Elucidation of phosphorylation signal regulating the cell polarity

• Project/Area Number: 25251021
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Cell biology
• Research Institution: Nagoya University
• Principal Investigator: Kaibuchi Kozo 名古屋大学, 医学系研究科, 教授 (00169377)
• Co-Investigator(Kenkyū-buntansha): 西岡 朋生 名古屋大学, 医学(系)研究科(研究院), 助教 (70435105) ; 天野 睦紀 名古屋大学, 医学(系)研究科(研究院), 准教授 (90304170) ; 黒田 啓介 名古屋大学, 医学(系)研究科(研究院), 特任助教 (80631431) ; 中牟田 信一 名古屋大学, 医学(系)研究科(研究院), 助教 (20647474) ; 渡辺 崇 名古屋大学, 医学(系)研究科(研究院), 講師 (10402562)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥46,930,000 (Direct Cost: ¥36,100,000、Indirect Cost: ¥10,830,000); Fiscal Year 2015: ¥13,520,000 (Direct Cost: ¥10,400,000、Indirect Cost: ¥3,120,000); Fiscal Year 2014: ¥13,520,000 (Direct Cost: ¥10,400,000、Indirect Cost: ¥3,120,000); Fiscal Year 2013: ¥19,890,000 (Direct Cost: ¥15,300,000、Indirect Cost: ¥4,590,000)
• Keywords: 細胞内情報伝達 / 細胞極性 / リン酸化 / 質量分析

Outline of Final Research Achievements

The establishment and maintenance of cell polarity is essential to a wide range of biological processes, including axon formation and cell migration. In elucidating of molecular basis of cell polarization, cell polarity proteins such as PAR complex, and Scrib complex have been studied. In these studies, various kinases have been shown to act as cell polarity regulators. However, how these kinases regulate cell polarization is poorly understood. In this study, we used our recently developed proteomic system and identified numerous candidate substrates downstream of polarity-related kinases. Based on the substrate screening results, we analyzed the spatiotemporal dynamics and function of the identified substrates during cell migration. Recently, we also revealed how neurons can grow only a single axon while preventing the formation of unnecessary axons during neuronal polarization.

Research Products

• [Journal Article] Discovery of Long-Range Inhibitory Signaling to Ensure Single Axon Formation (2017)
  • Author(s): Takano T, Wu M, Nakamuta S, Naoki H, Ishizawa N, Namba T, Watanabe T, Xu C, Hamaguchi T, Yura Y, Amano M, Hahn KM, Kaibuchi K
  • Journal Title: Nature Communications Volume: 印刷中
  • NAID: 120006325188
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Phosphorylation Signals in Striatal Medium Spiny Neurons (2016)
  • Author(s): Nagai T, Yoshimoto J, Kannon T, Kuroda K, Kaibuchi K
  • Journal Title: Trends in Pharmacological Sciences Volume: 37 Issue: 10 Pages: 858-71
  • DOI: 10.1016/j.tips.2016.07.003
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Focused Proteomics Revealed a Novel Rho-kinase Signaling Pathway in the Heart (2016)
  • Author(s): Yura Y, Amano M, Takefuji M, Bando T, Suzuki K, Kato K, Hamaguchi T, Hasanuzzaman Shohag M, Takano T, Funahashi Y, Nakamuta S, Kuroda K, Nishioka T, Murohara T, Kaibuchi K
  • Journal Title: Cell Structure and Function Volume: 41 Issue: 2 Pages: 105-120
  • DOI: 10.1247/csf.16011
  • NAID: 130005405432
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] PAR3-aPKC regulates Tiam1 by modulating suppressive internal interactions. (2016)
  • Author(s): Matsuzawa, K., Akita, H., Watanabe, T., Kakeno, M., Matsui, T., Wang, S., and Kaibuchi, K.
  • Journal Title: Mol Biol Cell Volume: in press Issue: 9 Pages: 1511-23
  • DOI: 10.1091/mbc.e15-09-0670
  • Peer Reviewed
• [Journal Article] Phosphoproteomics of the Dopamine Pathway Enables Discovery of Rap1 Activation as a Reward Signal In Vivo (2016)
  • Author(s): Nagai T, Nakamuta S, Kuroda K, Nakauchi S, Nishioka T, Takano T, Zhang X, Tsuboi D, Funahashi Y, Nakano T, Yoshimoto J, Kobayashi K, Uchigashima M, Watanabe M, Miura M, Nishi A, Kobayashi K, Yamada K, Amano M, Kaibuchi K
  • Journal Title: Neuron Volume: 89 (3) Issue: 3 Pages: 550-65
  • DOI: 10.1016/j.neuron.2015.12.019
  • Peer Reviewed / Open Access
• [Journal Article] Radial Glial Cell-Neuron Interaction Directs Axon Formation at the Opposite Side of the Neuron from the Contact Site. (2015)
  • Author(s): Xu, C.;Funahashi, Y.;Watanabe, T.;Takano, T.;Nakamuta, S.;Namba, T.;Kaibuchi, K.
