2002 年度実績報告書

細胞増殖のシグナリング機構

研究課題

研究課題/領域番号	12215079
研究機関	大阪大学
研究代表者	高井義美大阪大学, 医学系研究科, 教授 (60093514)
研究分担者	清水一也大阪大学, 医学系研究科, 助手 (50335353) 入江賢児大阪大学, 医学系研究科, 助教授 (90232628) 中西宏之大阪大学, 医学系研究科, 助教授 (80314318)
キーワード	細胞増殖 / 細胞接着 / 細胞運動 / Nectin-Afadin系 / Cadherin-Catenin系 / Cdc42 / Rac / がん
研究概要	細胞増殖のシグナル系と細胞の接着・運動のシグナル系との間には密接なクロストークが存在し、細胞の接着阻害の破綻が、がん細胞の増殖、浸潤、転移へとつながる。私共は、新しい細胞間接着機構Nectin-Afadin系を見出し、アドヘレンスジャンクション(AJ)に局在することを明らかにしている。Nectin-Afadin系は、Cadherin-Catenin系と協調してAJの形成を制御している。また、Rhoファミリー低分子量G蛋白質のCdc42とRacがNectin-Afadin系とCadherin-Catenin系を介して細胞間接着を制御していることを明らかにしている。本年度は、Nectin-Afadin系、Cdc42/Racを介した細胞の接着と運動のシグナル系を詳細に解析し、以下の成果を得た。 (1)細胞間接着形成におけるNectin-Afadin系の機能と作用機構:Nectinのトランス結合を阻害するNectinインヒビターおよびNectinの細胞外領域を結合したビーズを用いて、Nectin-Afadin系がAJ形成だけでなく、タイトジャンクション形成においても重要な役割を果たしていることを明らかにした。上皮細胞に肝細胞増殖因子やホルボールエステルを作用すると細胞間接着が破壊され、運動能が亢進して遊走するが、このときNectinの細胞外領域がある種のメタロプロテアーゼによって切断されることを見出した。また、Nectinの発現および切断は、ある種のがん細胞で亢進していた。以上の結果より、Nectin-Afadin系による細胞間接着の制御機構が明らかとなり、がん細胞における細胞間接着能の低下との関連が示唆された。 (2)Nectin-Afadin系による低分子量Gタンパク質の活性制御機構と極性形成:Nectinのトランス結合がRhoファミリーの低分子量Gタンパク質であるCdc42とRacの両者を活性化することを明らかにした。さらに、ネクチンのトランス結合によって活性化されたCdc42とRacはc-Jun NH_2-term in al kinase(JNK)を活性化することを明らかにした。Nectinが極性タンパク質Par3に直接結合し、その局在に関与することを明らかにした。以上の結果から、Nectin-Afadin系が、直接および間接的に細胞間接着形成、上皮細胞の極性形成を制御していると考えられた。このように、本年度の研究は予想以上に進展し、当初の目的を達成することができた。

研究成果
(27件)

すべてその他

すべて文献書誌 (27件)

