1999 年度実績報告書

低分子量G蛋白質の活性制御機構と作用機構

研究課題

研究課題/領域番号	10044285
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	的崎尚大阪大学, 医学系研究科, 助手 (80252782) 中西宏之大阪大学, 医学系研究科, 助手 (80314318) 佐々木卓也大阪大学, 医学系研究科, 助教授 (40241278)
キーワード	低分子量G蛋白質 / Rho / Rab / Rabphilin-11 / BNT1 / RabGD1 / SHP-2 / Frabin
研究概要	現在、低分子量G蛋白質ば、種々の重要な細胞機能を制御していることが明らかになっており、その活性化機構と作用機構を分子レベルで解明することが、医学・生物学領域において重要な課題となっている。本年度の本研究において、私共は、RhoとRabファミリー低分子量G蛋白質の活性化機構と作用機構について解析を行い、以下の成果を得た。 1.Rhoファミリーの活性化機構:私共は、チロシンホスファターゼSHP-2がRhoの活性化を制御し、細胞運動の制御に関与していることを明らかにした。また、Cdc42の活性制御因子FrabinとCdc42を介する系が細胞運動の最も上流に位置する可能性を示した。 2.Rhoファミリーの作用機構:私共は、出芽酵母において、Rhoの標的蛋白質であるBNI1を見い出しているが、BNI1がアクチン細胞骨格を介して微小管系細胞骨格をも制御していることを明らかにした。 3.Rabファミリーの活性化機構:私共は、Rab3の活性制御因子RabGDIの2個のアイソフォームのうち、神経特異的に発現するRabGDIαのノックアウトマウスを作製した.このマウスの解析から、RabGDIαは、Rab3を不活性型に維持し、Rab3によるシナプス小胞輸送の過度の促進を抑制して神経の過度の興奮を抑える作用を有することを明らかにした。 4.RhoとRabファミリーの脇調的作用:私共は、RhoとRabファミリーが脇調的に作用し、CadherinやHGF受容体(c-Met)を含む小胞の輸送を制御して細胞の運動や接着を制御していることを明らかにした。また、Rsb11の標的蛋白質であるRabphin-11を見い出し、Rab11-Rabphilin11系が細胞運動に関与することを示した。海外研究分担者との共同研究という形で論文は発表できなかったが、Schuhlman博士やTamanoi博士との研究打ち合わせや彼等の直接の研究技術指導により、本年度の研究は上記のように予想以上に進展し、当初の目的はほぼ達成できた。

研究成果

(24件)

すべてその他

すべて文献書誌 (24件)

