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Dynamics of nuclear shuttling with MAP kinase

Research Project

Project/Area Number 12480202
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionNagoya City University

Principal Investigator

NAKANISHI Mamoru  Nagoya City University, Graduate School of Pharmaceutical Sciences, Professor, 大学院・薬学研究科, 教授 (90090472)

Co-Investigator(Kenkyū-buntansha) FURUNO Tadahide  Nagoya City University, Graduate School of Pharmaceutical Sciences, Assistant Professor, 大学院・薬学研究科, 講師 (80254308)
HIRASHIMA Naohide  Nagoya City University, Graduate School of Pharmaceutical Sciences, Assistant Professor, 大学院・薬学研究科, 助教授 (10192296)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2001: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2000: ¥2,900,000 (Direct Cost: ¥2,900,000)
KeywordsMAP kinase / MAP kinase cascade / GFP / Confocal microscope / Fluorescent protein / Raf-1 / Nuclear shuttling / Basophile / キメラ蛋白質 / 核シヤトル
Research Abstract

The mitogen-activated protein (MAP) kinase cascade consists of MAP kinase (MAP; ERK2) and its activator, MAP kinase (MAPKK; MEK). However, the mechanisms for activation of ERK2 have not been defined yet in cells. Here, we used fluorescent-tagged ERK2 and MEK to examine the localization of ERK2 and MEK in living rat basophilic leukemia (RBL-2H3) cells. ERK2 was mainly in the cytoplasm in resting cells, but translocated into the nucleus after the ligation of IgE receptors. The import of ERK2 reached the maximum at 6-7 min, and then the imported ERK2 was exported from the nucleus. MEK mainly resided in the cytoplasm, and no significant MEK translocation was detected after ligation of IgE receptors. The data suggested that the sustained calcium increase was required for the optimal translocation of ERK2 into the nucleus in RBL cells. However, analysis of the dynamics of ERK2 and MEK suggested that both of them rapidly shuttle between the cytoplasm and the nucleus, and that MEK regulate the … More nuclear shuttling of ERK2 while MEK remain mainly in the cytoplasm. Pretreatment of RBL-2H3 cells with cytochalasin D and Latrunculin A, inhibitors of actin polymerization, prolonged the nuclear translocation of ERK2 after antigen stimulation. Western blotting analysis revealed that these drugs enhanced the phosphorylation of both ERK2 and MEK after antigen stimuation. To the contrary, nocodazole, an inhibitor of tubulin polymerization, caused neither prolongation of nuclear translocation of ERK2 nor enhancement of phosphorylation of ERK2 and MEK. Dissociation of cross-linked receptors by the addition of excess amount of monovalent hapten halted translocation of ERK2 even in cells pretreated with cytochalasin D. Taken together, these results indicate that actin polymerization affects the nuclear shuttling of ERK2 by the regulation of IgE receptor cross-linking. These results gave a new insight of the dynamics of ERK2 and MEK in the nuclear shuttling of RBL-2H3 cells after Ag stimulation. Less

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (33 results)

All Other

All Publications (33 results)