  • Journal Title: J NEUROSCI Volume: 35 Issue: 43 Pages: 14517-14532
  • DOI: 10.1523/jneurosci.1266-15.2015
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] TTBK2 with EB1/3 regulates microtubule dynamics in migrating cells through KIF2A phosphorylation. (2015)
  • Author(s): Watanabe, T., Kakeno, M., Matsui, T., Sugiyama, I., Arimura, N., Matsuzawa, K., Shirahige, A., Ishidate, F., Nishioka, T., Taya, S., Hoshino, M., and Kaibuchi, K.
  • Journal Title: J Cell Biol Volume: 210 Issue: 5 Pages: 737-751
  • DOI: 10.1083/jcb.201412075
  • Peer Reviewed / Open Access
• [Journal Article] Extracellular and Intracellular Signaling for Neuronal Polarity. (2015)
  • Author(s): Namba, T.;Funahashi, Y.; Nakamuta, S.;Xu, C.;Takano, T.;Kaibuchi, K.
  • Journal Title: Physiol Rev Volume: 95 Issue: 3 Pages: 995-1024
  • DOI: 10.1152/physrev.00025.2014
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Phosphoproteomic Analysis Using the WW and FHA Domains as Biological Filters (2015)
  • Author(s): Hasanuzzaman Shohag, M., Nishioka, T., Uddin Ahammad, R., Nakamuta, S., Yura, Y., Hamaguchi, T., Kaibuchi, K., and Amano, M.
  • Journal Title: Cell Structure and Function Volume: 40 Issue: 2 Pages: 95-104
  • DOI: 10.1247/csf.15004
  • NAID: 130005094036
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed
• [Journal Article] Kinase-interacting substrate screening is a novel method to identify kinase substrates. (2015)
  • Author(s): Amano, M., Hamaguchi, T., Shohag, M. H., Kozawa, K., Kato, K., Zhang, X., Yura, Y., Matsuura, Y., Kataoka, C., Nishioka, T., and Kaibuchi, K.
  • Journal Title: J Cell Biol Volume: 209 Issue: 6 Pages: 895-912
  • DOI: 10.1083/jcb.201412008
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Neuronal polarization. (2015)
  • Author(s): Takano, T.; Xu, C.; Funahashi, Y.;Namba, T.;Kaibuchi, K.
  • Journal Title: Development Volume: 142 Issue: 12 Pages: 2088-2093
  • DOI: 10.1242/dev.114454
  • NAID: 40021797273
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] PAR3 and aPKC regulate Golgi organization through CLASP2 phosphorylation to generate cell polarity. (2015)
  • Author(s): Matsui, T., Watanabe, T., Matsuzawa, K., Kakeno, M., Okumura, N., Sugiyama, I., Itoh, N., and Kaibuchi, K.
  • Journal Title: Molecular Biology of the Cell Volume: 26 Issue: 4 Pages: 751-761
  • DOI: 10.1091/mbc.e14-09-1382
  • Peer Reviewed
• [Journal Article] <i>In vivo</i> Screening for Substrates of Protein Kinase A Using a Combination of Proteomic Approaches and Pharmacological Modulation of Kinase Activity (2015)
  • Author(s): Hamaguchi, T., Nakamuta, S., Funahashi, Y., Takano, T., Nishioka, T., Shohag, M.H., Yura, Y., Kaibuchi, K., and Amano, M.