[文献書誌] Nishimura, M.: "JEAP, a novel component of tight junctions in exocrine cells"J.Biol.Chem.. 277・7. 5583-5587 (2002)
[文献書誌] Nagano, F.: "Rabconnectin-3, a novel protein that binds both GDP/GTP exchange protein and GTPase-activating protein for Rab3 small G protein family"J.Biol.Chem.. 277・12. 9629-9632 (2002)
[文献書誌] Pokutta, S.: "Biochemical and structural definition of the I-afadin-and actin-binding sites of α-catenin"J.Biol.Chem.. 277・21. 18868-18874 (2002)
[文献書誌] Kawakatsu, T.: "Trans-interactions of nectins induce formation of filopodia and lamellipodia through the respective activation of Cdc42 and Rac small G proteins"J.Biol.Chem.. 277・52. 50749-50755 (2002)
[文献書誌] Mizoguchi, A.: "Nectin : An adhesion molecule involved in formation of synapses"J.Cell Biol.. 156・3. 555-565 (2002)
[文献書誌] Ohtsuka, T.: "CAST : A novel protein of the cytomatrix at the active zone of synapses that forms a ternary complex with RIM1 and Munc13-1"J.Cell Biol.. 158・3. 577-590 (2002)
[文献書誌] Yamaguchi, K.: "A GDP/GTP exchange protein for the Rab3 small G protein family up-regulates post-docking step of synaptic exocytosis in the central synapse"Proc.Natl.Acad.Sci.. 99・22. 14536-14541 (2002)
[文献書誌] Ozaki-Kuroda, K.: "Nectin couples cell-cell adhesion and the actin scaffold at heterotypic testicular junctions"Curr.Biol.. 12・13. 1145-1150 (2002)
[文献書誌] Yamamoto, Y.: "Localization of mLin-7 at nectin-based cell-cell junctions"Oncogene. 21・16. 2545-2554 (2002)
[文献書誌] Peng, Y.F.: "Restoration of E-cadherin-based cell-cell adhesion by overexpression of netin in HSC-39 cells, a human signet ring cell gastric cancer cell line"Oncogene. 21・26. 4108-4119 (2002)
[文献書誌] Fukuhara, A.: "Involvement of nectin in the localization of junctional adhesion molecule at tight junctions"Oncogene. 21・50. 7642-7655 (2002)
[文献書誌] Momose, Y.: "Role of the second immunoglobulin-like loop of nectin in cell-cell adhesion"Biochem.Biophys.Res.Commun.. 293・1. 45-49 (2002)
[文献書誌] Tanaka, Y.: "Ectodomain shedding of nectin-1α by SF/HGF and TPA in MDCK cells"Biochem.Biophys.Res.Commun.. 299・3. 472-478 (2002)
[文献書誌] Satoh, K.: "Identification of activity-regulated proteins in the postsynaptic density fraction"Genes Cells. 7・2. 187-197 (2002)
[文献書誌] Kim, Y.: "Association of frabin with specific actin and membrane structures"Genes Cells. 7・4. 413-420 (2002)
[文献書誌] Fukuhara, A.: "Role of nectin in organization of tight junctions in epithelial cells"Genes Cells. 7・10. 1059-1072 (2002)
[文献書誌] Ohtakara, K.: "Densin-180, a synaptic protein, links to PSD-95 through its direct interaction with MAGUIN-1"Genes Cells. 7・11. 1149-1160 (2002)
[文献書誌] Manabe, N.: "Association of ASIP/mPAR-3 with adherens junctins of mouse neuroepithelial cells"Dev.Dyn.. 225・1. 61-69 (2002)
[文献書誌] Asada, M.: "ADIP, a novel afadin and α-actinin-binding protein localized at cell-cell adherens junctions"J.Biol.Chem.. 278・6. 4103-4111 (2003)
[文献書誌] Takekuni, K.: "Direct binding of cell polarity protein PAR-3 to cell-cell adhesion molecule nectin at neuroepithelial cells of developing mouse"J.Biol.Chem.. 278・8. 5497-5500 (2003)
[文献書誌] Yasumi, M.: "Role of each immunoglobulin-like loop of nectin for its cell-cell adhesion activity"Biochem.Biophys.Res.Commun.. 302・1. 61-66 (2003)
[文献書誌] Honda, T.: "Antagonistic and agonistic effects of an extracellular fragment of nectin on formation of E-cadherin-based cell-cell adhesion"Genes Cells. 8・1. 51-63 (2003)
[文献書誌] Tanaka, Y.: "Role of nectin in formation of E-cadherin-based adherens junctions in keratinocytes -Analysis with the N-cadherin dominant negative mutant"Mol.Biol.Cell. (in press). (2003)
[文献書誌] Toi, H.: "She4p/Dim1p interacts with the motor domain of unconventional myosins in the budding yeast, Saccharomyces cerevisiae"Mol.Biol.Cell. (in press). (2003)
[文献書誌] Katata, T.: "Involvement of nectin in the localization of IQGAP1 at the cell-cell adhesion sites through the actin cytoskeleton in Madin-Darby canine kidney cells"Oncogene. (in press). (2003)
[文献書誌] Honda, T.: "Cdc42 and Rac small G proteins activated by trans-interactions of nectins are involved in activation of c-Jun N-terminal kinase, but not in association of nectins and E-cadherin to form adherens junctions, in fibroblasts"Genes Cells. (in press). (2003)
[文献書誌] Inagaki, M.: "Nectin-dependent localization of ZO-1 at puncta adhaerentia junctions between the mossy fiber terminals and the dendrites of the pyramidal cells in the CA3 area of adult mouse hippocampus"J.Comp.Neurol.. (in press). (2003)

2002 年度 実績報告書

細胞増殖のシグナリング機構

研究代表者

高井 義美 大阪大学, 医学系研究科, 教授 (60093514)

研究成果

[文献書誌] Nishimura, M.: "JEAP, a novel component of tight junctions in exocrine cells"J.Biol.Chem.. 277・7. 5583-5587 (2002)

[文献書誌] Nagano, F.: "Rabconnectin-3, a novel protein that binds both GDP/GTP exchange protein and GTPase-activating protein for Rab3 small G protein family"J.Biol.Chem.. 277・12. 9629-9632 (2002)

[文献書誌] Pokutta, S.: "Biochemical and structural definition of the I-afadin-and actin-binding sites of α-catenin"J.Biol.Chem.. 277・21. 18868-18874 (2002)

[文献書誌] Kawakatsu, T.: "Trans-interactions of nectins induce formation of filopodia and lamellipodia through the respective activation of Cdc42 and Rac small G proteins"J.Biol.Chem.. 277・52. 50749-50755 (2002)

