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Analysis of neuron-specific nuclear protein transport by using CaM kinase IV as a substrate.

Research Project

Project/Area Number 10480200
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionOsaka University

Principal Investigator

YONEDA Yoshihiro  Graduate School of Medicine Osaka University, Professor, 医学系研究科, 教授 (80191667)

Co-Investigator(Kenkyū-buntansha) IMAMOTO Naoko  Graduate School of Medicine Osaka University, Assistant Professor, 医学系研究科, 助手 (20202145)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥12,800,000 (Direct Cost: ¥12,800,000)
Fiscal Year 1999: ¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 1998: ¥7,000,000 (Direct Cost: ¥7,000,000)
Keywordsnuclear protein import / nuclear localization signal / neuron / CaM kinase / semi-intact cell / importin / cell nucleus
Research Abstract

In order to know the molecular mechanism of neuron-specific nuclear protein transport, we used Ca^<2+>/calmodulin-dependent protein kinase type IV (CaM kinase IV) as a model substrate. CaM kinase IV is known to be localized in the nucleus of neuronal cells, while throughout the cytoplasm of non-neuronal cells such as parathyroid cells. Further, the nuclear localization signal of CaM kinase IV has not yet been identified. When the recombinant CaM kinase IV proteins were injected into the cytoplasm of HeLa cells (human cervical cancer cells) or COS7 cells (African green monkey kidney cells), they migrated into the nuclei of COS7 cells but not those of HeLa cells, suggesting that CaM kinase IV is translocated from cytoplasm to the nucleus in a cell-type specific manner. Next, we tried to identify factors required for the nuclear import of CaM kinase IV by using a permeabilized cell-free system. It was demonstrated that brain extracts support the nuclear import of CaM kinase IV more efficiently than Ehrlich ascites tumor cell extracts. Moreover, the import was not inhibited by the addition of IBB (importin β-binding) domain of importin α and the N-terminal NPC (nuclear pore complex)-binding portion of importin β, meaning that the nuclear migration of CaM kinase IV is not mediated by conventional importin α/β pathway. More interestingly, it was found that the nuclear import mediated by brain extracts was not inhibited by the treatment with wheat germ agglutinin, whereas was that by Ehrlich ascites tumor cell extracts, suggesting that CaM kinase IV may be transported into the nucleus through at least two independent pathways. In the near future, factors involved in these reactions should be isolated and characterized.

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (62 results)

All Other

All Publications (62 results)

