環境ストレス応答におけるポストゴルジオルガネラのダイナミクスと生理的意義の解明

• Project Area: Environmental sensing of plants: Signal perception, processing and cellular responses
• Project/Area Number: 25120706
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 植村 知博 東京大学, 理学(系)研究科(研究院), 助教 (90415092)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥15,080,000 (Direct Cost: ¥11,600,000、Indirect Cost: ¥3,480,000); Fiscal Year 2014: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000); Fiscal Year 2013: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
• Keywords: オルガネラ / 環境ストレス / イメージング / SNARE / シロイヌナズナ / 膜交通 / TGN

Outline of Annual Research Achievements

トランスゴルジ網（TGN）には、ゴルジ体のトランス槽側に存在するGA-TGN (Golgi-associated TGN)とゴルジ体とは独立して存在するGI-TGN (Golgi-released independent TGN)の２種類が存在する。本年度は、うどんこ病菌応答におけるGI-TGNのダイナミクスを観察した。その結果、うどんこ病菌に対する抵抗応答時において、GI-TGNの数が増加することを発見した。抗菌物質の分泌のために分泌経路が活性化されことに伴い、GI-TGNもその動態を大きく変化させると考えている。また、当研究室で開発された超解像ライブイメージン顕微鏡（SCLIM: super-resolution confocal live imaging microscopy）を用いてトランスゴルジ網（TGN）の観察を行った結果、TGNに局在するタンパク質は一様に存在しているのではなく、ドメイン構造を有していることが示唆された。
次に塩ストレス応答におけるオルガネラダイナミクスの観察を行った。その結果、高塩ストレス下ではTGNおよび単膜系オルガネラが凝集体を形成することを見出した。また、TGN機能が損なわれたsyp4変異体ではNHX6の細胞内局在が異常になることから、syp4変異体が高塩ストレスに弱くなる原因の一つとしてNHX6が正しく局在できないために細胞内の塩濃度を調整できないことが考えられた。
ストレス環境下でのポストゴルジオルガネラのダイナミクスをライブイメージングにより明らかにするための観察システムの構築を継続して行った。高速撮影が可能なニポウディスク方式の倒立共焦点レーザー顕微境に多点画像取得機能と自動フォーカス機能を搭載したシステムを構築したが、根などの厚みのあるサンプルでの長時間イメージンについてはまだ良い結果が得られておらず現在改良中である。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Dynamic behavior of the trans-Golgi network in root tissues of Arabidopsis revealed by super-resolution live imaging (2014)
  • Author(s): T. Uemura, Y. Suda, T. Ueda, and A. Nakano
  • Journal Title: Plant Cell Physiol Volume: 55 Issue: 4 Pages: 694-670
  • DOI: 10.1093/pcp/pcu010
  • Peer Reviewed
• [Journal Article] A unique HEAT repeat-containing protein SHOOT GRAVITROPISM6 is involved in vacuolar membrane dynamics in gravity sensing cells of Arabidopsis inflorescence stem. (2014)
  • Author(s): 2.Hashiguchi, Y., Yano, D., Nagafusa, K., Kato, T., Saito, C., Uemura, T., Ueda, T., Nakano, A., Tasaka, M and * Morita, MT
  • Journal Title: Plant Cell Physiol Volume: 55 Pages: 811-822
  • Peer Reviewed
• [Journal Article] The ESCRT-III interacting deubiquitinating enzyme AMSH3 is essential for degradation of ubiquitinated membrane proteins in Arabidopsis thaliana (2014)
  • Author(s): 3.Katsiarimpa, A., Muñoz, A., Kalinowska, K., Uemura, T., Rojo, E. and Isono, E
  • Journal Title: Plant Cell Physiol Volume: 55 Pages: 727-736
  • Peer Reviewed
• [Journal Article] Interactomics of Qa-SNARE in Arabidopsis thaliana (2014)
  • Author(s): Fujiwara, M., Uemura, T., Ebine, K., Nishimori, Y., Ueda, T., Nakano, A., Sato, MH. and Fukao, Y.
