Regulation of vesicular trafficking by tethering complex and SNARE proteins

• Project/Area Number: 15H04370
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Cell biology
• Research Institution: Kyoto University
• Principal Investigator: Nakayama Kazuhisa 京都大学, 薬学研究科, 教授 (40192679)
• Co-Investigator(Renkei-kenkyūsha): KATOH Yohei 京都大学, 大学院薬学研究科, 助教 (90568172)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000); Fiscal Year 2017: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2016: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2015: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
• Keywords: 小胞輸送 / SNAREタンパク質 / Exocyst複合体 / IFT-B複合体 / SNARE / Exorcist / VIPアッセイ / IFT複合体 / 細胞・組織 / 細胞内タンパク質輸送 / タンパク質複合体

Outline of Final Research Achievements

Membrane fusion, which is the last step of vesicular trafficking, is mediated by SNARE proteins. In this study, we have revealed that fusion of recycling vesicles containing transferrin-transferrin receptor is regulated by t-SNARE proteins, SNAP-23/SNAP-25 in conjunction with the Exocyst tethering complex.
By taking advantage of the visible immunoprecipitation (VIP) assay, we have revealed the architectures of various protein complex, including the Exocyst complex, involved in protein trafficking within cells.

Research Products

• [Journal Article] Ciliopathy-associated mutations of IFT122 impair ciliary protein trafficking but not ciliogenesis (2018)
  • Author(s): Takahara, M., Katoh, Y., Nakamura, K., Hirano, T., Sugaga, M., Tsurumi, Y. Nakayama, K.
  • Journal Title: Hum. Mol. Genet. Volume: 27 Issue: 3 Pages: 516-528
  • DOI: 10.1093/hmg/ddx421
  • Peer Reviewed
• [Journal Article] BBS1 is involved in retrograde trafficking of ciliary GPCRs in the context of the BBSome complex (2018)
  • Author(s): Nozaki, S., Katoh, Y., Kobayashi, T., Nakayama, K.
  • Journal Title: PLOS ONE Volume: 13 Issue: 3 Pages: 1-17
  • DOI: 10.1371/journal.pone.0195005
  • NAID: 120006466602
  • Peer Reviewed / Open Access
• [Journal Article] Phospholipid-flipping activity of P4-ATPase drives membrane curvature. (2018)
  • Author(s): Takada, N., Naito, T., Inoue, T., Nakayama, K., Takatsu, H., and *Shin, H.-W.
  • Journal Title: The EMBO Journal Volume: in press Issue: 9
  • DOI: 10.15252/embj.201797705
  • NAID: 120007144982
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Practical method for targeted disruption of cilia-related genes by using CRISPR/Cas9-mediated, homology-independent knock-in system (2017)
  • Author(s): Katoh Y, Michisaka S, Nozaki S, Funabashi T, Hirano T, Takei R, Nakayama K.
  • Journal Title: Mol. Biol. Cell Volume: 28 Issue: 7 Pages: 898-906
  • DOI: 10.1091/mbc.e17-01-0051
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] RABL2 interacts with the intraflagellar transport-B complex and CEP19 and participates in ciliary assembly (2017)
  • Author(s): Nishijima Yuya、Hagiya Yohei、Kubo Tomohiro、Takei Ryota、Katoh Yohei、Nakayama Kazuhisa
  • Journal Title: Molecular Biology of the Cell Volume: 28 Issue: 12 Pages: 1652-1666
  • DOI: 10.1091/mbc.e17-01-0017
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Phospholipid flippase ATP11C is endocytosed and downregulated following Ca²⁺-mediated protein kinase C activation. (2017)
  • Author(s): Takatsu H, Takayama M, Naito T, Takada N, Tsumagari K, Ishihama Y, Nakayama K, Shin HW.
  • Journal Title: Nat Commun. Volume: Nov 10;8(1) Issue: 1 Pages: 1423-1423
  • DOI: 10.1038/s41467-017-01338-1
  • NAID: 120006365525
  • Peer Reviewed / Open Access
• [Journal Article] Regulation of ciliary retrograde protein trafficking by the Joubert syndrome proteins ARL13B and INPP5E. (2017)
  • Author(s): Nozaki S, Katoh Y, Terada M, Michisaka S, Funabashi T, Takahashi S, Kontani K, Nakayama K.
