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Intracellular and intercellular signaling mediated by Eph tyrosine kinase receptor in vascular endothelial cells

Research Project

Project/Area Number 14380342
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionNational Cardiovascular Center Research Institute

Principal Investigator

MOCHIZUKI Naoki  National Cardiovascular Center Research Institute, Structural Analysis, Director, 循環器形態部, 部長 (30311426)

Co-Investigator(Kenkyū-buntansha) ONISHI Hirofumi  National Cardiovascular Center Research Institute, Structural Analysis, Section chief, 循環器形態部, 室長 (80092542)
MASUDA Michitaka  National Cardiovascular Center Research Institute, Structural Analysis, Section chief, 循環器形態部, 室長 (00190364)
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2004: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥7,800,000 (Direct Cost: ¥7,800,000)
Keywordssignal transduction / tyrosine kinase / receptor / cell-cell adhesion / angiogenesis / 情報伝達系
Research Abstract

In this research project, we have identified a Rho guanine nucleotide exchange factor(GEF)espetially expressed in vascular smooth muscle cells. We named this exchange factor Vsm-RhoGEF(Vascular smooth muscle specific GEF) after its expression and characterized the exchange specificity and localization. Vsm-RhoGEF functions downstream of EphA4 receptor and is expressed exclusively in the arterial smooth muscle cells (VSMCs), where EphA4 receptor is localized/. Furtheremore, Vsm-RhoGEF exhibits GEF activity for Rac as well as Rho. Consistently, when VSMCs were stimulated with ephrin-A1,they showed remarkable membrane ruffling, which is a hallmark of Rac activation. Vsm-RhoGEF is localized on actin stress fiber in unstimulated VSMCs, whereas it is dissociated from stress fiber to peripheral ruffled membrane upon ephrin-A1 simulation. Thus, while the Vsm-RhoGEF on stress fiber may function as a RhoGEF for Rho-RhoKinase signaling to regulate acttin-myosin coupling, the dissocicated Vsm-RhoG … More EF activates Rac for membrane extension.
We further investigated the Ras family GTPases, especially R-Ras family functioning downstream of Eph tyrosine kinase. R-Ras has been suggested to function for extracellular matrix-cell adhesion when simulated by ephrin. We hypothesized that actomyosin is regulated by R-Ras, because MysoinIX contains Ras binding domain in its ajnino-terminus of the myosin head. R-Ras family consists of R-Ras, M-Ras, and TC21. We found that among them R-Ras and TC21 preferentially bind to myosinIX. Other Ras family members, H-Ras, Rap1 and Ral do not bind to MysoinIX. Thus Eph-activated R-Ras and TC21 may promote myosinIX-based motor function such as vesicular frafficking or muscle contraction.
Eph tyrosine kinase receptor is endocytosed similarly to other tyrosine kinase receptor family members. We first explored the endocytosis of carboxy-terminally EGFP-tagged EphB receptors upon ephrin-B1 stimulation. PECAM-1-EGFP was used as a negative control. The cells expressing either EphB1-EGFP or PECAM-1-EGFP stimulated with ephrin-B1 were time-lapse imaged. We found that EphB1-EGFP exhibited the oligomerization upon stimulation while PECAM-1 EGFP did not. In addition to oligomerization, we found that oligomerized EphB1-EGFP was endocytosed into the cell body f from the plasma membrane. These results suggest that Eph-ephrin-mediated intracellular signaling is triggerd upon cell-cell contact and modulated by endocytosis. We will prceed to examine the molecular mechanism how Vsm-RhoGEF functions as GEF switch for Rho and/or Rac. Less

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (39 results)

All 2005 2004 2003 2002 Other

All Journal Article (31 results) Publications (8 results)

  • [Journal Article] Cyclic AMP potentiates VE-cadherin-mediated cell-cell contact to enhance endothelial barrier function through an Epac-Rap1 signaling pathway.2005

    • Author(s)
      Fukuhara S, et al.
    • Journal Title

      Mol.Cell.Biol. 25

      Pages: 133-146

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Cyclic AMP potenciates VE-cadherin-mediated cell-cell contact to enhance endothelial berrier function through an Epac-Rap1 signaling pathway.2005

