細胞周期の制御異常

• Project/Area Number: 12215020
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 秋山 徹 東京大学, 分子細胞生物学研究所, 教授 (70150745)
• Co-Investigator(Kenkyū-buntansha): 畑 裕 東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (80313237)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥309,100,000 (Direct Cost: ¥309,100,000); Fiscal Year 2004: ¥63,000,000 (Direct Cost: ¥63,000,000); Fiscal Year 2003: ¥63,000,000 (Direct Cost: ¥63,000,000); Fiscal Year 2002: ¥63,100,000 (Direct Cost: ¥63,100,000); Fiscal Year 2001: ¥65,000,000 (Direct Cost: ¥65,000,000); Fiscal Year 2000: ¥55,000,000 (Direct Cost: ¥55,000,000)
• Keywords: 細胞周期 / 癌抑制遺伝子 / APC / Asef / β-catenin / Wnt / 裏打ち蛋白 / 接着 / がん抑制遺伝子 / TGF-b / がん細胞 / 細胞運動 / ICAT / KAP3 / BAOP1 / MAG1 / PAPIN / 癌 / Wntシグナル / 細胞接着 / 細胞膜裏打ち蛋白質

Research Abstract

がん細胞における細胞周期制御異常の機構を明らかにすることを目的として研究を進め以下の事実を明らかにした。
1.APCによって活性化されるGEF分子Asefのファミリー分子を見出してAsef2と命名し、細胞の形態、接着、運動の制御に関与することを明らかにした。
2.AsefがJNKシグナルの活性化を引き起こすこと、APCがこの活性を促進することを見出した。
3.AsefおよびAsef2遺伝子欠損マウスを作製した。
4.β-catenin-cadherin複合体に結合しCdc42およびRacを標的とするGAP分子RICSの細胞内局在を明らかにし、結合分子2種類を同定した。
5.β-catenin-TCF複合体による転写活性化を促進し、細胞transformationを促進する作用をもつB9Lの遺伝子欠損マウスを作製し、血管、心臓に異常を起こし胎生致死となることを見出した。
6.β-catenin-TCFの標的遺伝子を検索し、機能未知の膜蛋白質およびzinc finger蛋白質が標的であることを見出した。
7.in silico promoter analysisによりTCFとcross talkしている可能性のあるいくつかの転写因子を同定した。
8.DLGと結合しているRap1特異的なGAP分子SPALのファミリー分子2種類を見出しその性質を明らかにした。
7.タイトジャンクションの裏打ち蛋白質MAGI-1に結合する候補分子Rassf1に類似する分子Rassf6の解析を行った。
8.接着分子JAM4とE3ユビキチンリガーゼLNX1の相互作用の解析を行った。
9.MAGI-1と遺伝性ネフローゼの原因遺伝子産物のひとつであるnephrinの相互作用の解析を行った。

