2000 年度実績報告書

Rhoファミリーによる細胞骨格と細胞接着の制御機構

研究課題

研究課題/領域番号	11480211
研究機関	名古屋大学
研究代表者	貝淵弘三名古屋大学, 医学部, 教授 (00169377)
研究分担者	天野睦紀奈良先端科学技術大学院大学, バイオサイエンス研究科, 助手 (90304170)
キーワード	低分子量GTPase / Rho-キナーゼ / IQGAP1 / カドヘリン / 細胞運動 / 細胞接着 / リン酸化 / CRMP-2
研究概要	Rhoファミリー低分子量GTPase(Rho,Rac,Cdc42)は、細胞外シグナルの下流で主にアクチン細胞骨格と細胞接着を制御していると考えられている。私共は既に、RacとCdc42の標的蛋白質としてIQGAP1を、Rhoの標的蛋白質としてRho-キナーゼとMBSを同定している。本研究では、細胞骨格及び細胞接着制御機構を明らかにすることを目的とし、Rhoファミリー低分子量GTPase及びその標的蛋白質の機能解析を行った。私共は既に、IQGAP1がβ-カテニンと直接結合し、カドヘリン・カテニン複合体からα-カテニンを解離させることにより細胞間接着を負に制御することを明らかにしている。また、活性型RacとCdc42がIQGAP1の機能を阻害し、細胞間接着を正に制御することも明らかにしている。本研究では、このRac/Cdc42/IQGAP1による細胞間接着の制御システムがhepatocyte growth factor(HGF)により誘導されるcell scattering(細胞分散)時の細胞間接着の破壊に関与しているか否か検討した。その結果、Cdc42、Rac及びIQGAP1はα-カテニンの動態制御を介して、cell scattering時の細胞間接着の破壊に関与していることが明らかとなった。また、RacとCdc42の活性化動態を特異的にモニターするassay系の開発に成功し、細胞間接着の再構築時におけるRacとCdc42の活性動態を明らかにした。一方、私共は既に、Rho-キナーゼの基質としてMBSやMLC、adducin、ERMなどを報告しているが、本研究では、新たにRho-キナーゼの基質としてCRMP-2、MAP2、tau、calponin、EF-1αを同定した。さらに、私共はRho-キナーゼによるCRMP-2のリン酸化がLPAによる神経細胞の成長円錐の退縮に重要な役割を果たしていることを明らかにした。以上、本研究の研究目標はほぼ達成できたと考えられる。

研究成果

(20件)

すべてその他

すべて文献書誌 (20件)

[文献書誌] N.Arimura: "Phosphorylation of collapsin response mediator protein-2 by Rho-kinase. Evidence for two separate signaling pathways for growth cone collapse"J.Biol.Chem.. 275. 23973-23980 (2000)
[文献書誌] N.Nakamura: "Phosphorylation of ERM proteins at filopodia induced by Cdc42"Genes Cells. 5. 571-581 (2000)
[文献書誌] M.Amano: "Regulation and functions of rho-associated kinase"Exp.Cell Res.. 261. 44-51 (2000)
[文献書誌] M.Amano: "Purification and in vitro activity of Rho-associated kinase"Methods Enzymol.. 325. 149-155 (2000)
[文献書誌] T.Kaneko: "Identification of calponin as a novel substrate of Rho-kinase"Biochem. Biophys. Res.Commun.. 273. 110-116 (2000)
[文献書誌] A.Kawajiri: "Identification of a novel beta-catenin-interacting protein"Biochem. Biophys. Res.Commun.. 273. 712-717 (2000)
[文献書誌] T.Izawa: "Elongation factor-lalpha is a novel substrate of rho-associated kinase"Biochem. Biophys. Res.Commun.. 278. 72-78 (2000)
[文献書誌] I.Royal: "Activation of cdc42,rac, PAK, and rho-kinase in response to hepatocyte growth factor differentially regulates epithelial cell colony spreading and dissociation"Mol.Biol.Cell. 11. 1709-1725 (2000)
[文献書誌] N.Inagaki: "Activation of Ca^<2+>/calmodulin-dependent protein kinase II within post-synaptic dendritic spines of cultured hippocampal neurons"J.Biol.Chem.. 275. 27165-27171 (2000)
[文献書誌] M.Sone: "Synaptic development is controlled in the periactive zones of drosophila synapses"Development.. 127. 4157-4168 (2000)
[文献書誌] T.Kandabashi: "Inhibition of myosin phosphatase by upregulated rho-kinase plays a key role for coronary artery spasm in a porcine model with interleukin-1beta"Circulation. 101. 1319-1323 (2000)
[文献書誌] S.Miyamoto: "Changes in E-cadherin associated with cytoplasmic molecules in well and poorly differentiated endometrial cancer"Br.J.Cancer. 83. 1168-1175 (2000)
[文献書誌] K.Ihara: "Crystallization and preliminary crystallographic analysis of the rho-binding domain of bovine rho-kinase"Acta.Crystallogr. D.Biol.Crystallogr.. 56. 1042-1044 (2000)
[文献書誌] P.A.Oude Weernink: "Stimulation of phosphatidylinositol-4-phosphate 5-kinase by Rho-kinase"J.Biol.Chem.. 275. 10168-10174 (2000)
[文献書誌] K.Szaszi: "RhoA and rho kinase regulate the epithelial Na^+/H^+ exchanger NHE3. Role of myosin light chain phosphorylation"J.Biol.Chem.. 275. 28599-28606 (2000)
[文献書誌] R.Hashimoto: "Site-specific phosphorylation of neurofilament-L is mediated by calcium/calmodulin-dependent protein kinase II in the apical dendrites during long-term Potentiation"J.Neurochem.. 75. 373-382 (2000)
[文献書誌] M.Kanai-Azuma: "Co-localization of FAM and AF-6, the mammalian homologues of Drosophila faf and canoe, in mouse eye development"Mech.Dev.. 91. 383-386 (2000)
[文献書誌] T.Hamaguchi: "Phosphorylation of CPI-17, an inhibitor of myosin phosphatase, by protein kinase N"Biochem. Biophys. Res.Commun.. 274. 825-830 (2000)
[文献書誌] Y.Fukata: "Rho-Rho-kinase pathway in smooth muscle contraction and cytoskeletal reorganization of non-muscle cells"TRENDS in Pharm.Sci.. 22. 32-39 (2001)
[文献書誌] M.Fukata: "Involvement of IQGAP1, an Effector of Rac1 and Cdc42 GTPases, in Cell-Cell Dissociation during Cell Scattering"Mol.Cell.Biol.. 21(in press). (2001)