  • Journal Title: J. Cell. Sci. Volume: 130 Issue: 3 Pages: 563-576
  • DOI: 10.1242/jcs.197004
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Intraflagellar transport-A complex mediates ciliary entry and retrograde trafficking of ciliary G protein-coupled receptors (2017)
  • Author(s): Hirano T, Katoh Y, Nakayama K
  • Journal Title: Mol. Biol. Cell Volume: 28 Issue: 3 Pages: 429-439
  • DOI: 10.1091/mbc.e16-11-0813
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Ciliary entry of KIF17 is dependent on its binding to the IFT-B complex via IFT46-IFT56 as well as on its nuclear localization signal (2017)
  • Author(s): Funabashi T, Katoh Y, Michisaka S, Terada M, Sugawa M, Nakayama K
  • Journal Title: Mol. Biol. Cell Volume: 28 Issue: 5 Pages: 624-633
  • DOI: 10.1091/mbc.e16-09-0648
  • NAID: 120005997941
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Modulation of primary cilia length by melanin-concentrating hormone receptor 1. Cell Signaling (2016)
  • Author(s): Hamamoto A, Yamato S, Katoh Y, Nakayama K, Yoshimura K, Takeda S, Kobayashi Y, *Saito Y
  • Journal Title: Cellular Signalling Volume: 28 Issue: 6 Pages: 272-584
  • DOI: 10.1016/j.cellsig.2016.02.018
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Overall architecture of the intraflagellar transport (IFT)-B complex containing Cluap1/IFT38 as an essential component of the IFT-B peripheral subcomplex. (2016)
  • Author(s): Katoh Y, Terada M, Nishijima Y, Takei R, Nozaki S, Hamada H, Nakayama K
  • Journal Title: The Journal of Biological Chemistry Volume: 291 Issue: 21 Pages: 10962-10975
  • DOI: 10.1074/jbc.m116.713883
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Alteration of transbilayer phospholipid compositions is involved in cell adhesion, cell spreading, and focal adhesion formation (2016)
  • Author(s): Miyano R, Matsumoto T, Takatsu H, Nakayama K, Shin HW
  • Journal Title: FEBS Lett. Volume: 590 Issue: 14 Pages: 2138-2145
  • DOI: 10.1002/1873-3468.12247
  • NAID: 120007159976
  • Peer Reviewed
• [Journal Article] The phospholipid flippase ATP9A is required for the recycling pathway from the endosomes to the plasma membrane (2016)
  • Author(s): Tanaka Y, Ono N, Shima T, Tanaka G, Katoh Y, Nakayama K, Takatsu H, Shin HW
  • Journal Title: Mol. Biol. Cell Volume: 27 Issue: 24 Pages: 3883-3893
  • DOI: 10.1091/mbc.e16-08-0586
  • NAID: 120005997942
  • Peer Reviewed / Open Access
• [Journal Article] Class I Arfs (Arf1 and Arf3) and Arf6 are localized to the Flemming body and play important roles in cytokinesis. (2016)
  • Author(s): Hanai, A., Ohgi, M., Yagi, C., Ueda, T., Shin, H.-W., and Nakayama, K.
  • Journal Title: Journal of Biochemistry Volume: 159 Issue: 2 Pages: 201-208
  • DOI: 10.1093/jb/mvv088
  • Peer Reviewed
• [Journal Article] GM130 is a parallel tetramer with a flexible rod-like structure and N-terminally open (Y-shaped) and closed (I-shaped) conformations. (2015)
  • Author(s): Ishida R, Yamamoto A, Nakayama K, Sohda M, Misumi Y, Yasunaga T, Nakamura N.
  • Journal Title: FEBS J. Volume: 印刷中 Issue: 11 Pages: 2232-2244
  • DOI: 10.1111/febs.13271
  • Peer Reviewed
• [Journal Article] Architectures of multisubunit complexes revealed by a visible immunoprecipitation assay using fluorescent fusion proteins. (2015)
  • Author(s): Katoh Y, Nozaki S, Hartanto D, Miyano R, Nakayama K
  • Journal Title: Journal of Cell Science Volume: 128 Issue: 12 Pages: 2351-2362
  • DOI: 10.1242/jcs.168740
  • Peer Reviewed / Open Access
• [Journal Article] Phospholipid flippase ATP10A translocates phosphatidylcholine and is involved in plasma membrane dynamics. (2015)
  • Author(s): Naito, T., Takatsu, H., Miyano, R., Takada, N., Nakayama, K., and Shin, H.-W.