Research Products

• [Journal Article] Identification of BMP and Activin Membrane-bound Inhibitor (BAMBI) (2004)
  • Author(s): T.Sekiya et al.
  • Journal Title: J.Biol.Chem. 279 Pages: 6840-6846
• [Journal Article] Anteriorization of neural fate by ICAT, a negative regulator of Wnt signaling. (2004)
  • Author(s): K.Satoh et al.
  • Journal Title: Proc.Natl.Acad.Sci.USA 101 Pages: 8017-8021
• [Journal Article] DKK1, a negative regulator of Wnt signaling, is a target of the β-catenin /TCF pathway. (2004)
  • Author(s): A.Niida et al.
  • Journal Title: Oncogene 23 Pages: 8520-8526
• [Journal Article] Role of a BCL9-related β-catenin-binding protein, B9L, in tumorigenesis induced by aberrant activation of Wnt signaling. (2004)
  • Author(s): S.Adachi et al.
  • Journal Title: Cancer Res. 64 Pages: 8496-8501
• [Journal Article] DREG, a developmentally regulated G protein-coupled receptor containing two conserved proteolytic cleavage sites. (2004)
  • Author(s): T.Moriguchi et al.
  • Journal Title: Genes Cells 9 Pages: 549-560
• [Journal Article] JAM4 enhances hepatocyte growth factor-mediated branching and scattering of Madin-Darby canine kidney cells. (2004)
  • Author(s): H.Mori et al.
  • Journal Title: Genes Cells 9 Pages: 811-819
• [Publications] Y.Kawasaki: "Mutated APC and Asef are Involved in Migration of Colorectal Tumor Cells"Nature Cell Biology. 5・3. 211-215 (2003)
• [Publications] T.Okabe: "RICS, a novel GTPase-Activating Protein for Cdc42 and Rac1, is involved in the β-catenin-N-cadherin and NMDA receptor signaling.β"J.Biol.Chem.. 278. 9920-9927 (2003)
• [Publications] S.Nada: "Identification of PSD-93 as a substrate for the Src family tyrosine kinase Fyn"J.Biol.Chem.. 278. 47610-47621 (2003)
• [Publications] H.Seo: "Incorporation of DUF/FACT into chromatin enhances the accessibility of nucleosomal DNA"Biochem.Biophys.Res.Commun.. 303. 8-13 (2003)
• [Publications] T.Sekiya: "Identification of BMP and Activin Membrane-bound Inhibitor (BAMBI), an Inhibitor of Transforming Growth β-Catenin Pathway in Colorectal Tumorβ Signaling"J Biol Chem.. 279. 6840-6846 (2004)
• [Publications] N.Kakinuma: "Candidate Regions of Tumor Supressor Locus on Chromosome 9q31.1 in Gastric Cancer"Int.J.Cancer.. 109. 71-75 (2004)
• [Publications] A.Hirata: "PDZ domain-binding motif of human T-cell leukemia virus type 1 Tax oncoprotein augments the transforming activity in a rat fibroblast cell line"Virology. 318. 327-336 (2004)
• [Publications] K.Haraguchi: "Activation of beta-catenin-TCF-mediated transcription by non-receptor tyrosine kinase v-Src"Biochem Biophys Res Commun.. 313. 841-844 (2004)
• [Publications] T.Moriguchi: "DREG, a developmentally regulated G protein-coupled receptor containing two conserved proteolytic cleavage sites"Gnes Cells. (in press). (2004)
• [Publications] Y.Hirabayashi: "The Wnt-β-catenin pathway directs neuronal differentiation of cortical neural precursor cells"Development. (in press). (2004)
• [Publications] N.Kakinuma: "Improved In-Gel Competitive Reassociation technique can detect Epstein-Barr virus genome in gastric cancer"J.Electrophoresis. (in press). (2004)
• [Publications] T.Okabe: "RICS, a novel GTPase-Activating Protein for Cdc42 and Rac1, is involved in the b-catenin-N-cadherin and NMDA receptor signaling"J. Biol. Chem.. (in press).
• [Publications] Y.Kawasaki: "Mutated APC and Asef are Involved in Migration of Colorectal Tumor Cells"Nature Cell Biology. 5(3). 211-215 (2003)
• [Publications] T.Jimbo: "Identification of link between the Tumor Suppressor APC and the Kinesin Superfamily of Proteins"Nature Cell Biol.. 4. 323-327 (2002)
• [Publications] T.Sekiya: "Overexpression of Icat Induces G2 Arrest and Cell Death in Tumor Cells Mutant for APC, β-catenin or Axin"Cancer Res.. 62. 3322-3326 (2002)
• [Publications] B.C.Roy: "SPAL, a Rap-specific GTPase Activating Protein, is Present in the NMDA receptor-PSD-95 Complex in the Hippocampus"Genes Cells. 7. 607-617 (2002)
• [Publications] K.Kohu: "The SH3, HOOK and Guanylate Kinase-like Domains of hDLG are Important for its Cytoplasmic Localization"Genes Cells. 7. 707-715 (2002)
• [Publications] Kim M.: "Binding of the mammalian homolog of he Drosophila discs large tumor suppressor protein to the ribosome receptor"Biochem Biophys Res Commun.. 294. 1151-1154 (2002)
• [Publications] T.Koyama: "Mutation and expression of the β-catenin-interacting rotein ICAT in human colorectal tumors"Jpn. J. Clin. Oncol.. 358-362 (2002)
• [Publications] F.Ogawa: "Binding of the Human Homolog of the Drosophila Discs Large Tumor Suppressor Protein to the Mitochondrial Ribosomal Protein MRP-S34"Biochem. Biophys. Res. Commun.. 294. 1151-1154 (2002)
