ゴルジ体を中心とするメンブレントラフィックの制御機構

• Project/Area Number: 15079204
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyoto University
• Principal Investigator: 中山 和久 Kyoto University, 薬学研究科, 教授 (40192679)
• Co-Investigator(Kenkyū-buntansha): 申 恵媛 (申 惠媛) 京都大学, 薬学研究科, 助教 (10345598)
• Project Period (FY): 2003 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥105,200,000 (Direct Cost: ¥105,200,000); Fiscal Year 2007: ¥18,400,000 (Direct Cost: ¥18,400,000); Fiscal Year 2006: ¥19,600,000 (Direct Cost: ¥19,600,000); Fiscal Year 2005: ¥19,600,000 (Direct Cost: ¥19,600,000); Fiscal Year 2004: ¥23,800,000 (Direct Cost: ¥23,800,000); Fiscal Year 2003: ¥23,800,000 (Direct Cost: ¥23,800,000)
• Keywords: ARF / Rab / メンブレントラフィック / 小胞輸送 / 細胞・組織 / 生体分子 / 蛋白質 / 薬学 / BIG2 / TGN / エンドソーム / GGA / Tom1 / GATドメイン / ユビキチン / X線結晶構造解析

Research Abstract

トランスゴルジ網(TGN)で機能する低分子量GTPaseのARL1とARFRP1、およびこれまでは主としてTGNに局在し、ARFのエフェクターであると考えられてきたArfaptin-1(POR2)とArfaptin-2(POR1)に関して以下のことを明らかにした。
1.これまで、ARL1はARFRP1の下流で機能し、TGNからの順行輸送とTGNへの逆行輸送の両方を調節すると考えられてきたが、RNA干渉法を用いた実験などによって、両者別個に機能し、ARFRP1はTGNからの順行輸送を、ARL1はエンドソームからTGNへの逆行輸送を調節することを明らかにしたこと。
2.Arfaptin-1とArfaptin-2のTGNへの局在が、ARFではなく、ARL1によって決定されることをRNA干渉法などによって明らかにしたこと。ただし、ARL1が関与することがこれまでに判明している輸送経路には関係しないこと。
3.タイムラプスイメージングで観察すると、Arfaptin-1とArfaptin-2はTGNから出て行くチューブル状の構造にダイナミックに存在すること。
一方、ゴルジ体やTGNで機能すると考えられていたARF3に関して、RNA干渉法を用いて以下のことを明らかにした。
4.ARF3は構造的に極めて類似するARF1とは異なり、主として初期エンドソームと後期エンドソームで機能し、リソソームでの分解経路を調節すること。