  • Journal Title: Journal of Biological Chemistry Volume: 290 Issue: 24 Pages: 15004-15017
  • DOI: 10.1074/jbc.m115.655191
  • NAID: 120005690225
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] SNAP23/25 and VAMP2 mediate exocytic event of transferrin receptor-containing recycling vesicles. (2015)
  • Author(s): Kubo K, Kobayashi M, Nozaki S, Yagi C, Hatsuzawa K, Katoh Y, Shin HW, Takahashi S, Nakayama K
  • Journal Title: Biology Open Volume: 4 Issue: 7 Pages: 910-920
  • DOI: 10.1242/bio.012146
  • NAID: 120005997718
  • Peer Reviewed / Open Access
• [Journal Article] COPI-mediated retrieval of SCAP is critical for regulating lipogenesis under basal and sterol-deficient conditions. (2015)
  • Author(s): Takashima, K., Saitoh, A., Funabashi, T., Hirose, S., Yagi, C., Nozaki, S., Sato, R., Shin, H.-W., and Nakayama, K.
  • Journal Title: Journal of Cell Science Volume: 128 Pages: 2805-2815
  • DOI: 10.1242/jcs.164137
  • Peer Reviewed
• [Journal Article] ATP11C mutation is responsible for the defect in phosphatidylserine uptake in UPS-1 cells. (2015)
  • Author(s): Takada, N., Takatsu, H., Miyano, R., Nakayama, K., and Shin, H.-W.
  • Journal Title: Jounal of Lipid Research Volume: 56 Issue: 11 Pages: 2151-2157
  • DOI: 10.1194/jlr.m062547
  • NAID: 120005707794
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] IFT-A複合体によるGPCRの繊毛内移行と繊毛内逆行輸送の調節 (2017)
  • Author(s): 加藤洋平、平野友章、中山和久
  • Organizer: 第64回日本生化学会近畿支部例会
• [Presentation] ダイニン2複合体の構築様式および繊毛内タンパク質輸送における役割の解明 (2017)
  • Author(s): 鶴見侑大、濵田勇輝、 加藤洋平、中山和久
  • Organizer: 第64回日本生化学会近畿支部例会
• [Presentation] Joubert症候群原因遺伝子産物ARL13BとINPP5Eによるタンパク質の繊毛内逆行輸送の制御 (2017)
  • Author(s): 野崎梢平、加藤洋平、中山和久
  • Organizer: 第69回日本細胞生物学会大会
• [Presentation] IFT-B複合体と相互作用するキネシン２による繊毛形成の調節 (2017)
  • Author(s): 船橋輝記、加藤洋平、中山和久
  • Organizer: 第69回日本細胞生物学会大会
• [Presentation] RABL2はIFT-B複合体およびCEP19との相互作用を介して繊毛形成を調節する (2017)
  • Author(s): 西島侑哉、萩谷遥平、久保智広、武井領汰、加藤洋平、中山和久
  • Organizer: 第69回日本細胞生物学会大会
• [Presentation] ダイニン2複合体の構築様式およびIFT複合体との相互作用の解明 (2017)
  • Author(s): 濵田勇輝、鶴見侑大、加藤洋平、中山和久
  • Organizer: 第69回日本細胞生物学会大会
• [Presentation] 繊毛内タンパク質輸送複合体の構築様式と機能 (2017)
  • Author(s): 中山和久
  • Organizer: 第41回蛋白質の酵素の構造と機能に関する九州シンポジウム
  • Invited
• [Presentation] IFT-B複合体に依存するBBSome複合体の繊毛内移行 (2017)
  • Author(s): 野崎梢平、加藤洋平、中山和久
  • Organizer: ConBio2017．2017年度生命科学系学会合同年次大会
• [Presentation] 膨張顕微鏡法を用いた一次繊毛の超解像イメージング (2017)
  • Author(s): 加藤洋平、千葉秀平、中山和久
  • Organizer: ConBio2017．2017年度生命科学系学会合同年次大会
• [Presentation] IFT-B複合体のサブユニットIFT70の機能に関する研究 (2017)
  • Author(s): 武井領汰、加藤洋平、中山和久