  • Journal Title: Plant Cell Phys. Volume: 55 Issue: 4 Pages: 781-789
  • DOI: 10.1093/pcp/pcu038
  • Peer Reviewed / Open Access
• [Journal Article] Sex-specific post-translational regulation of the gamete fusogen GCSI in the isogamous volvocine alga Gonium pectorale. (2014)
  • Author(s): Kawai-Toyooka H, Mori T, Hamaji T, Suzuki M, Olson BJ, Uemura T, Ueda T, Nakano A, Toyoda A, Fujiyama A, Nozaki H
  • Journal Title: EUkaryotic Cell Volume: (in press) Issue: 5 Pages: 648-656
  • DOI: 10.1128/ec.00330-13
  • Peer Reviewed
• [Journal Article] Plant vacuolar trafficking occurs through distinctly regulated pathways (2014)
  • Author(s): 6.Ebine, K., Inoue, T., Ito, J., Ito, E., Uemura, T., Goh, T., Abe, H., Sato, K., Nakano, A. and Ueda, T
  • Journal Title: Curr. Biol Volume: 24 Pages: 1375-1382
  • Peer Reviewed
• [Journal Article] The powdery mildew resistance protein RPW8.2 is carried on VAMP721/722 vesicles to the extrahaustorial membrane of haustorial complexes (2014)
  • Author(s): 7.Kim, H., O'Connell, R., Maekawa-Yoshikawa, M., Uemura, T., Neumann, U., Schulze-Lefert P
  • Journal Title: Plant J. Volume: 79 Issue: 5 Pages: 835-847
  • DOI: 10.1111/tpj.12591
  • Peer Reviewed
• [Journal Article] Formation and Maintenance of the Golgi Apparatus in Plant Cells (2014)
  • Author(s): Yoko Ito, Tomohiro Uemura, and Akihiko Nakano
  • Journal Title: International Review of Cell & Molecular Biology Volume: 310 Pages: 221-287
  • DOI: 10.1016/b978-0-12-800180-6.00006-2
  • ISBN: 9780128001806
  • Peer Reviewed
• [Journal Article] Plant vacuolar trafficking driven by RAB and SNARE proteins (2014)
  • Author(s): Uemura, T. and Ueda, T.
  • Journal Title: Curr. Opin. Plant Biol. Volume: 22 Pages: 116-121
  • DOI: 10.1016/j.pbi.2014.10.002
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A unique HEAT repeat-containing protein SHOOT GRAVITROPISM 6 is involved in vacuolar membrane dynamics in gravity sensing cells of Arabidopsis inflorescence stem (2014)
  • Author(s): Hashiguchi, Y., Yano, D., Nagafusa, K., Kato, T., Saito, C., Uemura, T., Ueda, T., Nakano, A., Tasaka, M. and Morita, MT
  • Journal Title: Plant Cell Phys. Volume: 55 Issue: 4 Pages: 811-822
  • DOI: 10.1093/pcp/pcu020
  • Peer Reviewed
• [Journal Article] The ESCRT-III interacting deubiquitinating enzyme AMSH3 is essential for degradation of ubiquitinated membrane proteins in Arabidopsis thaliana. (2014)
  • Author(s): Katsiarimpa, A., Muñoz, A., Kalinowska, K., Uemura, T., Rojo, E. and Isono, E.
  • Journal Title: Plant Cell Physiol. Volume: 55 Issue: 4 Pages: 727-736
  • DOI: 10.1093/pcp/pcu019
  • Peer Reviewed
• [Journal Article] Arabidopsis RABA1 GTPases are involved in transport between the trans-Golgi network and the plasma membrane, and are required for salinity stress tolerance. (2013)
  • Author(s): Rin Asaoka, Tomohiro Uemura, Jun Ito, Masaru Fujimoto, Emi Ito, Takashi Ueda, Akihiko Nakano
  • Journal Title: The Plant Journal Volume: 73 Issue: 2 Pages: 240-249
  • DOI: 10.1111/tpj.12023
  • Peer Reviewed
• [Journal Article] Physical interaction of noral organs contr01S petal morphogenesiS in Arabidopsis thaliana. (2013)
  • Author(s): Seiji Takeda, et al.