  • [Publications] Hasegawa, S., Hirashima, N., Nakanishi, M.: "Comparative Study of Transfection Efficiency of Cationic Cholesterols Mediated by Liposomes-Based Gene Delivery"Bioorg. Med. Chem. Lett.. 12. 1299-1302 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kato, N., Nakanishi, M., Hirashima, N.: "Transbilayer Asymmetry of Phospholipids in the Plasma Membrane Regulates Exocytotic Release in Mast Cells"Biochem.. 41. 8068-8074 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Noguchi, A., Hirashima, N., Furuno, T., Nakanishi, M.: "Activation of Receptor Tyrosine Kinases Promotes Gene Transfection in Rat Neuronal PC12 Cells by Cationic Liposomes"Neurosci. Lett.. 325. 29-32 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Noguchi, A., Hirashima, N,.Nakanishi, M.: "Cationic Choresterol Promotes Gene Transfection Using the Nuclear Localization Signal in Protamine"Phamaceut. Res.. 19. 933-938 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Mori, N., Suzuki, R., Furuno, T., McKay, DM.Wada, M., Teshima, R., Bienenstock, J., Nakanishi, M.: "Nerve-mast cell (RBL) interaction : RBL membrane ruffling occurs at the contact site with an activated neurite"Am J Physiol Cell Physiol.. 283. C1738-C1744 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Noguchi, S., Hirashima, N., Furuno, T., Nakanishi, M.: "Remarkable induction of apoptsis in cancer cells by a novel cationic liposome complexed with bcl-2 antisense oligonucleotide"J Control Release. 7. 313-320 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 中西 守 他: "免疫学辞典"東京化学同人. 450 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hasegawa,S., Hirashima,N., Nakanishi,M.: "Comparative Study of Transfection Efficiency of Cationic Cholesterols Mediated by Liposomes-Based Gene Delevery"Bioorg. Med. Chem. Lett.. 12. 1299-1302 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kato,N., Nakanishi,M., Hirashima,N.: "Transbilayer Asymmetry of Phospholipids in the Plasma Membrane Regugates Exocytotic Release in Mast Cells"Biochem.. 41. 8068-8074 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Noguchi,A., Hirashima,N., Furuno,T., Nakanishi,M.: "Activation of Receptor Tyrosine Kinases Promotes Gene Transfection in Rat Neuronal PC12 Cells by Cationic Liposomes"Neurosci. Lett.. 325. 29-32 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Noguchi,A., Hirashima,N., Nakanishi,M.: "Cationic Cholesterol Promotes Gene Transfection Using the Nuclear Localization Signal in Protamine"Pharmaceut. Res.. 19. 933-938 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Mori,N., Suzuki,R., Furumoto,T., McKay,DM., Wada,M., Teshima,R., Bienenstock,J., Nakanishi,M.: "Nerve-mast cell (RBL) interaction: RBL membrane ruffling occurs at the contact site with an activated neurite"Am J Physiol Cell Physiol. 283. C1738-1744 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Noguchi,S., Hirashima,N., Furuno,T., Nakanishi,M.: "Remarkable induction of apoptosis in cancer cells by a novel cationic liposome conplexed with a bcl-2 antisense oligonucleotide"J Control Release. 7. 313-320 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hasegawa, S., Hirashima, N., Nakanishi, M.: "Comparative Study of Transfection Efficiency of Cationic Cholesterols Mediated by Liposomes-Based Gene Delivery"Bioorg. Med. Chem. Lett.. 12. 1299-1302 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kato, N., Nakanishi, M., Hirashima, N.: "Transbilayer Asymmetry of Phospholipids in the Plasma Membrane Regulates Exocytotic Release in Mast Cells"Biochem.. 41. 8068-8074 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Noguchi, A., Hirashima, N., Furuno, T., Nakanishi, M.: "Activation of Receptor Tyrosine Kinases Promotes Gene Transfection in Rat Neuronal PC12 Cells by Caitonic Liposomes"Neurosci. Lett.. 325. 29-32 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Noguchi, A., Hirashima, N., Nakanishi, M.: "Cationic Choresterol Promotes Gene Transfection Using the Nuclear Localization Signal in Protamine"Pharmaceut. Res.. 19. 933-938 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Mori, N., Snzuki, R., Furuno, T., McKay, DM., Wada, M., Teshima, R., Bienenstock, J., Nakanishi, M.: "Nerve-mast cell (RBL) interaction : RBL membrane ruffling occurs at the contact site with as activated neurite"Am J Physiol Cell Physiol.. 283. C1738-C1744 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Noguchi, S., Hirashiama, N., Furuno, T., Nakanishi, M.: "Remarkable induction of apoptsis in cancer cells by a novel cationic liposome complexed with a bcl-2 autisense oligonucleotide"J Control Release. 7. 313-320 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 中西 守他: "免疫学辞典"東京化学同人. 450 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Furuno, T., Hirashima, N., Onizawa, S., Sagiya, N., Nakanishi, M.: "Nuclear shuttling of mitogen-activated protein (MAP) kinase (extracellular signal-regulated kinase (ERK) 2) was dynamically controlled by MAP/ERK kinase after antigen stimulation in RBL-2H3 cells"J. Immunol.. 166. 4416-4421 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nakanishi, M., Noguchi, A.: "Confocal and probe microscopy to study gene transfection-mediated by cationic liposomes with a cationic cholesterol derivative"Adv. Drug Delivery Rev.. 52. 197-207 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hasegawa, S., Hirashima, N., Nakanisbi, M.: "Microtubule involvements in the intracellular dynamics for gene transfection mediated by cationic liposomes"Gene Therapy. 8. 1669-1673 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Inoh, Y, Kitamoto, D., Hirashima, N., Nakanishi, M.: "Biosurfactants of MEL-A increase gene transfection mediated by cationic liposomes"Biochem. Biophys. Res. Commun.. 289. 57-61 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Noguchi, A., Hirashima, N., Furuno, T., Nakankshi, M.: "Activation of receptor tyrosine kinases promotes gene transfection in the neuronal PC12 cells by cationic liposomes"Neurosci. Lett.. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] Noguchi, A., Hirashima, N., Nakanishi, M.: "A cationic cholesterol promote recognition of protamine having NLS in gene transfection"Pharmaceut. Res.. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] 中西守 他: "免疫学辞典"東京化学同人. 450 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Dace, A.,Zhao,L.,Park,K.S.,Furuno,T.,Takamura,N.,Nakanishi,M.,West,B., et al.: "Hormone binding induces rapid proteasome-mediated degradation of thyroid hormone receptors."Proc.Natl.Acad.Sci.USA.. 97. 8985-8990 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ohshiro,H.,Suzuki,R.,Furuno,T.,Nakanishi,M.: "Atomic force microscopy to study direct neurite-mast cell (RBL) communication in vitro."Immunol.Lett.. 74. 211-214 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Suzuki,R.,Furuno,T.,Teshima,R.,Nakanishi,M.: "Bi-directional flow of in vitro mast cell-nerve communication observed by confocal laser scanning microscopy."Biol.Pharm.Bull.. 24. 291-294 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Furuno,T.,Hirashima,N.,Onizawa,S.,Sagiya,N.,Nakanishi,M.: "Nuclear shutting of mitogen-activated protein (MAP) kinase (extracellular signal-regulated kinase (ERK) 2) was dynamically controlled by MAP/ERK kinase after antigen stimulation in RBL-2H3 cells."J.Immunol.. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Amano,T.,Furuno,T.,Hirashima,N.,Ohyama,N.,Nakanishi,M.: "Dynamics of intracellular granules with CD63-GFP in rat basophilic leukemia cells."J.Biochem.. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Nakamura,R.,Nakanishi,M.: "Atomic force microscopy to study the effects of ITIM-bearing Fc γ RIIB on the activation of RBL-2H3 cells"Immunol.Lett.. 72. 167-170 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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