Research Products

• [Journal Article] Differential effects of depletion of ARL1 and ARFRP1 on membrane trafficking between the trans-Golgi network and endosomes (2009)
  • Author(s): Nishimoto-Morita, K., et al.
  • Journal Title: J.Biol.Chem. 284 Pages: 10583-10592
  • Peer Reviewed
• [Journal Article] Three homologous ArfGAPs participate in COPI-mediated transport (2009)
  • Author(s): Saitoh, A., et al.
  • Journal Title: J.Biol.Chem. 284(in press)
  • Peer Reviewed
• [Journal Article] Recruitment of Tom1 L1/Srcasm to endosomes and the midbody by Tsg101 (2008)
  • Author(s): Yanagida-Ishizaki, Y., et al.
  • Journal Title: Cell Struct.Funct. 33 Pages: 91-100
  • Peer Reviewed
• [Journal Article] Redundant roles of BIG2 and BIG1, guanine-nucleotide exchange factors for ARFs, in membrane traffic between the TGN and endosomes (2008)
  • Author(s): Ishizaki, R., et al.
  • Journal Title: Mol.Biol.Cell 19 Pages: 2650-2660
  • Peer Reviewed
• [Journal Article] f histidine decarboxylase through post-translational cleavageby caspase-9 in a mouse mastocytoma P-815. (2007)
  • Author(s): Furuta, K., Nakayama, K., Sugimoto, Y., Ichikawa, A. & Tanaka, S.
  • Journal Title: J. Biol. Chem. 282 Pages: 13438-13446
• [Journal Article] Membrane orientation of the precursor 74-kDa form of L-histidine decarboxylase. (2006)
  • Author(s): Furuta, K., Ichikawa, A., Nakayama, K. & Tanaka, S.
  • Journal Title: Inflamm. Res. 55 Pages: 185-191
• [Journal Article] Recruitment of clathrin onto endosomes by the Tom1-Tollip complex. (2006)
  • Author(s): Katoh, Y., Imakagura, H., Futatsumori, M. & Nakayama, K.
  • Journal Title: Biochem. Biophys. Res. Commun. 341 Pages: 143-149
• [Journal Article] AMY-1(associate or Myc-1)localization to the trans-Golgi network through interacting with BIG2, a guanine-nucleotide exchange factor for ADP-ribosylation factors (2006)
  • Author(s): Ishizaki, R., Shin, H.-W., Iguchi-Ariga, S. M. M., Ariga, H. & Nakayama, K
  • Journal Title: Genes Cells 11 Pages: 949-959
• [Journal Article] Monoubiquitylation of GGA3 by hVPS18 regulates its ubiquitin-binding ability. (2006)
  • Author(s): Yogosawa, S., Kawasaki, M., Wakatsuki, S., Kominami, E., Shiba, Y, Nakayama K, Kohsaka S, Akazawa C.
  • Journal Title: Biochem. Biophys. Res. Commun. 350 Pages: 82-90
• [Journal Article] Structural basis for Rabll-dependent membrane recruitment of FIP3/Arfophilin-1. (2006)
  • Author(s): Shiba, I., Koga, H., Shin, H.-W., Kawasaki, M., Kato, R., Nakayama, K. & Wakatsuki S.
  • Journal Title: Proc. Natl. Acad. Sci. USA 103 Pages: 15416-15421
• [Journal Article] Recruitment of clathrin onto endosomes by the Tom1-Tollip complex. (2006)
  • Author(s): Katoh, Y., Imakagura, H.Futatsumori, M., Nakayama, K.
  • Journal Title: Biochem.Biophys.Res.Commun. 341 Pages: 143-149
• [Journal Article] Structural basis for recognition of ubiquitinated cargo by Tom1-GAT domain (2005)
  • Author(s): Akutsu, M., Kawasaki, M., Katoh, Y., Shiba, T., Yamaguchi, Y., Kato, R., Kato, K., Nakayama, K., Wakatsuki, S.
  • Journal Title: FEBS Lett. 579 Pages: 5385-5391
• [Journal Article] Roles of ARFRP1 (ADP-ribosylation factor-related protein 1) in post-Golgi membrane trafficking. (2005)
  • Author(s): Shin, H.-W., Kobayashi, H., Kitamura, M., Waguri, S., Suganuma, T., Uchiyama, Y., Nakayama, K.
  • Journal Title: J.Cell Sci. 118 Pages: 4039-4048
• [Journal Article] Molecular mechanism of ubiquitin recognition by GGA3 GAT domain. (2005)