    • Author(s)
      Fukuhara S., et al.
    • Journal Title

      Mol.Cell.Biol. 25

      Pages: 133-146

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Cyclic AMIP potenciates VE-cadherin-mediated cell-cell contact to enhance endothelial barrier function through an Epac-Rap1 signaling pathway.2005

    • Author(s)
      Fukuhara S, et al.
    • Journal Title

      Mol.Cell.Biol. 25

      Pages: 133-146

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Identification and characterization of zipper-interacting protein kinase as the unique vascular smooth muscle myosin phosphatase-associated kinase.2004

    • Author(s)
      Endo A, et al.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 42055-42061

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Transmission of force and displacement within the myosin molecule.2004

    • Author(s)
      Ohki T, et al.
    • Journal Title

      Biochemistry 43

      Pages: 13707-13714

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] A novel dynamin-associating molecule, forming-binding protein 17, induces tubular membrane invaginations and participates in endocytosis.2004

    • Author(s)
      Kamioka Y, et al.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 40091-40099

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Cell Type-specific Regulation of RhoA Activity during Cytokinesis.2004

    • Author(s)
      Yoshizaki H, et al.
    • Journal Title

      J.Biol.Chem 279

      Pages: 44756-44762

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Regulatory roles for APJ, a seven-transmembrane receptor related to angiotensin-type 1 receptor in blood pressure in vivo2004

    • Author(s)
      Ishida J, et al.
    • Journal Title

      J.Biol.Chem 279

      Pages: 26274-26279

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Identification and characterization of zipper-interacting protein kinase as the unique vascular smooth muscle mvosin phosphatase-associated kinase.2004

    • Author(s)
      Endo A., et al.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 42055-42061

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Transmission of force and displacement within the myosin molecule.2004

    • Author(s)
      Ohki T., et al.
    • Journal Title

      Biochemistry. 43

      Pages: 13707-13714

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A novel dynamin-associating molecule, forming-binding protein 17,induces tubular membrance invaginations and participates in endocytosisi.2004

    • Author(s)
      Kamioka Y., et al.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 40091-40099

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Cell Type-specific Regulation of RhoA Activity during Cytokinesis.2004

    • Author(s)
      Yoshizaki H., et al.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 44756-44762

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Regulatory roles for APJ, a seven-transmembrance receptor related to angiotensin-type 1 receptor in bolld pressure in vivo.2004

    • Author(s)
      Ishida J., et al.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 26274-26279

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Cell Type-specific Regulation of RhoA Activity during Cytokinesis.2004

    • Author(s)
      Yoshizaki H, et al.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 44756-44762

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Regulatory roles for APJ, a seven-transmembrane receptor related to angiotensin-type 1 receptor in blood pressure in vivo2004

    • Author(s)
      Ishida J, et al.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 26274-26279

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Identification of fer tyrosine kinase localized on molecule-1 phosphorylating kinase in vascular endothelial cells.2003

    • Author(s)
      Endo A, et al.
    • Journal Title

      Mol.Biol.Cell. 14

      Pages: 3553-3564

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] EphA4-mediated Rho activation via Vsm-RhoGEF expressed specifically in vascular smooth muscle cells.2003

    • Author(s)
      Ogita H, et al.
    • Journal Title

      Circ.Res. 93

      Pages: 23-31

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Activity of Rho-family GTPases during cell division as visualized with FRET-based probes.2003

    • Author(s)
      Nagashima K, et al.
    • Journal Title

      J.Cell Biol. 162

      Pages: 1-10

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A Selective inhibition of vascular endothelial growth factor receptor-2(VEGFR-2)identifies a central role for VEGFR-2 in human aortic endothelila cell responses to VEGF.2003

    • Author(s)
      Fukuhara S, et al.
    • Journal Title

      J.Receptor Signal Transduction 23

      Pages: 239-254

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Hybrid cell-gene therapy for pulmonary hypertension based on phagocytosing action of endothelila progenitor cells.2003

    • Author(s)
      Nagaya K, et al.
    • Journal Title

      Circulation 108

      Pages: 889-895

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Identification of fer tyrosine kinase localized on molecule-1 phoshorylating kinase in vascular endothelial cells.2003

