細胞移動先端におけるアクチン細胞骨格制御機構の解明

• Project/Area Number: 17687020
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Cell biology
• Research Institution: The University of Tokyo
• Principal Investigator: 末次 志郎 The University of Tokyo, 分子細胞生物学研究所, 講師 (70345031)
• Project Period (FY): 2005 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥30,290,000 (Direct Cost: ¥23,300,000、Indirect Cost: ¥6,990,000); Fiscal Year 2007: ¥9,880,000 (Direct Cost: ¥7,600,000、Indirect Cost: ¥2,280,000); Fiscal Year 2006: ¥9,880,000 (Direct Cost: ¥7,600,000、Indirect Cost: ¥2,280,000); Fiscal Year 2005: ¥10,530,000 (Direct Cost: ¥8,100,000、Indirect Cost: ¥2,430,000)
• Keywords: がん / 細胞・組織 / シグナル伝達 / 脂質 / タンパク質 / IRSp53 / WAVE2 / Arp2 / 3 / 膜 / アクチン重合 / アクチン / 3複合体 / WAVE / 仮足形成 / ラメリポディア

Research Abstract

細胞の移動は、多細胞生物に見られる様々な細胞機能の理解に必須である。細胞の移動する先端でアクチン重合が起こり、糸状仮足(フィロポディア)や葉状仮足(ラメリポディア)といった構造を作る。WAVE2を欠損した細胞では顕著に葉状仮足形成が阻害されていたことから、WAVE2は葉状仮足形成に必要不可欠な分子でありArp2/3複合体を制御することで方向性を持った細胞運動全てに重要であることがわかっていた。前年度までの研究では、WAVE2は巨大な複合体を形成することでその局在および安定性を制御されていること、WAVE2複合体は完全に活性化されうることが示唆された。IRSp53はWAVE2複合体に含まれないタンパク質である。IRSp53がRacおよびPIP3依存的に刺激に応じたWAVE2およびWAVE2複合体の活性制御を行いうることを示した。さらにIRSp53のN末部のドメイン(RCBドメイン)のみを細胞に発現させると、細胞の外側にむかって、ラメリポディアやフィロポディアと同じ方向にのびる細胞膜のみから成るように見える突起形成がみられたことから、アクチン重合に先立って細胞膜が細胞の外側に向かって変形することが、ラメリポディアやフィロポディア形成などの突起形成に重要であることが示唆された。本年度の研究はIRSp53およびWAVE2の葉状仮足における局在を詳細に観察し、IRSp53がWAVE2に先立って局在することを示した。また、細胞膜の微細な形態変化を検出するためのFRETprobeの開発に着手した。
また、同様の機構が繊維芽細胞や上皮細胞だけでなく血小板のような血球系細胞でも働いている事を見出した。興味深いことに、血球細胞ではこれらの仮足形成機構が細胞の分化にも関わっている事が示唆された。