  • [Publications] Tachibana,T.: "Up-regulation of nuclear protein import by nuclear localization signal sequences in living cells."FEBS Lett.. 442. 235-240 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuoka,Y.: "Identification and characterrization of nuclear pore subcomplexes in mitotic extract of human somatic cells."Biochem.Biophys.Res.Commun.. 254. 417-423 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kose,S.: "β-Subunit of nuclear pore-targeting complex (importin-β) can be exported from the nucleus in a Ran-indenpendent manner."J.Biol.Chem.. 274. 3946-3952 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Hieda,M.: "A monoclonal antibody to the C-terminal acidic portion of Ran inhibits both the recycling of Ran and nuclear protein import in living cells."J.Cell Biol.. 144. 645-655 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kamei,Y.: "Three distinct classes of the α subunit of the nuclear pore-targeting complex (importin-α) are differentially expressed in adult mouse tissues."J.Histochem.Cytochem.. 47. 363-372 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kikuchi,N.: "Molecular shape and ATP binding activity of rat p50, a putative mammalian homologue of RuvB DNA helicase."J.Biochem.. 125. 487-494 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yokoya,F.: "β-Catenin can be transported into the nucleus in a Ran-unassisted manner."Mol.Biol.Cell. 10. 1119-1131 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nagoshi,E.: "Nuclear import of sterol regulatory element-binding protein-2, a basic helix-loop-helix-leucine zipper (bHLH-Zip)-containing transcription factor occurs through the direct interaction of importin β with HTH-Zip"Mol.Biol.Cell. 10. 2221-2233 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takagi,M.: "Chmadrin : a novel Ki-67 antigen-related perichromosomal protein possibly implicated in higher order chromatin structure."J.Cell Sci.. 112. 2463-2472 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sahara,S.: "Acinus, a caspase-3-activated protein responsible for apoptotic chromatin condensation."Nature. 401. 168-173 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Ohshima,T.: "CRM1 mediates nuclear export of nonstructural protein 2 from parvovirus minute virus of mine."Biochem.Biophys.Res.Commun.. 264. 144-150 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kose,S.: "Distinct energy requirement for nuclear import and export of importin β in living cells."FEBS Lett.. 463. 327-330 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yoneda,Y.: "Nucleocytoplasmic protein transport and recycling of Ran."Cell Struct.Funct.. 24. 425-433 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Haraguchi,T.: "Live fluorescence imaging reveals early recruitment of emerin, LBR, HP1β and RanBP2 to reforming functional nuclear envelope."J.Cell Sci.. 113. 779-794 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsubae,M.: "Characterization of the nuclear transport of a novel leucine-rich acidic nuclear protein-like protein."FEBS Lett.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Techibana,T.: "Recycling of importin α from the nucleus is suppressed by loss of RCC1 fuction in living mammalian cells."Cell Struct Funct.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tachibana, T.: "Up-regulation of nuclear protein import by nuclear localization signal sequences in living cells."FEBS Lett.. 442. 235-240 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuoka, Y.: "Identification and characterrization of nuclear pore subcomplexes in mitotic extract of human somatic cells."Biochem.Biophys.Res.Commun.. 254. 417-423 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kose, S.: "β-Subunit of nuclear pore-targeting complex (importin-β) can be exported from the nucleus in a Ran-indenpendent manner."J.Biol.Chem.. 274. 3946-3952 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Hieda, M.: "A monoclonal antibody to the C-terminal acidic portion of Ran inhibits both the recycling of Ran and nuclear protein import in living cells."J.Cell Biol.. 144. 645-655 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kamei, Y.: "Three distinct classes of the α subunit of the nuclear pore-targeting complex (importin-α) are differentially expressed in adult mouse tissues."J.Histochem, Cytochem.. 47. 363-372 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kikuchi, N.: "Molecular shape and ATP binding activity of rat p50, a putative mammalian homologue of RuvB DNA helicase."J.Biochem.. 125. 487-494 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yokoya, F.: "β-Catenin can be transported into the nucleus in a Ran-unassisted manner."Mol.Biol.Cell. 10. 1119-1131 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nagoshi, E.: "Nuclear import of sterol regulatory element-binding protein-2, a basic helix-loop-helix-leucine zipper (bHLH-Zip )-containing transcription factor occurs through the direct interaction of importin β with HTH-Zip."Mol.Biol.Cell. 10. 2221-2233 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takagi, M.: "Chmadrin ; a novel ki-67 antigen-related perichromosomal protein possibly implicated in higher order chromatin structure."J.Cell Sci.. 112. 2463-2472 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sahara, S.: "Acinus, a caspase-3-activated protein responsible for apoptotic chromatin condensation."Nature. 401. 168-173 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Ohshirma, T.: "CRM1 mediates nuclear export of nonstructural protein 2 from parvovirus minute virus of mine."Biochem.Biophys.Res.Commun.. 264. 144-150 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kose, S.: "Distinct energy requirement for nuclear import and export of importin β in living cells."FEBS Lett.. 463. 327-330 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yoneda, Y.: "Nucleocytoplasmic protein transport and recycling of Ran."Cell Struct.Funct.. 24. 425-433 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Haraguchi, T.: "Live fluorescence imaging reveals early recruitment of emerin, LBR, HP1 β and RanBP2 to reforming functional nuclear envelope."J.Cell Sci.. 113. 779-794 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsubae, M.: "Characterization of the nuclear transport of a novel leucine-rich acidic nuclear protein-like protein."FEBS Lett.