スフィンゴ脂質による生体膜ドメイン形成と多機能シグナリング

• Project/Area Number: 12140101
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Hokkaido University
• Principal Investigator: 五十嵐 靖之 北海道大学, 大学院・薬学研究科, 教授 (70091965)
• Co-Investigator(Kenkyū-buntansha): 平林 義雄 理化学研究所, 脳科学総合研究センター, ユニットリーダー (90106435) ; 小堤 保則 京都大学, 生命科学研究所, 教授 (70205425) ; 永井 克孝 三菱化学生命科学研究所, 所長 (80072974) ; 清水 孝雄 東京大学, 医学部, 教授 (80127092) ; 西島 正弘 感染症研究所, 細胞化学部, 部長 (60072956)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥26,400,000 (Direct Cost: ¥26,400,000); Fiscal Year 2004: ¥8,500,000 (Direct Cost: ¥8,500,000); Fiscal Year 2003: ¥5,900,000 (Direct Cost: ¥5,900,000); Fiscal Year 2002: ¥6,000,000 (Direct Cost: ¥6,000,000); Fiscal Year 2001: ¥6,000,000 (Direct Cost: ¥6,000,000)
• Keywords: スフィンゴ脂質 / 糖脂質 / スフィンゴシン1-リン酸 / セラミド / スフィンゴシンキナーゼ / 細胞膜ミクロドメイン / シグナル伝達 / G蛋白質 / アポトーシス / 細胞内脂質輸送 / スフィンゴシン1 リン酸

Research Abstract

この1年間に本領域研究は順調に進展しており、各班独自の研究成果に基づく班員相互の連携による共同研究でもいくつか成果をあげている。これまでに領域全体でこの1年間にNature, JBCなどの国際一流誌73報の原著論文が発表された。更に7月には世界中から40名近くの研究者を招き200名規模のスフィンゴ脂質国際シンポジウムを札幌で開催しこれを成功させた。この1年の特記すべき成果としては、(1)セラミドをERからゴルジ体に運搬する遺伝子CERT(Nature)の基質特異性や運搬機構の解析を引き続き押し進めた。(2)スフィンゴ脂質合成に必須の前駆体であるセリンの合成酵素遺伝子のプルキンエ組織特異的ノックアウトマウスを作ることに成功し、新規糖脂質を発見した(3)ゼブラフィッシュ初期発生系における中性セラミダーゼの役割をノックアウト動物を作製して示した(4)スフィンゴ脂質合成阻害作用を持つ免疫抑制剤であるISP-1の酵母の耐性遺伝子SLI1の作用機構を更に検討した。(5)動物細胞スフィンゴミエリン合成酵素のクローニングに成功した(6)酵母細胞膜の非対称性維持機構の存在,特にスフィンゴ脂質とグリセロリン脂質のクロストーク機構の存在を初めて示した(MBC,2004)(7)哺乳類スフィンゴ脂質代謝のうちで最後まで残されていた3-ケトヒドロキシスフィンゴシン還元酵素(FTV-1)のクローニングに成功した、酵母細胞スフィンゴシンキナーゼのリン酸化機構とその分解における役割を明らかにした。

