イノシトールリン脂質によるWASPファミリー蛋白質の活性抑制

• Project/Area Number: 16209009
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: The University of Tokyo
• Principal Investigator: 竹縄 忠臣 東大, 医科学研究所, 教授 (40101315)
• Co-Investigator(Kenkyū-buntansha): 末次 志郎 東京大学, 医科学研究所, 助手 (70345031) ; 伊集院 壮 東京大学, 医科学研究所, 研究員 (00361626)
• Project Period (FY): 2004 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥46,020,000 (Direct Cost: ¥35,400,000、Indirect Cost: ¥10,620,000); Fiscal Year 2006: ¥14,690,000 (Direct Cost: ¥11,300,000、Indirect Cost: ¥3,390,000); Fiscal Year 2005: ¥14,690,000 (Direct Cost: ¥11,300,000、Indirect Cost: ¥3,390,000); Fiscal Year 2004: ¥16,640,000 (Direct Cost: ¥12,800,000、Indirect Cost: ¥3,840,000)
• Keywords: イノシトールリン脂質 / WAVE2 / 葉状仮足 / PI(3,4,5)P3 / IRSp53 / Rac / 細胞遊走 / PI3-キナーゼ

Research Abstract

イノシトールリン脂質結合の性質を明かにし、WASPファミリー蛋白質による葉状仮足形成および方向性を持った細胞遊走へのイノシトールリン脂質結合の関与を解明することを目的とした。WASPファミリー蛋白質の一つWAVE2は葉状仮足形成にあたり、Racの下流でArp2/3複合体を活性化してアクチンの重合を促進する。WAVE2は大きな複合体として存在し、Abi1,Nap1,Sra1及びHSPC300と結合して存在する。IRSp53はRacとWAVE2の橋渡しをする蛋白質で、そのノックダウンは葉状仮足形成を抑制した。しかしWAVE2の細胞間接着部位への局在には変化なかった。
Arp2/3複合体の活性化をin vitroで検証した。膜画分からのWAVE2はIRSp53依存的にフルに活性化された。一方、細胞質画分からのWAVE2は活性化が弱かった。精製したWAVE2はIRSp53によりRacおよびPI(3,4,5)P3依存的に活性化された。よってIRSp53はRacおよびPI(3,4,5)P3とともにWAVE2の活性化を調節していると考えられた。

Research Products

• [Journal Article] Coordination between actin cytoskeletal reorganization and the membrane deformation by a novel membrane tubulation domain of the PCH protein family involved in endocytosis (2006)
  • Author(s): Tsujita K., Sasaki N., Furutani M., Oikawa T., Suetsugu S., Takenawa T.
  • Journal Title: J.Cell Biol. 172・2 Pages: 269-279
• [Journal Article] Molecular mechanisms of invadopodium formation : the role of the N-WASP-Arp2/3 complex pathway and cofilin (2005)
  • Author(s): Yamaguchi, H., Lorenz, M., Kempiak, S., Sarmiento, C., Coniglio, S., Symons, M., Segall, J., Eddy, R., Miki, H., Takenawa, T., Condeelis, J.
  • Journal Title: J.Cell Biol. 168・3 Pages: 441-452
• [Journal Article] Rac-WAVE2 signaling is involved in the invasive and metastatic phenotypes of murine melanoma cells. (2005)
  • Author(s): Kurisu, S., Suetsugu, S., Yamazaki, D., Yamaguchi, H., Takenawa, T.
  • Journal Title: Oncogene 24・8 Pages: 1309-1319
• [Journal Article] Interaction of HSP90 to N-WASP leads to activation and protection from proteasome-dependent degradation. (2005)
  • Author(s): Park, S.J., Suetsugu, S., Takenawa, T.
  • Journal Title: EMBO J. 24・8 Pages: 1557-1570
• [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] CRMP-2 is involved in kinesin-1-dependent transport of the Sra-1/WAVE1 complex and axon formation (2005)
  • Author(s): Kawano Y., Yoshimura T., Tsuboi D., Kawabata S., Kaneko-Kawano T., Shirataki H., Takenawa T., Kaibuchi K.
  • Journal Title: Mol.Cell.Biol. 25・22 Pages: 9920-9935
• [Journal Article] Phospholipase C-{delta}1 and -{delta}3 Are Essential in the Trophoblast for Placental Development. (2005)
  • Author(s): Nakamura Y., Hamada Y., Fujiwara T., Enomoto H., Hiroe T., Tanaka S., Nose M., Nakahara M., Yoshida N., Takenawa T., Fukami K.
  • Journal Title: Mol.Cell.Biol. 25・24 Pages: 10979-10988
• [Journal Article] PtdIns(3,4,5)P3 binding is necessary for WAVE2-induced formation of lamellipodia. (2004)
  • Author(s): Oikawa, T., Yamaguchi, H., Itoh, T., Kato, M., Ijuin, T., Yamazaki, D., Suetsugu, S., Takenawa, T.
  • Journal Title: Nat.Cell Biol. 6 Pages: 420-426
• [Journal Article] Arp2/3 activity is necessary for efficient formation of E-cadherin adhesive contacts. (2004)
  • Author(s): Verma, S., Shewan, A.M., Scott, J.A., Helwani, F.M., den Elzen, N.R., Miki, H., Takenawa, T., Yap, A.S.
  • Journal Title: J Biol.Chem. 279 Pages: 34062-34070
• [Journal Article] Phospholipase C delta-type consists of three isozymes : bovine PLCdelta2 is a homologue of human/mouse PLCdelta4. (2004)
  • Author(s): Irino, Y., Cho, H., Nakamura, Y., Nakahara, M., Furutani, M., Suh, P.G., Takenawa, T., Fukami, K.
  • Journal Title: Biochem.Biophys.Res.Commun. 23 Pages: 537-543
• [Journal Article] WAVE/Scars in Platelets. (2004)
  • Author(s): Oda, A., Miki, H., Wada, I., Yamaguchi, H., Yamazaki, D., Suetsugu, S., Nakajima, M., Nakayama, A., Okawa, K., Miyazaki, H., Matsuno, K., Ochs, H.D, Machesky, L.M., Fujita, H., Takenawa, T.
  • Journal Title: Blood. 2004 Aug 3 Epub 2004 Jul 27 (In press)
• [Journal Article] Regulation of actin cytoskeleton by mDab1 through N-WASP and ubiquitination of mDab1. (2004)
  • Author(s): Suetsugu S., Tezuka T., Morimura T., Hattori M., Mikoshiba K., Yamamoto T., Takenawa T.
  • Journal Title: Biochem J. 384 Pages: 1-8
• [Journal Article] N-WASP and WAVE2 acting downstream of phosphatidylinositol 3-kinase are required for myogenic cell migration induced by hepatocyte growth factor. (2004)
  • Author(s): Kawamura, K., Takano, K., Suetsugu, S., Kurisu, S., Yamazaki, D., Miki, H., Takenawa, T., Endo T.
  • Journal Title: J Biol Chem. 279 Pages: 54862-54871