  • Journal Title: Plant Physiol. Volume: 61 Issue: 3 Pages: 1242-1250
  • DOI: 10.1104/pp.112.212084
  • Peer Reviewed
• [Journal Article] RABA Members Act in Distinct Steps of Subcellular Trafficking of the FLAGELLIN SENSING2 Receptor. (2013)
  • Author(s): Choi, et al.
  • Journal Title: Plant Cell Volume: (in press) Issue: 3 Pages: 1174-1187
  • DOI: 10.1105/tpc.112.108803
  • Peer Reviewed
• [Journal Article] Cell polarity and patterning by PIN trafficking through early endosomal compartments in Arabidopsis thaliana. (2013)
  • Author(s): Tanaka H, Kitakura S, Rakusová H , Uemura T, Feraru MI, De Rycke R, Robert S, Kakimoto T, Friml J.
  • Journal Title: PLoS Genet. Volume: 9 Issue: 5 Pages: e1003540-e1003540
  • DOI: 10.1371/journal.pgen.1003540
  • Peer Reviewed
• [Journal Article] New insights into the role of Arabidopsis RABA1 GTPases in salinity stress tolerance. (2013)
  • Author(s): Asaoka Rin, Uemura Tomohiro, Nishida Sho, Fujiwara Toru, Ueda Takashi, Nakano Akihiko
  • Journal Title: Plant Signal and Behavior Volume: 8 Issue: 9 Pages: e25377-e25377
  • DOI: 10.4161/psb.25377
  • Peer Reviewed / Open Access
• [Journal Article] Plant TGNs: dynamics and physiological functions (2013)
  • Author(s): Tomohiro Uemura and Akihiko Nakano
  • Journal Title: Histochem. Cell Biol. Volume: 140 Issue: 3 Pages: 341-345
  • DOI: 10.1007/s00418-013-1116-7
  • Peer Reviewed
• [Presentation] ポストゴルジオルガネラが担う環境ストレス応答 (2015)
  • Author(s): 植村知博
  • Organizer: 第17回植物オルガネラワークショップ
  • Place of Presentation: 東京
  • Year and Date: 2015-03-15
  • Invited
• [Presentation] 植物におけるトランスゴルジネットワーク（ＴＧＮ）の動態 (2014)
  • Author(s): 植村知博
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-27
  • Invited
• [Presentation] The dynamics of trans-Golgi network (TGN) in plants. (2014)
  • Author(s): Tomohiro Uemura and Akihiko Nakano.
  • Organizer: 38th Naito Conference on Molecule-based biological systems
  • Place of Presentation: SAPPORO, Japan
  • Year and Date: 2014-10-07 – 2014-10-10
• [Presentation] 植物におけるポストゴルジオルガネラの動態と生理機能の研究 (2014)
  • Author(s): 植村知博
  • Organizer: 日本植物学会第78回大会
  • Place of Presentation: 川崎
  • Year and Date: 2014-09-13
  • Invited
• [Presentation] The dynamics of trans-Golgi network (TGN) in plants. (2014)
  • Author(s): Tomohiro Uemura, Takashi Ueda, Akihiko Nakano
  • Organizer: 17th European Network for plant Endomembrane Research Meeting
  • Place of Presentation: Lecce, Italy
  • Year and Date: 2014-09-08 – 2014-09-11
• [Presentation] The dynamics and functions of Qa-SNAREs localized on the trans-Golgi network on response to Colletotrichum gloeosporioides. (2014)
  • Author(s): Tomohiro Uemura, Takashi Shimada, Takashi Ueda, Akihiko Nakano.
  • Organizer: IS-MPMI 2014 XVI International Congress
  • Place of Presentation: Rhodes, Greece
  • Year and Date: 2014-07-06 – 2014-07-10
• [Presentation] The dynamics of trans-Golgi network (TGN) in plants (2014)
  • Author(s): Tomohiro Uemura, Yasuyuki Suda, Takashi Ueda and Akihiko Nakano.
  • Organizer: 第62回日本細胞生物学会大会
  • Place of Presentation: 奈良
  • Year and Date: 2014-06-12
• [Presentation] The dynamics of trans-Golgi network (TGN) in plants. (2013)
  • Author(s): Tomohiro Uemura
  • Organizer: Golgi Apparatus Symposium
  • Place of Presentation: Bad Ischl, Austria
  • Invited