  • Organizer: ConBio2017．2017年度生命科学系学会合同年次大会
• [Presentation] ダイニン2複合体の構築様式およびIFT複合体との相互作用の解明 (2017)
  • Author(s): 濱田勇輝、鶴見侑大、加藤洋平、中山和久
  • Organizer: ConBio2017．2017年度生命科学系学会合同年次大会
• [Presentation] リン脂質フリップ活性による膜曲率の形成 (2017)
  • Author(s): 高田直人、内藤明樹、井上尊生、中山和久、高津宏之、申惠媛
  • Organizer: ConBio2017．2017年度生命科学系学会合同年次大会
• [Presentation] 脂質フリッパーゼの新奇基質としてグルコシルセラミドの発見とその輸送メカニズム (2017)
  • Author(s): 内藤朋樹、Bartholomew Roland、高津宏之、中山和久、Todd Graham、申惠媛
  • Organizer: ConBio2017．2017年度生命科学系学会合同年次大会
• [Presentation] リン脂質フリッパーゼATP11Cのスプライシングバリアント間の機能的相違 (2017)
  • Author(s): 高山真裕、高津宏之、中山和久、申惠媛
  • Organizer: ConBio2017．2017年度生命科学系学会合同年次大会
• [Presentation] 非アポトーシス細胞におけるリン脂質フリッパーゼATP11Cの制御機構 (2017)
  • Author(s): 高津宏之、高山真裕、中山和久、申惠媛
  • Organizer: ConBio2017．2017年度生命科学系学会合同年次大会
• [Presentation] 「観るだけでわかるタンパク質間相互作用解析法」の開発と繊毛内タンパク質輸送複合体 IFT-B の構築様式の解明 (2016)
  • Author(s): 加藤洋平、寺田将也、中山和久
  • Organizer: 第63回日本生化学会近畿支部例会
• [Presentation] KIF17 と IFT 複合体の相互作用様式の解明と繊毛内輸送における役割． (2016)
  • Author(s): 船橋輝記、加藤洋平、中山和久
  • Organizer: 第68回日本細胞生物学会大会
• [Presentation] 繊毛内タンパク質輸送複合体IFT-Aの構築様式と機能の解明． (2016)
  • Author(s): 平野友章、加藤洋平、中山和久
  • Organizer: 第68回日本細胞生物学会大会
• [Presentation] 繊毛内のイノシトールリン脂質代謝とタンパク質の局在におけるArl13b とIFT-B の役割 (2016)
  • Author(s): 野崎梢平、加藤洋平、中山和久
  • Organizer: 第68回日本細胞生物学会大会
• [Presentation] 観るだけでわかるタンパク質間相互作用解析法」を活用した繊毛内タンパク質輸送複合体IFT-Bの構築様式の解明 (2016)
  • Author(s): 加藤洋平、寺田将也、野崎梢平、中山和久
  • Organizer: 第68回日本細胞生物学会大会
• [Presentation] Trafficking Machineries within Cilia: Architectures and Functions of the IFT-A and IFT-B complexes (2016)
  • Author(s): Nakayama, K., Katoh, Y., Terada, T., Nozaki, S., Hirano, T. & Funabashi, T.
  • Organizer: 第68回日本細胞生物学会大会
  • Invited
• [Presentation] IFT-A複合体と相互作用するC11orf74の繊毛内タンパク質輸送における役割 (2016)
  • Author(s): 高原万梨子、平野友章、加藤洋平、中山和久
  • Organizer: 第89回日本生化学会大会
• [Presentation] キネシン2とIFT複合体の相互作用の様式の解明と繊毛形成における役割 (2016)
  • Author(s): 船橋輝記、加藤洋平、中山和久
  • Organizer: 第89回日本生化学会大会
• [Presentation] P4-ATPaseによるリン脂質フリップが膜変形に与える影響 (2016)
  • Author(s): 高田直人、中山和久、申 惠媛
  • Organizer: 第89回日本生化学会大会
• [Presentation] リン脂質フリッパーゼATP11CのPKCによる制御機構 (2016)
  • Author(s): 高津宏之、高山真裕、中山和久、申 惠媛
  • Organizer: 第89回日本生化学会大会
• [Presentation] ジュベール症候群原因遺伝子産物Arl13bによって制御される繊毛内タンパク質輸送の分子機構 (2016)
  • Author(s): 加藤洋平、野崎梢平、中山和久
  • Organizer: 第89回日本生化学会大会
  • Invited
• [Presentation] Phospholipid flippase ATP10A, a member of type IV P-type ATPases, translocates phosphatidylcholine and is involved in plasma membrane dynamics (2016)
  • Author(s): Naito, T., Takatsu, H., Miyano, R., Takada, N., Nakayama, K. & Shin, H.-W.
