Phosphorylation mechanisms underlying the presynaptic active zone structure and function

• Project/Area Number: 18680032
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: University of Toyama
• Principal Investigator: OHTSUKA Toshihisa University of Toyama, 大学院医学薬学研究部, 准教授 (40401806)
• Project Period (FY): 2006 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥30,290,000 (Direct Cost: ¥23,300,000、Indirect Cost: ¥6,990,000); Fiscal Year 2008: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000); Fiscal Year 2007: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000); Fiscal Year 2006: ¥15,600,000 (Direct Cost: ¥12,000,000、Indirect Cost: ¥3,600,000)
• Keywords: シナプス / アクティブゾーン / CAST / ELKS / RIM1 / 神経伝達物質放出 / RIM / SAD / シナプス小胞 / リン酸化 / Bassoon

Research Abstract

神経終末アクティブゾーン(Active zone: AZ)に局在するSADキナーゼがAZたんぱく質であるRIM1のみならず、CASTをもリン酸化することを見出した。本リン酸化部位特異抗体による解析から、内在性のCASTもリン酸化されており、リン酸化されたCASTもAZに強く結合していることが明らかとなった。また、ラット上顎神経節細胞にリン酸化変異体を導入し、電気生理学的解析を行ったところ、

Research Products

• [Journal Article] SCRAPPER-Dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release (2007)
  • Author(s): Yao, I., Takagi, H., Ageta, H, Kahyo, T, Sato, S., Hatanaka, K., Fukuda, Y., Chiba, T., Morone, N., Yuasa, S., Inokuchi, K., Ohtsuka, T., MacGregor, G.R., Tanaka, K., Setou, M.
  • Journal Title: Cell 130 Pages: 943-957
• [Journal Article] Interaction between Munc13-1 and RIM is Critical for Glucagon-like Peptide-1-mediated Rescue of Exocytotic Defects in Munc13-1-deficient pancreatic {beta}-cells (2007)
  • Author(s): Kwan, E., Xie, L.Sheu, L.., Ohtsuka, T., and Gaisano, H.Y.
  • Journal Title: Diabetes 56 Pages: 2579-2588
• [Journal Article] Rab6 regulates transport and targeting of exocytotic carriers (2007)
  • Author(s): Grigoriev, I., Splinter, D., Keijzer, N., Wulf, P.,Demmers, J., Ohtsuka, T., Modesti, M., Maly, I., Grosveld, F., Hoogenraad, C. C., and Akhmanova, A.
  • Journal Title: Dev. Cell 13 Pages: 305-314
• [Journal Article] Three-dimensional architecture of presynaptic terminal cytomatrix (2007)
  • Author(s): Siksou, L.., Rostaing, P., Lechaire, J-P., Boudier, T., Ohtsuka, T., Fejtov , A., Kao, H-T., Greengard, P., Gundelfinger, E.D., Triller, A., and Marty, S.
  • Journal Title: J. Neurosci. 27 Pages: 6868-6877
• [Journal Article] The Active Zone Protein CAST Directly Associates with Ligand-of-Numb Protein X (2007)
  • Author(s): Higa Onaga, S., Tokoro, T., Inoue, E., Kitajima, I., and Ohtsuka, T.
  • Journal Title: Biochem. Biophys. Res. Commun. 354 Pages: 686-692
• [Journal Article] Localization of the Active Zone Proteins CAST, ELKS, and Piccolo at Neuromuscular Junctions (2007)
  • Author(s): Tokoro, T., Higa Onaga, S., Deguchi-Tawarada, M., Inoue, E., Kitajima, I., and Ohtsuka, T.
  • Journal Title: NeuroReport 18 Pages: 313-316
• [Journal Article] SCRAPPER-Dependent ubiquitination of active zone protein RIM1 regulates synaptic vesicle release. (2007)
  • Author(s): Yao, I.
  • Journal Title: Cell 130 Pages: 943-957
  • Peer Reviewed
• [Journal Article] Interaction between Munc13-1 and RIM is Critical for Glucagon-like Peptide-1-mediated Rescue of Exocytotic Defects in Munc13-1-deficient pancreatic {beta}-cells. (2007)
  • Author(s): Kwan, E.
  • Journal Title: Diabetes 56 Pages: 2579-2588
  • Peer Reviewed
• [Journal Article] Rab6A regulates transport and targeting of exocytotic carriers. (2007)
  • Author(s): Grigoriev, I.
  • Journal Title: Dev. Cell 13 Pages: 305-314
  • Peer Reviewed
• [Journal Article] Three-dimensional architecture of presynaptic terminal cytomatrix (2007)
  • Author(s): La, S., Rostaing
  • Journal Title: J. Neurosci. 27 Pages: 6868-6877
  • Peer Reviewed
• [Journal Article] The Active Zone Protein CAST Directly Associates with Ligand-of-Numb Protein X. (2007)
  • Author(s): Higa Onaga, S.
  • Journal Title: Biochem. Biophys. Res. Commun. 354 Pages: 686-692
  • Peer Reviewed