Research Products

• [Journal Article] IRSp53: crossing the road of membrane and actin dynamics in the for mation of membrane protrusions. (2008)
  • Author(s): Scita, G., Confalonieri, S., Lappalainen, P. & Suetsugu, S.
  • Journal Title: Trends Cell Biol. 18 Pages: 52-60
  • Peer Reviewed
• [Journal Article] Actin -related protein 3 (Arp3) is mutated in proteinuric BUF/Mna rats. (2008)
  • Author(s): Akiyama, K., Morita, H., Suetsugu, S., Kuraba, S., Numata, Y., Yamamoto, Y., Inui, K., Ideura, T., Wakisaka, N., Nakano, K., Oniki, H., Takenawa, T., Matsuyama, M. & Yoshimura, A.
  • Journal Title: Mamm Genome. 19 Pages: 41-50
  • Peer Reviewed
• [Journal Article] Crystal structure of human DAAM1 formin homology 2 domain. (2007)
  • Author(s): Yamashita, M., Higashi, T., Suetsugu, S., Sato, Y., Ikeda, T., Shirakawa, R., Kita, T., Takenawa, T., Horiuchi, H., Fukai, S. & Nureki, O.
  • Journal Title: Genes Cells. 12 Pages: 1255-65
  • Peer Reviewed
• [Journal Article] The WAVE2/Abi1 complex differentially regulates megakaryocyte development and spreading: implications for platelet biogenesis and spreading machinery. (2007)
  • Author(s): Eto, K., Nishikii, H., Ogaeri, T., Suetsugu, S., Kamiya, A., Kobayashi, T., Yamazaki, D., Oda, A., Takenawa, T. & Nakau chi, H.
  • Journal Title: Blood 110 Pages: 3637-47
  • Peer Reviewed
• [Journal Article] Male-specific sterility caused by the loss of CR16. (2007)
  • Author(s): Suetsugu, S., Banzai, Y., Kato, M., Fukami, K., Kataoka, Y., Takai, Y., Yoshida, N. & Takenawa, T.
  • Journal Title: Genes Cells. 12 Pages: 721-33
  • Peer Reviewed
• [Journal Article] Synaptopodin protects against proteinuria by disrupting Cdc42:IRSp53:Men$ signaling complexes in kidney podocytes. (2007)
  • Author(s): Yanagida-Asanuma, E., Asanuma, K., Kim, K., Donnelly, M., Young Choi, H., Hyung Chang, J., Suetsugu, S., Tomino, Y., Takenawa, T., Faul, C. & Mundel, P.
  • Journal Title: Am J Pathol. 171 Pages: 415-27
  • Peer Reviewed
• [Journal Article] HSP90 cross-links branched actin filaments induced by N-WASP and the Arp2/3 complex. (2007)
  • Author(s): Park, S. J., Suetsugu, S., Sagara, H. & Takenawa, T.
  • Journal Title: Genes Cells. 12 Pages: 611-22
  • Peer Reviewed
• [Journal Article] Curved EFC/F-BAR-domain dimers are joined end to end into a filament for membrane invagination in endocytosis. (2007)
  • Author(s): Shimada, A. *, Niwa, H. *, Tsujita, K. *, Suetsugu, S. *, Nitta, K., Hanawa-Suetsugu, K., Akasaka, R., Nishino, Y., Toyama, M., Chen, L., Liu, Z.J., Wang, B.C., Yamamoto, M., Terada, T., Miyazawa, A., Tanaka, A., Sugano, S., Shirouzu, M., Nagayama, K., Takenawa, T. & Yokoyama, S.
  • Journal Title: Cell. 129 Pages: 761-72
  • Peer Reviewed
• [Journal Article] Effect of WAVE2 Phosphorylation on Activation of the Arp2/3 Complex. (2007)
  • Author(s): Nakanishi O, Suetsugu S, Yamazaki D, Takenawa T.
  • Journal Title: J Biochem (Tokyo). 141・3 Pages: 319-325
  • NAID: 10020100487
• [Journal Article] Stress-associated MAP kinase fills in the map of filamin-mediated neuronal migration. (2007)
  • Author(s): Suetsugu S, Takenawa T.
  • Journal Title: Dev Cell 12・1 Pages: 3-4
• [Journal Article] The WASP-WAVE protein network : connecting the membrane to the cytoskeleton. (2007)
  • Author(s): Takenawa T, Suetsugu S.
  • Journal Title: Nat Rev Mol Cell Biol. 8・1 Pages: 37-48
• [Journal Article] The RAC binding domain/IRSp53-MIM homology domain of IRSp53 induces RAC-dependent membrane deformation. (2006)
  • Author(s): Suetsugu S, Murayama K, Sakamoto A, Hanawa-Suetsugu K, Seto A, Oikawa T, Mishima C, Shirouzu M, Takenawa T, Yokoyama S.
  • Journal Title: J Biol Chem. 281・46 Pages: 35347-35358
• [Journal Article] Optimization of WAVE2 complex-induced actin polymerization by membrane-bound IRSp53, PIP(3), and Rac. (2006)
  • Author(s): Suetsugu S, Kurisu S, Oikawa T, Yamazaki D, Oda A, Takenawa T.
  • Journal Title: J Cell Biol. 173・4 Pages: 571-585
• [Journal Article] Optimization of WAVE2-complex-induced actin polymerization by membrane-bound IRSp53, PIP3, and Rac. (2006)
  • Author(s): Suetsugu S, Kurisu S, Yamazaki D, Oikawa T, Oda A, Takenawa T.
  • Journal Title: J Cell Biol. (in press)
• [Journal Article] Protein kinase A-dependent increase in WAVE2 expression induced by the focal adhesion protein vinexin. (2006)
  • Author(s): Mitsushima M, Sezaki T, Akahane R, Ueda K, Suetsugu S, Takenawa T, Kioka N.
  • Journal Title: Genes Cells. 11・3 Pages: 281-292
• [Journal Article] Coordination between the actin cytoskeleton and membrane deformation by a novel membrane tubulation domain of PCH proteins is involved in endocytosis. (2006)
  • Author(s): Tsujita K, Suetsugu S, Sasaki N, Furutani M, Oikawa T, Takenawa T.
  • Journal Title: J Cell Biol 172・2 Pages: 269-279
• [Journal Article] WAVE3 functions as a negative regulator of LDOC1 (2005)
  • Author(s): Mizutani K, Koike D, Suetsugu S, Takenawa T.
  • Journal Title: J Biochem(Tokyo) 138・5 Pages: 639-646
  • NAID: 10017348480
• [Journal Article] A novel function of WAVE in lamellipodia : WAVE1 is required for stabilization of lamellipodial protrusions during cell spreading. (2005)
  • Author(s): Yamazaki D, Fujiwara T, Suetsugu S, Takenawa T.
  • Journal Title: Genes Cells. 10・5 Pages: 381-392
• [Journal Article] Interaction of HSP90 to N-WASP leads to activation and protection from proteasome-dependent degradation. (2005)
  • Author(s): Park SJ, Suetsugu S, Takenawa T.
  • Journal Title: EMBO J. 24・8 Pages: 1557-1570
• [Presentation] The rearrangements of actin cytoskeleton dependent on the shapes of the plasma membrane. (2007)
  • Author(s): 末次 志郎
  • Organizer: 第30回日本分子生物学会年会・第80回日本生化学会大会合同大会
  • Place of Presentation: インターコンチネンタルホテル
  • Year and Date: 2007-12-12
• [Presentation] Control of Cell Shape and Motility: new approaches to understand itsmechanistic basis (2007)
  • Author(s): 末次 志郎
  • Organizer: 第40回日本発生生物学会・第59回日本細胞生物学会合同大会
  • Place of Presentation: 福岡国際会議場
  • Year and Date: 2007-05-30
• [Presentation] Connecting the membrane to the actin cytoskeleton by BAR, EFC, and RCB domain proteins and WASP/WAVE proteins (2007)
  • Author(s): 末次 志郎
  • Organizer: Workshop on Cell Migration
  • Place of Presentation: IFOM-IEO Campus in Milan
  • Year and Date: 2007-05-12