1999 Fiscal Year Annual Research Report

蛋白質の核膜通貨の分子メカニズム:ATPの作用と分子間相互作用の解析

Research Project

Project/Area Number	11480209
Research Category	Grant-in-Aid for Scientific Research (B)
Research Institution	Osaka University
Principal Investigator	今本尚子大阪大学, 医学系研究科, 助手 (20202145)
Co-Investigator(Kenkyū-buntansha)	米田悦啓大阪大学, 医学系研究科, 教授 (80191667)
Keywords	核タンパク質輸送 / 核膜孔複合体 / βーカテニン / importinβ / (低分子量GTPase)Ran / ATP
Research Abstract	輸送担体importinβは、核膜孔を通過する分子の中で、最も機能解析と構造解析が進められている。分子が核膜孔を通過するための分子間相互作用を知るため、核-細胞質間をシャトルするimportinβ単独の結晶構造を明らかにし、核から細胞質に出るが、細胞質から核の中には入らないimportinβ/Ran-GTP複合体、及び、細胞質から核内に入るimportinβ/IBB複合体の結晶構造と比較した。その結果、importinβは核膜孔複合体と相互作用する特定の領域で、構造変化を起こすflexibleな分子であることが明らかになった。こうした結晶構造に基づいて点変異をいれることにより、核膜孔通過能が低下するimportinβの変異蛋白を得ることができれば、通過する際に相互作用する核膜孔複合体を同定するのに有用なプローブとなることが考えられる。また、核膜孔をシャトルするβ-カテニンの、細胞質から核内に移行するために必要な領域と、核から細胞質に移行するために必要な領域を同定したところ、どちらも、10-12番目のアルマジロrepeat構造であることが明らかになった。importinβ単体の結晶構造と、βーカテニンの結晶構造を比較したところ、importinβが核膜孔を通過するのに必要な領域と、β-カテニンが10-12番目のアルマジロ構造で直接核膜孔複合体と相互作用することが示唆された。

Research Products

(12 results)

All Other

All Publications (12 results)

[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)
[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)
[Publications] Yokoya,F.: "β-Catenin can be transported into the nucleus in a Ran-unassisted manner"Mol.Biol.Cell. 10. 1119-1131 (1999)
[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)
[Publications] Sahara,S.: "Acinus, a caspase-3-activated protein responsible for apoptotic chromatin condensation"Nature. 401. 168-173 (1999)
[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)
[Publications] Kose,S.: "Distinct energy requirement for nuclear import and export of importinβ in living cells"FEBS Lett.. 463. 327-330 (4999)
[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)
[Publications] Matsubae,M.: "Characterization of the nuclear transport of a novel leucine-rich acidic nuclear protein-like protein"FEBS Lett. (in press).
[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).
[Publications] 小瀬真吾: "実験医学"羊土社. 7 (1999)
[Publications] 横屋史彦: "秀潤社"細胞工学. 7 (1999)

1999 Fiscal Year Annual Research Report

蛋白質の核膜通貨の分子メカニズム:ATPの作用と分子間相互作用の解析

Principal Investigator

今本 尚子 大阪大学, 医学系研究科, 助手 (20202145)

Research Products

[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)

[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)

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

[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)

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

[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)

[Publications] Kose,S.: "Distinct energy requirement for nuclear import and export of importinβ in living cells"FEBS Lett.. 463. 327-330 (4999)

[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)

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

[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).

[Publications] 小瀬 真吾: "実験医学"羊土社. 7 (1999)

[Publications] 横屋 史彦: "秀潤社"細胞工学. 7 (1999)

今本尚子大阪大学, 医学系研究科, 助手 (20202145)

[Publications] 小瀬真吾: "実験医学"羊土社. 7 (1999)

[Publications] 横屋史彦: "秀潤社"細胞工学. 7 (1999)