    • Author(s)
      Endo A., et al.
    • Journal Title

      Mol.Biol.Cell. 14

      Pages: 3533-3564

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] EphA4-mediated Rho activation via Vsm-RhoGFE expressed specifically in vascular smooth muscle cells.2003

    • Author(s)
      Ogita H., et al.
    • Journal Title

      Circ.Res. 93

      Pages: 23-31

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Activity of Rho-family GTPases during cell division as visualized with FRET-based probes.2003

    • Author(s)
      Nagashima K., et al.
    • Journal Title

      J.Cell Biol. 162

      Pages: 1-10

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A Selective inhibition of vascular endothelial growth factor receptor-2(VEGFR-2)indentifies a central role for VEGFR-2 in human aortic endothelial cell responses to VEGF.2003

    • Author(s)
      Fukuhara S., et al.
    • Journal Title

      J.Receptor Signal Transduction 23

      Pages: 239-254

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Hybrid cell-gene therapy for pulmonary hypertension based on pyagocytosing action of endothelial progenitor cells.2003

    • Author(s)
      Nagaya K, et al.
    • Journal Title

      Circulation. 108

      Pages: 889-895

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Adaptor protein Crk is required for ephrin-B1-induced membrane ruffling and focal complex assembly of human aortic endothelial cells.2002

    • Author(s)
      Nagashima KI, et al.
    • Journal Title

      Mol.Biol.Cell. 13

      Pages: 4231-4242

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Sphingosine 1-phosphate induces membrane ruffling and increases motility of human umbilical vein endothelial cells via vascular endothelial growth factor receptor and CrkII.2002

    • Author(s)
      Endo A, et al.
    • Journal Title

      J Biol Chem. 277

      Pages: 23747-23754

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] SWAP-70 is a guanine-nucleotide-exchange factor that mediates signalling of membrane ruffling.2002

    • Author(s)
      Shinohara M, et al.
    • Journal Title

      Nature 416

      Pages: 759-763

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Adaptor protein Crk is required for ephrin-B1-induced membrane ruffling And focal complex assembly of human aortic endothelial cells.2002

    • Author(s)
      Nagashima KI, et al.
    • Journal Title

      Mol.Biol.Cell. 13

      Pages: 4231-4242

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Sphingosine 1-phosphate induces membrane ruffling and increases umotility of human umbilical vein endothelial cells via vascular endothelial Growth factor receptor and CrkII.2002

    • Author(s)
      Endo A., et al.
    • Journal Title

      J.Biol.Chem. 277

      Pages: 23747-23754

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] SWAP-70 is a guanine-nucleotide-exchange factor that mediates signaling of membrane ruffling.2002

    • Author(s)
      Shinohara M., et al.
    • Journal Title

      Nature 416

      Pages: 759-763

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Endo A, et al.: "Identification of fer tyrosine kinase localized on molecule-1 phosphorylating kinase in vascular endothelial cells."Mol.Biol.Cell.. 14. 3553-3564 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ogita H, et al.: "EphA4-mediated Rho activation via Vsm-RhoGEF expressed specifically in vascular smooth muscle cells."Circ.Res.. 93. 23-31 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nagashima K, et al.: "Activity of Rho-family GTPases during cell division as visualized with FRET-based probes."J.Cell Biol.. 162. 1-10 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Fukuhara S, et al.: "A Selective inhibition of vascular endothelial growth factor receptor-2(VEGFR-2)identifies a central role for VEGFR-2 in human aortic endothelila cell responses to VEGF."J.Receptor Signal Transduction.. 23. 239-254 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nagaya K, et al.: "Hybrid cell-gene therapy for pulmonary hypertension based on phagocytosing action of endothelila progenitor cells."Circulation. 108. 889-895 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nagashima KI, et al.: "Adaptor protein Crk is required for ephrin-B1-induced membrane ruffling and focal complex assembly-of human aortic endothelial cells"Mol. Biol. Cell.. 13. 4231-4242 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Endo A, et al.: "Sphingosine 1-phosphate induces membrane ruffling and increases motility of human umbilical vein endothelial cells via vascular endothelial growth factor receptor and CrkII"J Biol Chem.. 277. 23747-23754 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shinohara M, et al.: "SWAP-70 is a guanine nucleotide-exchange factor that mediates signalling of membrane ruffling"Nature. 416. 759-763 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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