[文献書誌] Yao, I.: "MAGUIN - A novel neuronal menbrane-associated guanylate kinase-interacting protein"J. Biol. Chem.. 274・17. 11889-11896 (1999)
[文献書誌] Umikawa, M.: "Association of frabin with the actin cytoskeleton is essential for microspike formation through activation of Cdc42 small G protein"J. Biol. Chem.. 274・36. 25197-25200 (1999)
[文献書誌] Mammoto, A.: "Rab11BP/Rabphilin-11 -A downstream target of rab11 small G protein implicated in vesicle recycling"J. Biol. Chem.. 274・36. 25517-25524 (1999)
[文献書誌] Yao, I .: "Synamon, a novel neuronal protein interacting with synapse-associated protein 90/postsynaptic density-95-associated protein"J. Biol. Chem.. 274・39. 27463-27466 (1999)
[文献書誌] Kawabe, H.: "nArgBP2, a novel neural member of Ponsin/ArgBR2/Vinexin family that interacts with Synapse-associated protein 90/postsynaptic density-95-associated protein (SAPAP)"J. Biol. Chem.. 274・42. 30914-90918 (1999)
[文献書誌] Mandai, K.: "Ponsin/SH3P12 : An 1-afadin- and vinculin-binding protein localized at cell-cell and cell-matrix adherens junctions"J. Cell Biol.. 144・5. 1001-1017 (1999)
[文献書誌] Takahashi. K.: "Nectin/PRR : An immunoglobulin-like cell adhesion molecule recruited to cadherin-based adherens junctions through interaction with afadin, a PDZ domain-containing protein"J. Cell Biol.. 145・3. 539-549 (1999)
[文献書誌] lkeda. W.: "Afadin : a key molecule essential for structural organization of cell-cell junctions of polarized epithelia during embryogenesis"J. Cell Biol.. 146・5. 1117-1132 (1999)
[文献書誌] Nakano. K.: "Distinct actions and cooperative roles of ROCK and mDia in Rho small G protein-induced reorganization of the actin cytoskeleton in MDCK cells"Mol. Biol. Cell. 10・8. 2481-2491 (1999)
[文献書誌] Fujiwara. T.: "Bnilp regulates microtubule-dependent nuclear migration through the actin cytoskeleton in Saccharomyces cerevisiae"Mol. Biol. Cell. 19・12. 8016-8027 (1999)
[文献書誌] Sakisaka. T.: "Different behavior of 1-afadin and neurabin-II during the formation and destruction of cell-cell adherens junction"Oncogene. 18・8. 1609-1618 (1999)
[文献書誌] Irle. M.: "Isolation and characterization of mammalian homologues of Caenorhabditis elegans lin-7 - localization at cell-cell junctions"Oncogene. 18.18. 2811-2817 (1999)
[文献書誌] Kodama, A.: "Involvement of Cdc42 small G protein in cell-cell adhesion. migration, and morphology of MDCK cells"Oncogene. 18・27. 3996-4006 (1999)
[文献書誌] Togawa, A.: "Progressive impairment of kidneys and reproductive organs in mice lacking Rho GDIα"Oncogene. 18・39. 5373-5380 (1999)
[文献書誌] Kamei, T.: "Coendocytosis of cadherin and c-Met coupled to disruption of cell-cell adhesion in MDCK cells - Regulation by Rho, Rac. and Rab small G proteins"Oncogene. 18・48. 6776-6784 (1999)
[文献書誌] Kikyo. M.: "AnFH domein-containing Bnrlp is a multifunctional protein interacting with a variety of cytoskeletal proteins in Saccharomyces cerevisiae"Oncogene. 18・50. 7046-7054 (1999)
[文献書誌] lde, N.: "Localization of membrane-associated guanylate kinase (MAGI) -1/BAI-associated protein (BAP) 1 at tight junctions of epithelial cells"Oncogene. 18・54. 7810-7815 (1999)
[文献書誌] Asakura, T.: "Similar and differential behavior between the nectin-afadin-ponsin and cadherin-catenin systems during the formation and disruption of the polarized junctional alignment in epithelial cells"Genes Cells. 4・10. 573-581 (1999)
[文献書誌] Sakaguchi. G.: "Doc2α is an activity-dependent modulator of excitatory synaptic transmission"Eur. J. Neurosci.. 11・12. 4262-2468 (1999)
[文献書誌] Miyahara, M.: "Interaction of nectin with afadin is necessary for its clustering at cell-cell contact sites but not for its cis dimerization or trans interaction"J. Biol. Chem.. 275・1. 613-618 (2000)
[文献書誌] Satoh-Horikawa, K.: "Nectin-3 : A new member of immunoglobulin-like cell adhesion molecules that shows homophilic and heterophilic cell-cell adhesion activities"J. Biol. Chem.. (in press). (2000)
[文献書誌] Ide, N.: "Distinct localization of the S-SCAM and PSD-95/SAP90 families at cell-cell junctions"Oncogene. (in press). (2000)
[文献書誌] Sakisaka. T.: "Requirement of interaction of Nectin-1/HveC with afadin for efficient entry of herpes simplex virus type 1"J. Virol.. (in press). (2000)
[文献書誌] Mammoto. A.: "Physical and functional interaction of rabphilin-11 with mammalian sec13 protein. Implication in vesicle trafficking"J. Biol. Chem.. (in press). (2000)

1999 年度 実績報告書

低分子量G蛋白質の活性制御機構と作用機構

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] Yao, I.: "MAGUIN - A novel neuronal menbrane-associated guanylate kinase-interacting protein"J. Biol. Chem.. 274・17. 11889-11896 (1999)

[文献書誌] Umikawa, M.: "Association of frabin with the actin cytoskeleton is essential for microspike formation through activation of Cdc42 small G protein"J. Biol. Chem.. 274・36. 25197-25200 (1999)

[文献書誌] Mammoto, A.: "Rab11BP/Rabphilin-11 -A downstream target of rab11 small G protein implicated in vesicle recycling"J. Biol. Chem.. 274・36. 25517-25524 (1999)

[文献書誌] Yao, I .: "Synamon, a novel neuronal protein interacting with synapse-associated protein 90/postsynaptic density-95-associated protein"J. Biol. Chem.. 274・39. 27463-27466 (1999)