  • Journal Title: Cell Structure and Function Volume: 40 Issue: 1 Pages: 1-12
  • DOI: 10.1247/csf.14014
  • NAID: 130004712922
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Developing novel methods to search for substrates of protein kinases such as Rho-kinase (2015)
  • Author(s): Nishioka T, Shohag MH, Amano M, Kaibuchi K
  • Journal Title: Biochimica et Biophysica Acta Volume: - Issue: 10 Pages: 1663-1666
  • DOI: 10.1016/j.bbapap.2015.03.001
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Disrupted-in-schizophrenia 1 regulates transport of ITPR1 mRNA for synaptic plasticity. (2015)
  • Author(s): Tsuboi D, Kuroda K, Tanaka M, Namba T, Iizuka Y, Taya S, Shinoda T, Hikita T, Muraoka S, Iizuka M, Nimura A, Mizoguchi A, Shiina N, Sokabe M, Okano H, Mikoshiba K, Kaibuchi K
  • Journal Title: Nature Neuroscience Volume: in press Issue: 5 Pages: 698-707
  • DOI: 10.1038/nn.3984
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Neuronal polarization in vivo: Growing in a complex environment. (2014)
  • Author(s): Funahashi, Y., Namba, T., Nakamuta, S., and Kaibuchi, K.
  • Journal Title: Current opinion in neurobiology Volume: 27 Pages: 215-223
  • DOI: 10.1016/j.conb.2014.04.009
  • Peer Reviewed / Open Access
• [Journal Article] Plk1 Phosphorylates CLIP-170 and Regulates Its Binding to Microtubules for Chromosome Alignment (2014)
  • Author(s): Kaneko M., Matsuzawa K., Matsui T., Akita H., Sugiyama I., Ishidate F., Nakano A., Takashima S., Goto H., Inagaki M., Kaibuchi K., and Watanabe T.
  • Journal Title: Cell Structure and Function Volume: 39 Issue: 1 Pages: 45-59
  • DOI: 10.1247/csf.14001
  • NAID: 130004053900
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed / Open Access
• [Journal Article] Pioneering Axons Regulate Neuronal Polarization in the Developing Cerebral Cortex. (2014)
  • Author(s): Namba T, Kibe Y, Funahashi Y, Nakamuta S, Takano T, Ueno T, Shimada A, Kozawa S, Okamoto M, Shimoda Y, Oda K, Wada Y, Masuda T, Sakakibara A, Igarashi M, Miyata T, F-S Catherine, Takeuchi K, Kaibuchi K.
  • Journal Title: Neuron Volume: 81(4) Issue: 4 Pages: 814-829
  • DOI: 10.1016/j.neuron.2013.12.015
  • Peer Reviewed / Open Access
• [Journal Article] ERK2-mediated phosphorylation of Par3 regulates neuronal polarization. (2013)
  • Author(s): Yasuhiro Funahashi, Takashi Namba, Shin Fujisue, Norimichi Itoh, Shinichi Nakamuta, Katsuhiro Kato, Akiko Shimada, Chundi Xu, Wei Shan, Tomoki Nishioka, Kozo Kaibuchi
  • Journal Title: The Journal of Neuroscience Volume: 33 Issue: 33 Pages: 13270-13285
  • DOI: 10.1523/jneurosci.4210-12.2013
  • Peer Reviewed
• [Presentation] KIOSS法によるPKCのリン酸化基質スクリーニングと その候補分子の解析 (2016)
  • Author(s): 西岡 朋生
  • Organizer: 第68回日本細胞生物学会・第11回日本ケミカルバイオロジー学会 合同大会
  • Place of Presentation: 京都テルサ（京都府京都市）
  • Year and Date: 2016-06-15
• [Presentation] Extracellular and intracellular signaling for neuronal polarity (2015)
  • Author(s): Kozo Kaibuchi
  • Organizer: EMBO Cortical Neural Development
  • Place of Presentation: Taipei (Taiwan)
  • Year and Date: 2015-12-04
  • Int'l Joint Research / Invited
• [Presentation] 生体内における神経細胞の極性形成機構 (2015)
  • Author(s): 貝淵 弘三
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会 合同年会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] Neuronal polarization in vivo: Growing in a complex environment (2015)
  • Author(s): Kozo Kaibuchi
  • Organizer: EMBO Workshop Cotrical Development in Health and Disease
  • Place of Presentation: Rehovot (Israel)
  • Year and Date: 2015-04-26
  • Int'l Joint Research / Invited
• [Presentation] Neuronal polarization in vivo: growing in a complex environment (2015)
  • Author(s): Kozo Kaibuchi
  • Organizer: Emerging concepts of the neuronal cytoskeleton (w15-46)
  • Place of Presentation: Puerto Varas, Chile
