2000 Fiscal Year Annual Research Report

Rhoファミリーによる細胞骨格と細胞接着の制御機構

Research Project

Project/Area Number	11480211
Research Institution	Nagoya University
Principal Investigator	貝淵弘三名古屋大学, 医学部, 教授 (00169377)
Co-Investigator(Kenkyū-buntansha)	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)
Keywords	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
Research Abstract	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。

Research Products
(20 results)

All Other

All Publications (20 results)

[Publications] 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)
[Publications] N.Nakamura: "Phosphorylation of ERM proteins at filopodia induced by Cdc42"Genes Cells. 5. 571-581 (2000)
[Publications] M.Amano: "Regulation and functions of rho-associated kinase"Exp.Cell Res.. 261. 44-51 (2000)
[Publications] M.Amano: "Purification and in vitro activity of Rho-associated kinase"Methods Enzymol.. 325. 149-155 (2000)
[Publications] T.Kaneko: "Identification of calponin as a novel substrate of Rho-kinase"Biochem. Biophys. Res.Commun.. 273. 110-116 (2000)
[Publications] A.Kawajiri: "Identification of a novel beta-catenin-interacting protein"Biochem. Biophys. Res.Commun.. 273. 712-717 (2000)
[Publications] T.Izawa: "Elongation factor-lalpha is a novel substrate of rho-associated kinase"Biochem. Biophys. Res.Commun.. 278. 72-78 (2000)
[Publications] 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)
[Publications] 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)
[Publications] M.Sone: "Synaptic development is controlled in the periactive zones of drosophila synapses"Development.. 127. 4157-4168 (2000)
[Publications] 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)
[Publications] S.Miyamoto: "Changes in E-cadherin associated with cytoplasmic molecules in well and poorly differentiated endometrial cancer"Br.J.Cancer. 83. 1168-1175 (2000)
[Publications] 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)
[Publications] P.A.Oude Weernink: "Stimulation of phosphatidylinositol-4-phosphate 5-kinase by Rho-kinase"J.Biol.Chem.. 275. 10168-10174 (2000)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] T.Hamaguchi: "Phosphorylation of CPI-17, an inhibitor of myosin phosphatase, by protein kinase N"Biochem. Biophys. Res.Commun.. 274. 825-830 (2000)
[Publications] 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)
[Publications] 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 Fiscal Year Annual Research Report

Rhoファミリーによる細胞骨格と細胞接着の制御機構

Principal Investigator

貝淵 弘三 名古屋大学, 医学部, 教授 (00169377)

Research Products

[Publications] 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)

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

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

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

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

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

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

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

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

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

貝淵弘三名古屋大学, 医学部, 教授 (00169377)