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Mehanism of molecular translocation through nuclear pore complex

Research Project

Project/Area Number 11480209
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionNatiomal Institute of Genetics (2000-2001)
Osaka University (1999)

Principal Investigator

IMAMTO Naoko  Natiomal Institute of Genetics, Structural Biology Center, 構造遺伝学研究センター, 助教授 (20202145)

Co-Investigator(Kenkyū-buntansha) YONEDA Yoshihiro  Osaka University Graduate School of Medicine Professor, 大学院・医学系研究科, 教授 (80191667)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥15,300,000 (Direct Cost: ¥15,300,000)
Fiscal Year 2000: ¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1999: ¥8,600,000 (Direct Cost: ¥8,600,000)
KeywordsNuclear Transport / Nuclear Pore Complex / Importin β / β-catenin / Ran / ATP / Crystal Stracture / importin β / 核タンパク質輸送 / βーカテニン / (低分子量GTPase)Ran
Research Abstract

In contrast to our growing knowledge of receptor molecules for nucleocytoplasmic transport, function of nuclear pore complex (NPC), as well as mechanism of translocation step of transport is still very poorly understood. In this study, we focused on the following study. (1) Structural study of importin p. We solved crystal structure of uncomplexed form of importin β. Precise structurall comparison of our structur and previously reported structure of complexed form of importin β has revealed the flexible nature of importin β. Since local conformation change and flexible motion of importin β resides within its NPC- binding domain in which we have determined by series of deletion mutants, this flexible nature could be important for its movement through nuclear pores. We are now making series of point mutants of importin p that behave differently at NPC. (2) In order to study mechanism of a directional movement of importin β at NPC, we examined a requirement ofATP in import and export of importin p, and found that only nuclear export of importin β involves energy-requiring step(s) in living cells. In vitro transport assay revealed that an energy-dependent nuclear export of importin β requires soluble factors. To address the issue of how the energy is utilized during the NPC-passage of importin β, we are now attempting to purify an energy-dependent nuclear export factor of importin β from mouse Ehrlich cytosolic extract.

Report

(3 results)
  • 2001 Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (48 results)

All Other

All Publications (48 results)