• [Publications] Takemura M.: "Phosphorylation-dependent migration of retinoblastoma protein into the nucleolus triggered by binding to nucleophosmin/B23"Exp. Cell Res.. 276. 233-241 (2002)
• [Publications] H.Seo: "Incorporation of DUF/FACT into chromatin enhances the accessibility of nucleosomal DNA"Biochem. Biophys. Res. Commun.. (in press). (2002)
• [Publications] Ohno H.: "Localization of p0071-interacting proteins, plakophilin-related armadillo-repeat protein-interacting protein and ERBIN, in epithelial cells"Oncogene. 21. 7042-7049 (2002)
• [Publications] Yao I.: "Synaptic localization of SAPAP1, a synaptic membrane-associated protein"Genes Cells. (in press).
• [Publications] Hirabayashi S.: "C2PA is a nuclear protein implicated in heat shock response"J. Cell. Biochem. 87. 65-74 (2002)
• [Publications] Yao I.: "Synaptic and nuclear localization of brain-enriched guanylate kinase-associated protein"J. Neurosci.. 22. 5354-5364 (2002)
• [Publications] Iida J.: "Synaptic localization of membrane-associated guanylate kinase-interacting protein mediated by the pleckstrin homology domain"Eur. J. Neurosci.. 15. 1493-1498 (2002)
• [Publications] T.jimbo: "Link between the Tumor Suppressor APC and the Kinesin Superfamily of Proteins"Nature Cell Biology. (In press). (2002)
• [Publications] K.Kohu: "The SH3, HOOK and GuanylateKinase-like Domains of hDLG are Important for its Cytoplasmic Localization"Genes Cells. (In press). (2002)
• [Publications] T.Sekiya: "Overexpression of ICAT Induces G2 Arrest and Cell Death in Tumor CellsMutant for APC, β-catenin or Axin"Cancer Research. (In press). (2002)
• [Publications] B.C.Roy: "SPAL, a Rap-specific GTPase Activating Protein, is Present in the NMDA receptor-PSD-95 Complex in the Hippocampus"Genes Cells. (In press). (2002)
• [Publications] M.Takemura: "Stimulation of DNA polymerase aactivity by Cdk2-phosphorylated Rb protein. Biochem. Biophys."Res. Commun.. 282. 984-990 (2001)
• [Publications] W.Nishimura: "Interaction of synaptic scaffolding molecule (S-SCAM) and β-catenin"J. Neurosci.. 22-3. 757-765 (2002)
• [Publications] J.Iida: "Synaptic localization of membrane-associated guanylate kinase-interacting protein mediated by the pleckstrin homology domain"Eur. J. Neurosci.. (In press). (2002)
• [Publications] T.Ishidate: "The APC-hDLG Complex Negatively Regulates Cell Cycle Progression from the G0/G1 to SPhase."Oncogene. 19. 365-372 (2000)
• [Publications] H.Nakagawa: "EB3, a novel member of the EB1 family preferentially expressed in the central nervous system, bins to a CNS-specific."Oncogene. 19. 210-216 (2000)
• [Publications] H.Nakagawa: "APCL, a central nervous system-specific homolog of adenomatous polysois coli tumor suppressor, binds to p53-binding protein 2 and translocates."Cancer Res.. 60. 101-105 (2000)
• [Publications] K.Kawahara.: "Downregulation of b-catenin by the Colorectal Tumor Suppressor APC Requires Association with Axin and β-catenin."J.Biol.Chem. 275. 8369-8374 (2000)
• [Publications] T.Akiyama: "Wht/β-catenin signaling."Cytokine & Growth Factor Reviews. 11. 273-282 (2000)
• [Publications] H.Yanai: "The Colorectal Tumor Suppressor APC is Present in the NMDA-receptor-PDS-95 Complex in the Brain."Genes Cells. 5. 815-822 (2000)
• [Publications] Y.Kawasaki: "Asef, a Link between the Tumor Suppressor APC and G-protain Signaling."Science. 289. 1194-1197 (2000)
• [Publications] H.Haraguchi: "The hDLG-associated protein DAP interacts with dynein light chain and neuronal nitric oxide synthase."Gene Cells. 5. 905-911 (2000)
• [Publications] Y.Dabashi: "APC protein is required for initiation of neuronal differentiation in rat pheochromocytoma PC12 cells."Biochem.Biopys.Res.Commun.. 279. 685-691 (2000)
• [Publications] T.Senda: "Expression of APC in the mouse kideney and its colocalization with A-canenin and hDLG."J.Histochem.Cytochem.. (in press).
• [Publications] Kohu,K.: "Localization of the Novel GAP Family Protein SPAL in Epithelial Cells."Acta Histochem.Cytochem.. (in press).
• [Publications] Kakinuma,N: "APC is Colocalized with β-catenin and hDLG in the Henle's Loop of the Mouse Kidney."Acta Histochem.Cytochem.. (in press).
• [Publications] Tago,K.: "Inhibition of Wntsignaling by ICAT, a Novel β-catenin-interacting Protein."Gense Dev.. 14. 1741-1749 (2000)
• [Publications] Nishimura,W: "Localization of BAP1/MAGI1 at adherens junctions in NRK cell : the polarized targeting mediated by the carboxyl-teminal PDZ domains."J.Cell.Physiol.. 185. 358-365 (2000)
• [Publications] Deguchi,M: "PAPIN : a novel multiple PDZ protein interacting with neural plakophilin-related armadillo-repeat protein/β-catenin and P0071"J.Biol.Chem.. 275. 29875-29880 (2000)
• [Publications] yao,I.: "Association of membrane-associated guanylate kinase-interacting protein-1 with Raf-1."Biochem.Biophys.Res.Commun. 270. 538-542 (2000)