  • Author(s): Kawasaki, M., Shiba, T., Shiba, Y., Yamaguchi, Y., Matsugaki, N., Igarashi, N., Suzuki, M., Kato, R., Kato, K., Nakayama, K., Wakatsuki, S.
  • Journal Title: Genes Cells 10 Pages: 639-654
• [Journal Article] Structure determination of GGA-GAE and gl-ear in complex with peptides : crystallization of low-affinity complexes in membrane traffic. (2005)
  • Author(s): Yamada, Y., Inoue, M., Shiba, T., Kawasaki, M., Kato, R., Nakayama, K., Wakatsuki, S.
  • Journal Title: Acta.Cryst.D. 61 Pages: 731-736
• [Journal Article] Cellular and subcellular localization of EFA6C, a third member of the EFA6 family, in adult mouse Purkinje cells. (2005)
  • Author(s): Matsuya, S., Sakagami, H., Tohgo, A., Owada, Y., Shin, H.-W., Takeshima, H., Nakayama, K., Kokubun, S., Kondo, H.
  • Journal Title: J.Neurochem. 93 Pages: 674-685
• [Journal Article] Convergence of cell cycle regulation and growth factor signals on GRASP65. (2005)
  • Author(s): Yoshimura, S., Yoshioka, K., Barr, F.A., Lowe, M., Nakayama, K., Ohkuma, S., Nakamura, N.
  • Journal Title: J.Biol.Chem. 280 Pages: 23048-23056
• [Journal Article] メンブレントラフィックを調節する新規のクラスリンアダプターの構造と機能 (2005)
  • Author(s): 中山和久, 若槻壮市
  • Journal Title: 生化学 78 Pages: 1367-1381
• [Journal Article] Membrane recruitment of effector proteins by Arf and Rab GTPases. (2005)
  • Author(s): Kawasaki, M., Nakayama, K., Wakatsuki, S.
  • Journal Title: Curr.Opin.Struct.Biol. 15 Pages: 681-689
• [Journal Article] Tollip and Tom1 form a complex and recruit ubiquitin-conjugated proteins onto early endosomes. (2004)
  • Author(s): Katoh, Y., et al.
  • Journal Title: J.Biol.Chem. 279 Pages: 24435-24443
• [Journal Article] Stabilin-1 localises to endosomes and the trans-Golgi network in human macrophages and interacts with GGA adaptors. (2004)
  • Author(s): Kzhyshkowska, J., et al.
  • Journal Title: J. Leukocyte Biol. 76 Pages: 1151-1161
• [Journal Article] BIG2,a guanine nucleotide exchange factor for ADP- ribosylation factors : its localization to recycling endosomes and implication in the endosome integrity. (2004)
  • Author(s): Shin, H.-W., et al.
  • Journal Title: Mol.Biol. Cell 15 Pages: 5283-5294
• [Journal Article] Guanine nucleotide exchange factors for Arf GTPases : their diverse functions in membrane traffic. (2004)
  • Author(s): Shin, H.-W., Nakayama, K.
  • Journal Title: J.Biochem. 136 Pages: 761-767
  • NAID: 10016204135
• [Presentation] Arfのグアニンヌクレオチド交換因子BIG1とBIG2のトランスゴルジ網への局在機構の解析 (2008)
  • Author(s): 高島皓平ら
  • Organizer: 第31回日本分子生物学会年会・第81回日本生化学会大会合同大会シンポジウム
  • Place of Presentation: 神戸ポートピアホテル
  • Year and Date: 2008-12-10
• [Publications] Shiba, Y., et al.: "GAT(GGA and Tom1) domain responsible for ubiquitin binding and ubiquitination"J.Biol.Chem.. 279. 7105-7111 (2004)
• [Publications] Shakoori, A., et al.: "Identification of a five-pass transmembrane protein family localizing in the Golgi apparatus and the ER"Biochem.Biophys.Res.Commun.. 312. 850-857 (2003)
• [Publications] Matsukawa, J., et al.: "Role of ADP-ribosylation factor 6 (ARF6) in gastric acid secretion"J.Biol.Chem.. 278. 36470-36475 (2003)
• [Publications] Wakasugi, M., et al.: "Predominant expression of the short form of GGA3 in human cell lines and tissues"Biochem.Biophys.Res.Commun.. 306. 687-692 (2003)
• [Publications] Shiba, T., et al.: "Molecular mechanism of membrane recruitment of GGA by ARF in lysosomal protein transport"Nat.Struct.Biol.. 10. 368-393 (2003)