• [Journal Article] Localization of the Active Zone Proteins CAST, ELKS, and Piccolo at Neuromuscular Junctions. (2007)
  • Author(s): Tokoro, T
  • Journal Title: NeuroReport
• [Journal Article] The Active Zone Protein CAST Directly Associates with Ligand-of-Numb Protein X. (2007)
  • Author(s): Higa Onaga
  • Journal Title: Biochem. Biophys. Res. Gommun. 16;354 (3) Pages: 686-692
• [Journal Article] CLASPs attach microtube plus ends to the cell cortex through a complex with LL5β (2006)
  • Author(s): Lansgergen, G., Grigoriev, I., Minori-Kiyosue, Y., Ohtsuka, T., Higa, S., Kitajima, I., Demmers, J., Galjart, N., Houtsmuller, A.B., Grosveld, F. and Akhmanova, A.
  • Journal Title: Dev. Cell 11 Pages: 21-32
• [Journal Article] SAD: a presynaptic kinase associated with synaptic vesicles and the active zone cytomatrix that regulates neurotransmitter release (2006)
  • Author(s): Inoue, E., Mochida, S., Takagi, H., Higa, S., Deguchi-Tawarada, M., Takao-Rikitsu, E., Inoue, M., Yao, I., Takeuchi, K., Kitajima, I., Setou, M., Ohtsuka, T., and Takai, Y.
  • Journal Title: Neuron 50 Pages: 261-275
• [Journal Article] ELKS, a protein structurally related to the active zone protein CAST, regulates Ca2+-dependent exocytosis from PC12 cells (2006)
  • Author(s): Inoue, E, Deguchi-Tawarada, M., Takao-Rikitsu, E.., Inoue, M., Kitajima, I., Ohtsuka, T., and Takai, Y.
  • Journal Title: Genes Cells 11 Pages: 659-672
• [Journal Article] Active Zone Protein CAST is a Component of Conventional and Ribbon Synapses in Mouse Retina (2006)
  • Author(s): Deguchi-Tawarada, M., Inoue, E., Takao-Rikitsu, E., Inoue, M., Kitajima,I., Ohtsuka, T., and Takai, Y.
  • Journal Title: J. Comp. Neurol. 495 Pages: 480-496
• [Journal Article] Active Zone Protein CAST is a Component of Conventional and Ribbon Synapses in Mouse Retina. (2006)
  • Author(s): Deguchi-Tawarada, M
  • Journal Title: J. Comp. Neurol. 495 (4) Pages: 480-496
• [Journal Article] ELKS, a protein structurally related to the active zone protein CAST, regulates Ca2+-dependent exocytosis from PC12 cells. (2006)
  • Author(s): Inoue, E
  • Journal Title: Genes Cells 11 (6) Pages: 659-672
• [Journal Article] SAD : a presynaptic kinase associated with synaptic vesicles and the active zone cytomatrix that regulates neurotransmitter release. (2006)
  • Author(s): Inoue. E
  • Journal Title: Neuron 50 (2) Pages: 261-275
• [Journal Article] CLASPs attach microtube plus ends to the cell cortex through a complex with LL5β. (2006)
  • Author(s): Lansbergen, G
  • Journal Title: Dev. Cell 11 (1) Pages: 21-32
• [Presentation] 神経終末アクティブゾーンにおけるシグナル伝達 (2008)
  • Author(s): 大塚稔久
  • Organizer: 第31回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2008-07-11
• [Presentation] 神経終末アクティブゾーンの分子解剖学 (2008)
  • Author(s): 大塚稔久
  • Organizer: 特定領域研究 公開シンポジウム 「生体膜トランスポートソームの分子構築と生理機能」
  • Place of Presentation: 東京
  • Year and Date: 2008-03-22
• [Presentation] 神経シナプスアティブ・ゾーンの生化学 (2007)
  • Author(s): 大塚稔久
  • Organizer: 第30回日本分子生物会年会
  • Place of Presentation: 横浜
• [Presentation] SADキナーゼのマウス神経筋接合部における局在 (2007)
  • Author(s): 所崇 吉田淑子 比嘉進 大塚稔久
  • Organizer: Neuro2007
  • Place of Presentation: 横浜
• [Presentation] SADキナーゼによるアクチブゾーン特異蛋白質BassoonおよびPiccoloのリン酸化 (2007)
  • Author(s): Susumu Higa, 所崇, 北島勲, 大塚稔久
  • Organizer: Neuro2007
  • Place of Presentation: 横浜
• [Presentation] Biochemistry of the presynaptic active zone (regular talk). (2007)
  • Author(s): Toshihisa Ohtsuka
  • Organizer: Gordon Research Conferences
  • Place of Presentation: NH, USA.
• [Presentation] マウス神経筋接合部におけるアクティブゾーン蛋白質CAST/ELKSの局在 (2006)
  • Author(s): 所崇 比嘉進 北島勲 大塚稔久
  • Organizer: 分子生物学の未来 日本分子生物学会2006 フォーラム
  • Place of Presentation: 名古屋
• [Presentation] 学会 The active zone protein CAST directly associates with the Ligand of Numb X (2006)
  • Author(s): 比嘉進 井上英二 所崇 北島勲 大塚稔久
  • Organizer: 分子生物学の未来 日本分子生物学会2006 フォーラム
  • Place of Presentation: 名古屋