  • Organizer: 2016 ASCB Annual Meeting
  • Int'l Joint Research
• [Presentation] Regulation of ciliary retrograde protein trafficking by Joubert syndrome proteins Arl13b and INPP5E (2016)
  • Author(s): Nozaki, S., Katoh, Y. & Nakayama, K.
  • Organizer: 2016 ASCB Annual Meeting
  • Int'l Joint Research
• [Presentation] ホスファチジルセリン取り込み異常のCHO-K1変異株（UPS-1）におけるflippaseの発現解析． (2015)
  • Author(s): 高田直人、高津宏之、宮野吏永、中山和久、申惠媛
  • 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大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] 繊毛内タンパク質輸送複合体IFT-Bの構築様式とCLUAP1の機能の解明． (2015)
  • Author(s): 寺田将也、加藤洋平、野崎梢平、武井領汰、中山和久
  • 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大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] 中心体/基底小体で複合体を形成するRabL2とCep19の機能の解析． (2015)
  • Author(s): 西島侑哉、萩谷遥平、加藤洋平、中山和久
  • 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大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] COPI被覆小胞を介するSCAP-SREBP複合体のゴルジ体から小胞体への逆行輸送による脂質代謝の調節． (2015)
  • Author(s): 船橋輝記、高島皓平、齋藤明奈、廣瀬祥平、申惠媛、中山和久
  • 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大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] Visible immunoprecipitation（VIP）アッセイによるExocyst複合体の構築様式の解明． (2015)
  • Author(s): 野崎梢平、加藤洋平、中山和久
  • Organizer: 第14回 次世代を担う若手ファーマ・バイオフォーラム2015
  • Place of Presentation: 千葉
  • Year and Date: 2015-09-12
• [Presentation] COPI被覆小胞を介するSCAP-SREBP複合体のゴルジ体-小胞体間の逆行輸送による脂質代謝調節． (2015)
  • Author(s): 船橋輝記、高島皓平、齋藤明奈、廣瀬祥平、申 惠媛、中山和久
  • Organizer: 第14回 次世代を担う若手ファーマ・バイオフォーラム2015
  • Place of Presentation: 千葉
  • Year and Date: 2015-09-12
• [Presentation] リン脂質フリッパーゼATP10AのPCフリップ活性および細胞膜ダイナ ミクスへの関与． (2015)
  • Author(s): 内藤朋樹、高津宏之、宮野吏永、高田直人、中山和久、申 惠媛
  • Organizer: 第14回 次世代を担う若手ファーマ・バイオフォーラム2015
  • Place of Presentation: 千葉
  • Year and Date: 2015-09-12
• [Presentation] Architectures of the BBSome, IFT-B, and Exocyst Complexes Revealed by Visible Immunoprecipitation (VIP) Assay Using Fluorescent Fusion Proteins. (2015)
  • Author(s): Nakayama, K., Katoh, Y., Nozaki, S., & Terada, M.
  • Organizer: FASEB Science Research Conference, “Biology o f Cilia and Flagella”
  • Place of Presentation: Snowmass Village, Colorado,
  • Year and Date: 2015-07-19
  • Int'l Joint Research
• [Presentation] Visible immunoprecipitation (VIP) アッセイによるマルチサブユニット複合体の構築様式の解明 (2015)
  • Author(s): 加藤洋平、野崎梢平、寺田将也、中山和久
  • Organizer: 第67回日本細胞生物学会大会
  • Place of Presentation: 東京
  • Year and Date: 2015-06-30
• [Presentation] リン脂質フリッパーゼATP10AのPCフリップ活性及び細胞膜ダイナミクスへの関与． (2015)
  • Author(s): 内藤朋樹、高津宏之、宮野吏永、高田直人、中山和久、申惠媛
  • Organizer: 第62回日本生化学会近畿支部例会
  • Place of Presentation: 草津
  • Year and Date: 2015-05-16