[文献書誌] Mizoguchi, A.: "Nectin : An adhesion molecule involved in formation of synapses"J.Cell Biol.. 156・3. 555-565 (2002)

[文献書誌] Ohtsuka, T.: "CAST : A novel protein of the cytomatrix at the active zone of synapses that forms a ternary complex with RIM1 and Munc13-1"J.Cell Biol.. 158・3. 577-590 (2002)

[文献書誌] Yamaguchi, K.: "A GDP/GTP exchange protein for the Rab3 small G protein family up-regulates post-docking step of synaptic exocytosis in the central synapse"Proc.Natl.Acad.Sci.. 99・22. 14536-14541 (2002)

[文献書誌] Ozaki-Kuroda, K.: "Nectin couples cell-cell adhesion and the actin scaffold at heterotypic testicular junctions"Curr.Biol.. 12・13. 1145-1150 (2002)

[文献書誌] Yamamoto, Y.: "Localization of mLin-7 at nectin-based cell-cell junctions"Oncogene. 21・16. 2545-2554 (2002)

[文献書誌] Peng, Y.F.: "Restoration of E-cadherin-based cell-cell adhesion by overexpression of netin in HSC-39 cells, a human signet ring cell gastric cancer cell line"Oncogene. 21・26. 4108-4119 (2002)

[文献書誌] Fukuhara, A.: "Involvement of nectin in the localization of junctional adhesion molecule at tight junctions"Oncogene. 21・50. 7642-7655 (2002)

[文献書誌] Momose, Y.: "Role of the second immunoglobulin-like loop of nectin in cell-cell adhesion"Biochem.Biophys.Res.Commun.. 293・1. 45-49 (2002)

[文献書誌] Tanaka, Y.: "Ectodomain shedding of nectin-1α by SF/HGF and TPA in MDCK cells"Biochem.Biophys.Res.Commun.. 299・3. 472-478 (2002)

[文献書誌] Satoh, K.: "Identification of activity-regulated proteins in the postsynaptic density fraction"Genes Cells. 7・2. 187-197 (2002)

[文献書誌] Kim, Y.: "Association of frabin with specific actin and membrane structures"Genes Cells. 7・4. 413-420 (2002)

[文献書誌] Fukuhara, A.: "Role of nectin in organization of tight junctions in epithelial cells"Genes Cells. 7・10. 1059-1072 (2002)

[文献書誌] Ohtakara, K.: "Densin-180, a synaptic protein, links to PSD-95 through its direct interaction with MAGUIN-1"Genes Cells. 7・11. 1149-1160 (2002)

[文献書誌] Manabe, N.: "Association of ASIP/mPAR-3 with adherens junctins of mouse neuroepithelial cells"Dev.Dyn.. 225・1. 61-69 (2002)

[文献書誌] Asada, M.: "ADIP, a novel afadin and α-actinin-binding protein localized at cell-cell adherens junctions"J.Biol.Chem.. 278・6. 4103-4111 (2003)

[文献書誌] Takekuni, K.: "Direct binding of cell polarity protein PAR-3 to cell-cell adhesion molecule nectin at neuroepithelial cells of developing mouse"J.Biol.Chem.. 278・8. 5497-5500 (2003)

[文献書誌] Yasumi, M.: "Role of each immunoglobulin-like loop of nectin for its cell-cell adhesion activity"Biochem.Biophys.Res.Commun.. 302・1. 61-66 (2003)

[文献書誌] Honda, T.: "Antagonistic and agonistic effects of an extracellular fragment of nectin on formation of E-cadherin-based cell-cell adhesion"Genes Cells. 8・1. 51-63 (2003)

[文献書誌] Tanaka, Y.: "Role of nectin in formation of E-cadherin-based adherens junctions in keratinocytes -Analysis with the N-cadherin dominant negative mutant"Mol.Biol.Cell. (in press). (2003)

[文献書誌] Toi, H.: "She4p/Dim1p interacts with the motor domain of unconventional myosins in the budding yeast, Saccharomyces cerevisiae"Mol.Biol.Cell. (in press). (2003)

[文献書誌] Katata, T.: "Involvement of nectin in the localization of IQGAP1 at the cell-cell adhesion sites through the actin cytoskeleton in Madin-Darby canine kidney cells"Oncogene. (in press). (2003)

[文献書誌] Honda, T.: "Cdc42 and Rac small G proteins activated by trans-interactions of nectins are involved in activation of c-Jun N-terminal kinase, but not in association of nectins and E-cadherin to form adherens junctions, in fibroblasts"Genes Cells. (in press). (2003)

[文献書誌] Inagaki, M.: "Nectin-dependent localization of ZO-1 at puncta adhaerentia junctions between the mossy fiber terminals and the dendrites of the pyramidal cells in the CA3 area of adult mouse hippocampus"J.Comp.Neurol.. (in press). (2003)

2002 年度実績報告書

高井義美大阪大学, 医学系研究科, 教授 (60093514)