  • [Publications] Chang-Jie Jiang: "Molecular Cloning of a Novel Importin α Homologue from Rice, by which constitutive photomorphogenic 1 (COP1z) nuclear localization signal (NLS)-orotein is preferentially nuclear imported"J. Biol. Chem.. 276. 9322-9329 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Soo Jae Lee: "The adoption of a twisted structure of importin-β is essential for the protein-protein interaction required for nuclear transport"J. Mol. Biol.. 302. 251-264 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Naoko Imamoto: "Diversity in nucleocytoplasmic transport pathways"Cell Structure and Function. 25. 207-216 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Taro Tachibana: "Recycling of importin α from the nucleus is suppressed by loss of RCC1 function in liying mammalian cells"Cell Structure and Function. 25. 115-123 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Masami Mathubae: "Characterization of the nuclear transport of a novel leucine-rich aoidic nuclear protein-like protein"FEBS Lett.. 468. 171-175 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tokuko Haraguchi: "Live fluorescence imaging reveals early recruitment of emerin, LBR, RanBP2, and Nup153 to reforming functianal nuclear envelopes"J. Cell Sci.. 113. 779-794 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shingo Kose: "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
      2001 Final Research Report Summary
  • [Publications] Takayuki Ohshima: "CRM1 mediates nuclear export of nonstructural protein 2 from parvovirus minute virus of mice"Biochem. Biophys. Res. Commun.. 264. 144-150 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shingo Kose: "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)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sethuko Sahara: "Acinus is a caspase-3-activated protein required for apoptotic cbromatin condensation"Nature. 401. 168-173 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Emi Nagoshi: "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 with HLH-Zip"Mol. Biol. Cell. 10. 2221-2233 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Fumihiko Yokoya: "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
      2001 Final Research Report Summary
  • [Publications] Miki Hieda: "A monoclonal antibody to the COOH-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
      2001 Final Research Report Summary
  • [Publications] 今本尚子: "蛋白質 核酸 酵素"核と細胞質の機能分子輸送 145巻. 2369-2377 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 今本尚子: "バイオサイエンスの新世紀"核-細胞質間分子輸送-細胞の司令塔をさらに管理するのは? 2巻. 122-136 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 小瀬真吾: "実験医学"細胞質から核へのタンパク質輸送の分子メカニズム 17巻. 2354-2360 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Chang-Jie Jiang, Kazuhiro Shoji, Rikyu Matsuki, Akihiki Baba, Noritoshi Inagaki, Hiroshi Ban, Toshisuke Iwasaki, Naoko Imamoto, Yoshihiro Yoneda, Xing-Wang Deng, and Naoki Yamamoto.: "Molecular Cloning of a Novel Importin a Homologue from Rice, by which constitutive photomorphogenic 1 (COP1) nuclear localization signal (NLS)-protein is preferentially nuclear imported.."J. Biol. Chem. 276. 9322-9329 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Soo Jae Lee, Naoko Imamoto, Hiroaki Sakai, Atsushi Nakagawa, Shingo Kose, Makiko Koike, Masaki Yamamoto, Takashi Kumasaka, Yoshihiro Yoneda and Tomitake Tsukihara: "The adoption of a twisted structure of importin-p is essential for the protein-protein interaction required for nuclear transport."J. Mol. Biol. 302. 251-264 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Naoko Imamoto: "Diversity in nucleocytoplasmic transport pathways"Cell Structure and Function. 25. 207-216 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Taro Tachibana, Miki Hieda, Yoichi Miyamoto, Shingo Kose, Naoko Imamoto, and Yoshihiro Yoneda: "Recycling of importin oc from the nucleus is suppressed by loss ofRCC1 function in living mammalian cells"Cell Structure and Functions. 25. 115-123 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Masami Matsubae, Toshinao Kurihara, Taro Tachibana, Naoko Imamoto and Yoshihiro Yoneda: "Characterization of the nuclear transport of a novel leucinerich acidic nuclear protein-like protein."FEES Lett. 468. 171-175 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tokuko Haraguchi, Takao Koujin, Tomohiro Hayakawa, Toru Kaneda, Chihiro Tsutumi, Naoko Imamoto, Chihiro Akazawa, Jun Sukegawa, Yoshihiro Yoneda, and Yasushi Hiraoka: "Live fluorescence imaging reveals early recruitment of emerin, LBR, RanBP2, and Nupl53 to reforming functional nuclear envelopes"J. Cell Sci. 113. 779-794 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shingo Kose, Naoko Imamoto, and Yoshihiro Yoneda: "Distinct energy requirement for nuclear import and export of importin P in living cells"FEES Lett. 463. 327-330 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Takayuki Ohshima, Toshihiro Nakajima, Takayuki Oishi, Naoko Imamoto, Yoshihiro Yoneda, Akiyoshi Fukamizu, Ken-ichi Yagami: "CRMl mediates nuclear export of nonstructural protein 2 from parvovirus minute virus of mice."Biochem. Biophys. Res. Commun.. 264. 144-50 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shingo Kose, Naoko Imamoto, Taro Tachibana, Minoru Yoshida, and Yoshihiro Yoneda: "β-Subunit of nuclear pore-targeting complex (Importin-p) can be exported from the nucleus in a Ran-independent manner."J. Biol. Chem.. 274. 3946-3952 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Setsuko Sahara, Mamoru Aoto, Yutaka Eguchi, Naoko Imamoto, Yoshihiro Yoneda, Yoshihide Tsujimoto: "Acinus is a caspase-3-activated protein required of apopototic chromatin condensation"Naturey. 401. 168-173 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Emi Nagoshi, Naoko Imamoto, Ryuichiro Sato and Yoshihiro Yoneda.: ".Nuclear impot of sterol regulatory element-binding protein-2, a basic helix-loop-helixItnpert eucine zipper (bHLH-Zip)-containing transcription factor, occurs through the direct interaction of importin P with HLH-Zip."Mol. Biol. Cell,. 10. 2221-2233 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Fumihiko Yokoya, Naoko Imamoto, Taro Tachtbana and Yoshihiro Yoneda: "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
      2001 Final Research Report Summary
  • [Publications] Miki Hieda, Taro Tachibana, Fumihiko Yokoya, Shingo Kose, Naoko Imamoto and Yoshihiro Yoneda.: "A monoclonal antibody to the COOH-terminal acidic Pcrtion of Ran inhibits both the recycling of Ran and nuclear protein import in iving cells'"J. Cell Biol. 144. 645-655 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Lee,S.J.: "The adoption of a twisted structure of importin βis essential for the protein-protein interacion required for nuclear transport."J.Mol.Biol.. 302. 251-264 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tachibana,T.: "Recycling of importinα from the nucleus is suppressed by loss of RCC1 function in living mammalian cells."Cell Struct.Funct.. 25. 115-123 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Matsubae,M.: "Characterization of the nuclear transport of a novel leucine-rich acidic protein-like protein."FEBS Left.. 468. 171-175 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Haraguchi,T.: "Reformation of the functional nuclear envelope during telophase : Tenporal sequences determined by fluorescence imaging in living Hela Cells."J.Cell Sci.. 113. 779-794 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Imamoto,N.: "Diversity in nucleocytoplasmic transport pathways"Cell Struct.Funct.. 25. 207-216 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 今本尚子: "蛋白質・核酸酵素"共立出版. 9 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 今本尚子: "バイオサイエンスの新世紀第2巻「タンパク質の一生」"共立出版. 15 (2000)

    • Related Report
      2000 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] 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] Nahoshi,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 βwith HTH-Zip"Mol.Biol.Cell. 10. 2221-2233 (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] Ohsima,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 importinβ in living cells"FEBS Lett.. 463. 327-330 (4999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Haragchi,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 function in living mammalian cells"Cell Struct Funct.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] 小瀬 真吾: "実験医学"羊土社. 7 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 横屋 史彦: "秀潤社"細胞工学. 7 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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