  • Year and Date: 2015-03-22 – 2015-03-26
  • Invited
• [Presentation] リン酸化セリン・スレオニン結合モチーフを生物学的フィルターとして用いたリン酸化プロテオミクス (2014)
  • Author(s): 西岡 朋生
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都府京都市左京区岩倉大鷺町、国立京都国際会館
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] Protein phosphorylation remains as a black box in signal transduction: developing a new method to search for substrates of protein kinases such as Rho-kinase. (2014)
  • Author(s): Kozo Kaibuchi
  • Organizer: 8th International Conference Inhibitors of Protein Kinases (IPK2014)
  • Place of Presentation: Warsaw, Poland
  • Year and Date: 2014-09-21 – 2014-09-25
  • Invited
• [Presentation] Systematic Substrate Screening of Protein Kinases using a Functional Proteomic Approach (2014)
  • Author(s): Mutsuki Amano
  • Organizer: 8th International Conference Inhibitors of Protein Kinases (IPK2014)
  • Place of Presentation: Warsaw, Poland
  • Year and Date: 2014-09-21 – 2014-09-25
• [Presentation] Molecular mechanisms regulating formation, polarity and migration in the developing brain (2014)
  • Author(s): Kozo Kaibuchi
  • Organizer: 12th Meeting of the Asian-Pacific Society for Neurochemistry (APSN)2014
  • Place of Presentation: Kaohsiung (Ta-Kao), Taiwan
  • Year and Date: 2014-08-23 – 2014-08-26
  • Invited
• [Presentation] Protein Phosporylation in Signal transduction (2014)
  • Author(s): Kozo Kaibuchi
  • Organizer: the 17th World Congress of Basic and Clinical Pharmacology (WCP2014)
  • Place of Presentation: Cape town, South Africa
  • Year and Date: 2014-07-13 – 2014-07-18
  • Invited
• [Presentation] Roles of the Rho Family Gtpases and Related Kinases in Polarized Migration (2014)
  • Author(s): Kozo Kaibuchi
  • Organizer: GRC (Gordon Research Conference) Signaling by Adhesion Receptors
  • Place of Presentation: ME(Maine), USA
  • Year and Date: 2014-06-22 – 2014-06-27
  • Invited
• [Presentation] Protein phosphorylation remains a black box in signal transduction: Developing a new method to search for substrates of polarity kinases including Rho-kinase (2014)
  • Author(s): Mutsuki Amano
  • Organizer: The 4th Asia Pacific Protein Association (APPA) Conference
  • Place of Presentation: Jeju, Korea
  • Year and Date: 2014-05-17 – 2014-05-20
  • Invited
• [Presentation] 神経関連キナーゼの基質の網羅的解析 (2013)
  • Author(s): 天野 睦紀
  • Organizer: 第13回日本蛋白質科学学会年会
  • Place of Presentation: とりぎん文化会館 （鳥取市）
• [Presentation] In vivo Screening of Kinase-Specific Substrates by Phosphatase Inhibitor and Kinase Inhibitor Substrate Screening (PIKISS) (2013)
  • Author(s): 西岡 朋生
  • Organizer: The 25th CDB Meeting Cilia and Centrosomes: from Fertilization to Cancer
  • Place of Presentation: 理研CDB （神戸市）
• [Presentation] 質量分析を用いたキナーゼ特異的基質探索法の開発 (2013)
  • Author(s): 西岡 朋生
  • Organizer: 第65回日本細胞生物学会大会
  • Place of Presentation: ウインクあいち （名古屋市）
• [Presentation] In vivo Screening of Kinase-Specific Substrates by Phosphatase Inhibitor and Kinase Inhibitor Substrate Screening (PIKISS) (2013)
  • Author(s): 西岡 朋生
  • Organizer: HUPO 12th Annual World Congress
  • Place of Presentation: パシフィコ横浜 （横浜市）
• [Presentation] Neuronal polarity in vitro and in vivo
  • Author(s): Kozo Kaibuchi
  • Organizer: International Symposium"Neocortical Organization 2"
  • Place of Presentation: 岡崎コンファレンスセンター （岡崎市）
  • Invited
• [Presentation] 蛋白質修飾によるシグナル伝達研究の新展開
  • Author(s): 貝淵 弘三
  • Organizer: 第13回日本蛋白質科学学会年会
  • Place of Presentation: とりぎん文化会館 （鳥取市）
  • Invited
• [Presentation] Neuronal polarity in vitro and in vivo
  • Author(s): Kozo Kaibuchi
  • Organizer: ASCB2013
  • Place of Presentation: New orleans, USA
  • Invited
• [Presentation] 神経関連キナーゼの基質の網羅的解析
  • Author(s): 天野 睦紀
  • Organizer: 第２７回日本医用マススペクトル学会東海支部講演会
  • Place of Presentation: 名古屋大学医学部 （名古屋市）
  • Invited