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tachibana, T.: "Recycling of importin α from the nucleus is suppressed by loss of RCC1 function in living mammalian cells."Cell Struct Funct.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tachibana, T.: "Up-regulation of nuclear protein import by nuclear localization signal sequences in living cells"FEBS Lett.. 442. 235-240 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Matsuoka, Y.: "Identification and characterization of nuclear pore subcomplexes in mitotic extract of human somatic cells"Biochem. Biophys. Res. Commun.. 254. 417-423 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kose, S.: "β-Subunit of nuclear pore-targeting complex(importin-β) can be exported from the nucleus in a Ran-independent manner"J. Biol. Chem.. 274. 3946-3952 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Hieda, M.: "A monoclonal antibody to the C-terminal acidic portion of Ran inhibits both the recycling of Ran and nuclear protein import in living cells"J. Cell Biol.. 144. 645-655 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kamei, Y.: "Three distinct classes of the α subunit of the nuclear pore-targeting complex(importune-α) are differentially expressed in adult mouse tissues"J. Histochem. Cytochem.. 47. 363-372 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kikuchi, N.: "Molecular shape and ATP binding activity of rat p50, a putative mammalian homologue of RuvB DNA helicase"J. Biochem.. 125. 487-494 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yokoya, F.: "β-Catenin can be transported into the nucleus in a Ran-unassisted manner"Mol. Biol. Cell. 10. 1119-1131 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nagoshi, E.: "Nuclear import of sterol regulatory element-binding protein-2, a basic helix-loop-helix-leucine zipper(bHLH-Zip)-containing transcription factor occurs throough the direct interaction of importin β with HTH-Zip"Mol. Biol. Cell. 10. 2221-2233 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takagi, M.: "Chmadrin: a novel Ki-67 antigen-related perichromosomal protein possibly implicated in higher order chromatin structure"J. Cell Sci.. 112. 2463-2472 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sahara, S.: "Acinus, a caspase-3-activated protein responsible for apoptotic chromatin condensation"Nature. 401. 168-173 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Ohshima, T.: "CRM1 mediates nuclear export of nonstructural protein 2 from parvovirus minute virus of mine"Biochem. Biophys. Res. Commun.. 264. 144-150 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kose, S.: "Distinct energy requirement for nuclear import and export of importin β in living cells"FEBS Lett.. 463. 327-330 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yoneda, Y.: "Nucleocytoplasmic protein transport and recycling of Ran"Cell Struct. Funct.. 24. 425-433 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Haraguchi, T.: "Live fluorescence imaging reveals early recruitment of emerin, LBR, HP1β and RanBP2 to reforming functional nuclear envelope"J. Cell Sci.. 113. 779-794 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Matsubae, M.: "Characterization of the nuclear transport of a novel leucine-rich acidic nuclear protein-like protein"FEBS Lett.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Tachibana, T.: "Recycling of importin α from the nucleus is suppressed by loss of RCC1 fuction in living mammalian cells"Cell Struct Funct.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Togo Ikuta: "Nuclear Localization and Export Signals of the Human Aryl Hydrocarbon Receptor" THE JOURNAL OF BIOLOGICAL CHEMISTRY. 273・5. 2895-2904 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] N.Imamoto: "Nuclear Transport Factors:Function,Behavior and Interaction" Eur.J.Histochem.42. 9-20 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Norihiko Nakamura: "Localization of the Apoptosis-inducing Activity of Lupus Anticoagulant in an Annexin V-binding Antibody Subset" J.Clin.Invest.101・9. 1951-1959 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Murai,MD: "Temporal lobe epilepsy in a genius of natural history MRI volumetric study of postmortem brain" NEUROLOGY. 50. 1373-1376 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Nobuaki Kudo: "Leptomycin B Inhibition of Signal-Mediated Nuclear Export by Direct Binding to CRM1" EXPERIMENTAL CELL RESEARCH. 242. 540-547 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takahiro Matsusaka: "Mutations in fission yeast Cut15,an importin α homolog,lead to mitotic progression without chromosome condensation" Current Biology. 8・18. 1031-1034 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Chang-Jie Jiang: "Functional Characterization of a Plant Importin α Homologue" THE JOURNAL OF BIOLOGICAL CHEMISTRY. 273・37. 24083-24087 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Chang-Jie Jiang: "In vitro characterization of rice importin β1:molecular interaction with nuclear transport factors and mediation of nuclear protein import" FEBS Letters. 437. 127-130 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masami Yamada: "Nuclear transport factor p10/NTF2 function as a Ran-GDP dissociation inhibitor(Ran-GDI)" Current Biology. 8. 1342 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yosuke Matsuoka: "Identificatin and Characterization of Nuclear Pore Subcomplexes in Mitotic Extract of Human Somatic Cells" Biochemical and Biophysical Research Communications. 254. 417-423 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Taro Tachibana: "Up-regulation of nuclear protein import by nuclear localization signal sequences in living cells" FEBS Letters. 442. 235-240 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shingo Kose: "β-Subunit of Nuclear Pore-targeting Complex(Importin-β)Can Be Exported from the Nucleus in a Ran-independent Manner" THE JOURNAL OF BIOLOGICAL CHEMISTRY. 274・7. 3946-3952 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yasuhiro Kamei: "Three Distinct Classes of the α-Subunit of the Nuclear Pore-targeting Complex(Importin-α)Are Differentially Expressed in Adult Mouse Tissues" The Journal of Histochemistry & Cytochemistry. 47・3. 1-10 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Fumihiko Yokoya: "β-Catenin Can Be Transported into the Nucleus in a Ran-unassisted Manner" Molecular biology of the Cell. (in press). (1999)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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