2000 年度 実績報告書

Rhoファミリーによる細胞骨格と細胞接着の制御機構

研究代表者

貝淵 弘三 名古屋大学, 医学部, 教授 (00169377)

研究成果

[文献書誌] N.Arimura: "Phosphorylation of collapsin response mediator protein-2 by Rho-kinase. Evidence for two separate signaling pathways for growth cone collapse"J.Biol.Chem.. 275. 23973-23980 (2000)

[文献書誌] N.Nakamura: "Phosphorylation of ERM proteins at filopodia induced by Cdc42"Genes Cells. 5. 571-581 (2000)

[文献書誌] M.Amano: "Regulation and functions of rho-associated kinase"Exp.Cell Res.. 261. 44-51 (2000)

[文献書誌] M.Amano: "Purification and in vitro activity of Rho-associated kinase"Methods Enzymol.. 325. 149-155 (2000)

[文献書誌] T.Kaneko: "Identification of calponin as a novel substrate of Rho-kinase"Biochem. Biophys. Res.Commun.. 273. 110-116 (2000)

[文献書誌] A.Kawajiri: "Identification of a novel beta-catenin-interacting protein"Biochem. Biophys. Res.Commun.. 273. 712-717 (2000)

[文献書誌] T.Izawa: "Elongation factor-lalpha is a novel substrate of rho-associated kinase"Biochem. Biophys. Res.Commun.. 278. 72-78 (2000)

[文献書誌] I.Royal: "Activation of cdc42,rac, PAK, and rho-kinase in response to hepatocyte growth factor differentially regulates epithelial cell colony spreading and dissociation"Mol.Biol.Cell. 11. 1709-1725 (2000)

[文献書誌] N.Inagaki: "Activation of Ca^<2+>/calmodulin-dependent protein kinase II within post-synaptic dendritic spines of cultured hippocampal neurons"J.Biol.Chem.. 275. 27165-27171 (2000)

[文献書誌] M.Sone: "Synaptic development is controlled in the periactive zones of drosophila synapses"Development.. 127. 4157-4168 (2000)

[文献書誌] T.Kandabashi: "Inhibition of myosin phosphatase by upregulated rho-kinase plays a key role for coronary artery spasm in a porcine model with interleukin-1beta"Circulation. 101. 1319-1323 (2000)

[文献書誌] S.Miyamoto: "Changes in E-cadherin associated with cytoplasmic molecules in well and poorly differentiated endometrial cancer"Br.J.Cancer. 83. 1168-1175 (2000)

[文献書誌] K.Ihara: "Crystallization and preliminary crystallographic analysis of the rho-binding domain of bovine rho-kinase"Acta.Crystallogr. D.Biol.Crystallogr.. 56. 1042-1044 (2000)

[文献書誌] P.A.Oude Weernink: "Stimulation of phosphatidylinositol-4-phosphate 5-kinase by Rho-kinase"J.Biol.Chem.. 275. 10168-10174 (2000)

[文献書誌] K.Szaszi: "RhoA and rho kinase regulate the epithelial Na^+/H^+ exchanger NHE3. Role of myosin light chain phosphorylation"J.Biol.Chem.. 275. 28599-28606 (2000)

[文献書誌] R.Hashimoto: "Site-specific phosphorylation of neurofilament-L is mediated by calcium/calmodulin-dependent protein kinase II in the apical dendrites during long-term Potentiation"J.Neurochem.. 75. 373-382 (2000)

[文献書誌] M.Kanai-Azuma: "Co-localization of FAM and AF-6, the mammalian homologues of Drosophila faf and canoe, in mouse eye development"Mech.Dev.. 91. 383-386 (2000)

[文献書誌] T.Hamaguchi: "Phosphorylation of CPI-17, an inhibitor of myosin phosphatase, by protein kinase N"Biochem. Biophys. Res.Commun.. 274. 825-830 (2000)

[文献書誌] Y.Fukata: "Rho-Rho-kinase pathway in smooth muscle contraction and cytoskeletal reorganization of non-muscle cells"TRENDS in Pharm.Sci.. 22. 32-39 (2001)

[文献書誌] M.Fukata: "Involvement of IQGAP1, an Effector of Rac1 and Cdc42 GTPases, in Cell-Cell Dissociation during Cell Scattering"Mol.Cell.Biol.. 21(in press). (2001)

2000 年度実績報告書

貝淵弘三名古屋大学, 医学部, 教授 (00169377)