Research Products

• [Journal Article] CERT mediates intermembrane transfer of various molecular species of ceramides. (2005)
  • Author(s): Kumagai K.
  • Journal Title: J.Biol.Chem. (In press)
• [Journal Article] Targeted disruption of the mouse 3-phosphoglycerate dehydrogenase gene causes severe neurodevelopmental defects and results in embryonic lethality. (2004)
  • Author(s): Hirabayashi Y.
  • Journal Title: J.Biol.Chem. 279 Pages: 3573-3577
• [Journal Article] Molecular cloning and functional analysis of zebrafish neutral ceramidase. (2004)
  • Author(s): Yoshimura Y.
  • Journal Title: J.Biol.Chem. 279 Pages: 44012-44022
• [Journal Article] SLI1 (YGR212W) is a major gene conferring resistance to sphingolipidbiosynthesis inhibitor ISP-1, and encodes an ISP-1 N-acetyltransferase in yeast (2004)
  • Author(s): Momoi M.
  • Journal Title: Biochem J. 381 Pages: 321-329
• [Journal Article] Cross talk between sphingolipids and glycerophospholipids in the establishment of plasma memmbrane asymmetry. (2004)
  • Author(s): Kihara A.
  • Journal Title: Mol.Biol.Cell 15(11) Pages: 4949-4959
• [Journal Article] FVT-1 is a mammalian 3-ketodihydrosphingosine reductase with an active site that faces the cytosolic side of the endoplasmic Reticulum Membrane. (2004)
  • Author(s): Kihara A.
  • Journal Title: J.Biol.Cell 279 Pages: 49243-49250
• [Publications] K.Hanada: "Molecular machinery for non-vesicular trafficking of ceramide"Nature. 246. 803-809 (2003)
• [Publications] Yoshida K.: "Targeted disruption of the mouse 3-phosphoglycerate dehydrogenase gene cause severe neurodevelopmental defects and results in embryonic lethality"J.Biol.Chem.. 279. 3573-3577 (2004)
• [Publications] Nagatsuka, Y.: "Carbohydrate-dependent signaling from the phosphatidyiglucoside-based microdomain induces granulocytic differentiation of HL60 cells"Proc.Natl.Acad.Sci.USA. 100. 7454-7459 (2003)
• [Publications] M.Tani: "O-glycosylation of mucin-like domain retains the neutral ceramidase on the plasma membranes as a type II integral membrane protein"J.Biol.Chem.. 278(12). 10523-10530 (2003)
• [Publications] Momoi M.: "SLI1 (YGR212W) is a major gene conferring resistance to sphingolipid biosynthesis inhibitor ISP-1, and encodes ISP-1 N-acetyltransferase in yeas"Biochem J.. (In press).
• [Publications] S.Uemura: "Csg1p and newly identified Csh1p function in mannosylinositol phosphorylceramide synthesis by interacting with Csg2p"J.Biol.Chem.. 278(46). 45049-45055 (2003)
• [Publications] Hanada K.: "Purification of the serine palmitoyltransferase complex responsible for sphingoid base synthesis by using affinity-peptide chromatography techniques"J. Biol. Chem.. 257. 8409-8415 (2000)
• [Publications] Kakio, A.: "Interactions of amyloid beta-protein with various gangliosides in raft-like membranes : importance of GM1 ganglioside-bound form as an endogenous seed for Alzheimer amyloid"Biochemistry. 41・23. 7385-7390 (2002)
• [Publications] Furusato M.: "Molecular cloning and characterization of sphingolipid ceramide N-deacylase from a marine bacterium, Shewanella alga G8"J. Biol. Chem.. 277. 17300-17307 (2002)
• [Publications] Tani M.: "O-Glycosylation of mucin-like domain retains the neutral ceramidase on the plasma membranes as a type II integral membrane protein"J. Biol. Chem.. (In press).
• [Publications] Iwai K.: "Ceramide enhances H202-induced oxidative damage and apoptosis by caspase-3-dependent catalase depletion"J. Biol. Chem.. (In press).
• [Publications] Kihara A.: "Identification and characterization of a saccharomyces cerevisiae gene, RSB1, involved in sphingoid long-chain base release"J. Biol. Chem.. 277(In press). (2002)
• [Publications] Ogawa C.: "Identification and characterization of a novel human sphingosine 1-phosphate phosphohydrolase, hSPP2"J. Biol. Chem.. 278. 1268-1272 (2003)
• [Publications] Kohno T.: "N-glycans of sphingosine-1-phosphate receptor Edg1 regulate ligand-induced receptor internalization"FASEB J.. 16. 983-992 (2002)
• [Publications] Tagami S.: "Ganglioside GM3 participates in the pathological conditions of insulin resistance"J. Biol. Chem.. 277. 3085-3092 (2002)
• [Publications] Kihara A.: "Transmembrane topology and functions of sphingoid long-chain 1-phosphate phosphatase"Gene to Cells. (In press). (2003)
• [Publications] Kihara A.: "Sphimgosine 1-phosphatelyase, SPL, is involved in differentiation of F9 embryonal carcinoma cells toward promitive endoderm"J. Biol. Chem.. (In press). (2003)
• [Publications] 五十嵐靖之: "マイクロドメイン形成と細胞のシグナリング"共立出版. 551 (2003)
• [Publications] Kazuya Kabayama: "Suppression of Integrin Expression and Tumorigenicity by Sulfation of Lactosvlceramide in 3LL Lewis Lung Carcinoma Cells"The Journal of Biological Chemistry. 276(29). 26777-26783 (2001)
• [Publications] Seiichi Tagami: "Ganglioside GM3 Participates in the Pathological Conditions of Insurin Resistance"The Journal of Biological Chemistry. 277(5). 3085-3092 (2002)
• [Publications] Kimura T.: "Sphingosine 1-phosphate may be a major component of plasma lipoproteins responsible for the cytoprotective actions in human umbilical vein endothelial cells"The Journal of Biological Chemistry. 276(34). 31780-31785 (2001)
• [Publications] Ohmori T.: "G(i)-mediated Cas tyrosine phosphorylation in vascular endothelial cells stimulated with sphingosine 1-phosphate : possible involvement in cell motility enhancement in cooperation with Rho-mediated pathways"The Journal of Biological Chemistry. 276(7). 5274-5280 (2001)
• [Publications] Yasuda S.: "A novel inhibitor of ceramide trafficking from the endoplasmic reticulum to the site of sphingomyelin synthesis"The Journal of Biological Chemistry. 276(47). 43994-44002 (2001)
• [Publications] Takayuki Kohno: "N-glycans of Sphingosine 1-phosphate Receptor, Edg-1, Regulate Ligand Induced Receptor Internalization"FASEB Journal. (in press). (2002)
• [Publications] 五十嵐靖之 他編: "マイクロドメイン形成と細胞のシグナリング:スフィンゴ脂質の新しい生物機能"共立出版株式会社. 258 (2002)
• [Publications] Tomoko Funakoshi: "Reconstitution of ATP-and cytosol-dependent transport of de novo synthesized ceramide to the site of sphingomyelin synthesis in semi-intact cells."J.Biol.Chem.. 275. 29938-29945 (2000)
• [Publications] Tani M.: "Molecular cloning of the full-length cDNA encoding mouse neutral ceramidase-A NOBEL BUT HIGHLY CONSERVED GENE FAMILY OF NEUIRAL/ALKALINE CERAMIDASES."J.Biol.Chem... 275・15. 11229-11234 (2000)
• [Publications] Kondo T.: "Suppression of heat shock protein-70 by ceramide in heat shock-induced HL-60 cell apoptosis."J.Biol.Chem.. 275. 8872-8879 (2000)
• [Publications] Yatomi Y.: "Sphingosine 1-phosphate as a major bioactive lysophospholipid released from platelets and interacting with endothelial cells."Blood. 96. 3431-3438 (2000)
• [Publications] Furuya,S.: "L-Serine and glycine serve as major astroglia-derived trophic factors for cerebellar Purkinje neurons."Proc.Natl.Acad.Sci.USA.. 97. 11528-11533 (2000)
• [Publications] Kanazawa T.: "Inhibition of cytokinesis by a lipid metabolite, psychosine."J Cell.Biol. 149. 943-950 (2000)