

• [Journal Article] Binding to Cep164, b ut not EB1, is essential for centriolar localization of TTBK2 and its function in ciliogenesis. Genes Cells. (2014)
  • Author(s): Oda, T., Chiba, S., Nagai, T., and Mizuno, K.
  • Journal Title: Genes Cells. Volume: 19 Issue: 12 Pages: 927-940
  • DOI: 10.1111/gtc.12191
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] NDR2-mediated Rabin8 phosphorylation is crucial for ciliogenesis by switching binding specificity from phosphatidylserine to Secl5. (2013)
  • Author(s): Chiba, S., Amagai, Y., Homma, Y., Fukuda, M., and Mizuno, K.
  • Journal Title: EMBO J. Volume: 32 Issue: 6 Pages: 874-885
  • DOI: 10.1038/emboj.2013.32
  • Peer Reviewed
• [Journal Article] Furry promotes acetylation of microtubules in the mitotic spindle via inhibition of SIRT2 tubulin-deacetylase (2013)
  • Author(s): Tomoaki Nagai, Masanori Ikeda, Shuhei Chiba, Shin-ichiro Kanno and Kensaku Mizuno
  • Journal Title: J Cell Sci Volume: 126 Issue: 19 Pages: 4369-80
  • DOI: 10.1242/jcs.127209
  • Peer Reviewed
• [Journal Article] Genetically encoded calcium indicator illuminates calcium dynamics in primary cilia. (2013)
  • Author(s): Su S, Phua SC, DeRose R, Chiba S, Narita K, Kalugin PN, Katada T, Kontani K, Takeda S, Inoue T.
  • Journal Title: Nat. Methods Volume: 10 Issue: 11 Pages: 1105-1107
  • DOI: 10.1038/nmeth.2647
  • Peer Reviewed
• [Presentation] 細胞増殖抑制時の一次繊毛形成における TTBK2 キナーゼの機能解析 (2015)
  • Author(s): 永井友朗、小田聡明、千葉秀平、水野健作
  • Organizer: 日本生化学会東北支部 第81回例会・シンポジウム
  • Place of Presentation: 東北大学片平桜さくらホール (仙台)
  • Year and Date: 2015-05-09
• [Presentation] Cep164, but not EB1, is essential for centriolar localization of TTBK2 and its function in ciliogenesis (2014)
  • Author(s): Nagai, T., Oda, T., Chiba, S., Mizuno, K.
  • Organizer: 2014 ASCB/IFCB meeting
  • Place of Presentation: Pennsylvania Convention Center (Philadelphia, USA)
  • Year and Date: 2014-12-06 – 2014-12-10
• [Presentation] TTBK2はCep164と結合することで中心体に局在し、一次繊毛形成を促進する (2014)
  • Author(s): 小田 聡明, 千葉 秀平, 永井 友朗, 水野 健作
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜 (横浜)
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 細胞周期依存的な中心体-基小体変換機構 (2014)
  • Author(s): 水野健作、小田聡明、永井友朗、千葉秀平
  • Organizer: 新学術研究領域「シリア・中心体による生体情報フローの制御」 第3回領域会議
  • Place of Presentation: 倉敷シーサイドホテル (倉敷)
  • Year and Date: 2014-07-17 – 2014-07-18
• [Presentation] 一次繊毛形成におけるEB1の機能 (2014)
  • Author(s): 小田聡明、千葉秀平、永井友朗、水野健作
  • Organizer: 第5回繊毛研究会
  • Place of Presentation: 浜松市研究交流センター (浜松)
  • Year and Date: 2014-05-24 – 2014-05-25
• [Presentation] 一次繊毛形成を担うRabin8のリン酸化制御解析 (2013)
  • Author(s): 千葉秀平、天貝佑太、本間裕太、福田光則、水野健作
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: 横浜
  • Invited
• [Presentation] NDR2-mediated Rabin8 phosphorylation is crucial for ciliogenesis by switching binding specificity from phosphatidylserine to Sec15. (2013)
  • Author(s): Chiba, S., Amagai, Y., Homma, Y., Fukuda, M., Mizuno, K.
  • Organizer: FASEB Science Research Conferences “ Biology of Cilia & Flagella ”
  • Place of Presentation: 米国、ニューヨーク州
• [Presentation] NDR2 によるRabin8 のリン酸化と一次繊毛形成における機能 (2013)
  • Author(s): 千葉秀平、天貝佑太、本間悠太、福田光則、水野健作
  • Organizer: 日本生化学会 東北支部 第79回例会
  • Place of Presentation: 仙台
• [Book] 実験医学 (2014)
  • Author(s): 千葉秀平、井上尊生
  • Total Pages: 6
  • Publisher: 羊土社
• [Book] 実験医学 2014年5月号 Vol.32 No.8 (2014)
  • Author(s): 千葉秀平、井上尊生
  • Total Pages: 6
  • Publisher: 羊土社
• [Book] 実験医学増刊「細胞周期による高次生命現象の制御と疾患」Vol.31 No.2 (2013)
  • Author(s): 千葉秀平、天貝佑太、水野健作
  • Total Pages: 5
  • Publisher: 羊土社
• [Remarks] 東北大学 大学院生命科学研究科 情報伝達分子解析分野 水野研究室
  • URL: http://www.biology.tohoku.ac.jp/lab-www/mizuno_lab/
• [Remarks] 東北大学生命科学研究科
  • URL: http://www.lifesci.tohoku.ac.jp/research_ja/15442/