[文献書誌] Kawabe, H.: "nArgBP2, a novel neural member of Ponsin/ArgBR2/Vinexin family that interacts with Synapse-associated protein 90/postsynaptic density-95-associated protein (SAPAP)"J. Biol. Chem.. 274・42. 30914-90918 (1999)

[文献書誌] Mandai, K.: "Ponsin/SH3P12 : An 1-afadin- and vinculin-binding protein localized at cell-cell and cell-matrix adherens junctions"J. Cell Biol.. 144・5. 1001-1017 (1999)

[文献書誌] Takahashi. K.: "Nectin/PRR : An immunoglobulin-like cell adhesion molecule recruited to cadherin-based adherens junctions through interaction with afadin, a PDZ domain-containing protein"J. Cell Biol.. 145・3. 539-549 (1999)

[文献書誌] lkeda. W.: "Afadin : a key molecule essential for structural organization of cell-cell junctions of polarized epithelia during embryogenesis"J. Cell Biol.. 146・5. 1117-1132 (1999)

[文献書誌] Nakano. K.: "Distinct actions and cooperative roles of ROCK and mDia in Rho small G protein-induced reorganization of the actin cytoskeleton in MDCK cells"Mol. Biol. Cell. 10・8. 2481-2491 (1999)

[文献書誌] Fujiwara. T.: "Bnilp regulates microtubule-dependent nuclear migration through the actin cytoskeleton in Saccharomyces cerevisiae"Mol. Biol. Cell. 19・12. 8016-8027 (1999)

[文献書誌] Sakisaka. T.: "Different behavior of 1-afadin and neurabin-II during the formation and destruction of cell-cell adherens junction"Oncogene. 18・8. 1609-1618 (1999)

[文献書誌] Irle. M.: "Isolation and characterization of mammalian homologues of Caenorhabditis elegans lin-7 - localization at cell-cell junctions"Oncogene. 18.18. 2811-2817 (1999)

[文献書誌] Kodama, A.: "Involvement of Cdc42 small G protein in cell-cell adhesion. migration, and morphology of MDCK cells"Oncogene. 18・27. 3996-4006 (1999)

[文献書誌] Togawa, A.: "Progressive impairment of kidneys and reproductive organs in mice lacking Rho GDIα"Oncogene. 18・39. 5373-5380 (1999)

[文献書誌] Kamei, T.: "Coendocytosis of cadherin and c-Met coupled to disruption of cell-cell adhesion in MDCK cells - Regulation by Rho, Rac. and Rab small G proteins"Oncogene. 18・48. 6776-6784 (1999)

[文献書誌] Kikyo. M.: "AnFH domein-containing Bnrlp is a multifunctional protein interacting with a variety of cytoskeletal proteins in Saccharomyces cerevisiae"Oncogene. 18・50. 7046-7054 (1999)

[文献書誌] lde, N.: "Localization of membrane-associated guanylate kinase (MAGI) -1/BAI-associated protein (BAP) 1 at tight junctions of epithelial cells"Oncogene. 18・54. 7810-7815 (1999)

[文献書誌] Asakura, T.: "Similar and differential behavior between the nectin-afadin-ponsin and cadherin-catenin systems during the formation and disruption of the polarized junctional alignment in epithelial cells"Genes Cells. 4・10. 573-581 (1999)

[文献書誌] Sakaguchi. G.: "Doc2α is an activity-dependent modulator of excitatory synaptic transmission"Eur. J. Neurosci.. 11・12. 4262-2468 (1999)

[文献書誌] Miyahara, M.: "Interaction of nectin with afadin is necessary for its clustering at cell-cell contact sites but not for its cis dimerization or trans interaction"J. Biol. Chem.. 275・1. 613-618 (2000)

[文献書誌] Satoh-Horikawa, K.: "Nectin-3 : A new member of immunoglobulin-like cell adhesion molecules that shows homophilic and heterophilic cell-cell adhesion activities"J. Biol. Chem.. (in press). (2000)

[文献書誌] Ide, N.: "Distinct localization of the S-SCAM and PSD-95/SAP90 families at cell-cell junctions"Oncogene. (in press). (2000)

[文献書誌] Sakisaka. T.: "Requirement of interaction of Nectin-1/HveC with afadin for efficient entry of herpes simplex virus type 1"J. Virol.. (in press). (2000)

[文献書誌] Mammoto. A.: "Physical and functional interaction of rabphilin-11 with mammalian sec13 protein. Implication in vesicle trafficking"J. Biol. Chem.. (in press). (2000)

1999 年度実績報告書

高井義美大阪大学, 医学系研究科, 教授 (60093514)