核膜孔複合体の分子動態と核-細胞質間タンパク質輸送制御

2008 Fiscal Year Annual Research Report

• Project/Area Number: 16084204
• Research Institution: Osaka University
• Principal Investigator: 米田 悦啓 Osaka University, 生命機能研究科, 教授 (80191667)
• Keywords: 核膜孔複合体 / ヌクレオポリン / 核蛋白質輸送 / 原子間力顕微鏡 / 筋細胞分化 / 細胞核

Research Abstract

本研究課題では、細胞核の重要な高次構造の1つである核膜孔複合体に焦点を当て、この複合体の重要な機能である核-細胞質間蛋白質輸送の制御を指標にして、その分子構築ならびに分子動態を明らかにすることを目的として研究を行っている。本年度は、昨年度に引き続き、核膜孔のダイナミックな動態を解析するための実験系として、細胞分化の観点からの研究を進めた。具体的には、筋芽細胞であるマウスC2C12細胞を用い、細胞を未分化状態で増殖を続けさせた場合と分化誘導させた場合とで比較した場合、核膜孔複合体がどのように変化するか、また、変化するのであれば、その動態を個々のヌクレオポリンに着目して解析するとともに、発現量、局在が変化するヌクレオポリンがあるかどうかを解析した結果、NUP358の発現量が分化に応じて顕著に増加することがわかった。また、原子間力顕微鏡を用いて、核膜孔複合体の構造変化も解析したところ、分化に応じて、NUP358が局在していると思われる、核膜孔の細胞質側の直径が大きくなることを発見した。一方、富ロイシン型核外輸送シグナルを持つ蛋白質の核外輸送効率が、分化細胞では増加していることがわかった。さらに、RNAiの方法を用いて、NUP358の発現を低下させると、分化が抑制されることもわかった。これらの結果から、筋細胞分化に応じて、NUP358の発現が増加するとともに、核膜孔複合体の全体的な構造もダイナミックに変化し、機能的にも核内外の蛋白質輸送効率のバランスが変化して、分化に重要な役割をする因子の細胞質へ局在することが引き金となって分化が進行する可能性が示唆された。

Research Products

• [Journal Article] Infrared lager-Mediated gene induction in targeted single cells in vivo (2009)
  • Author(s): Kamei, Y., et.al.
  • Journal Title: Nature Methods 6 Pages: 79-81
  • Peer Reviewed
• [Journal Article] mDia2 shuttles between the nucleus and the cytoplasm throuigh the Importin-α/β- and CRM1-mediated nuclear transport meehanism (2009)
  • Author(s): Miki, T., et.al.
  • Journal Title: J. Biol. Chem 284 Pages: 99-104
  • Peer Reviewed
• [Journal Article] Adaptor Aly and co-adaptor Thoc5 function in the Tap-p15-mediated nuclear export of HSP70 mRNA (2009)
  • Author(s): Katahira, J., et.al.
  • Journal Title: EMBO J (in press)
  • Peer Reviewed
• [Journal Article] Selective localization of PCBP2 to cytoplasmic processing bodies (2009)
  • Author(s): Fujimura, K., et.al.
  • Journal Title: Biochim. Biophys. Acta-Molecular Cell Research (in press)
  • Peer Reviewed
• [Journal Article] Synergistic nuclear import of NeuroD1 and its partner transcription factor E47 via heterodimerization (2009)
  • Author(s): Mehmood, R., et.al.
  • Journal Title: Exp. Cell Res. (in press)
  • Peer Reviewed
• [Journal Article] Roles of the TREX complex in.nuclear export of mRNA (2009)
  • Author(s): Katahira, J. and Yoneda, Y
  • Journal Title: RNA Biol. (in press)
  • Peer Reviewed
• [Journal Article] The role of nuclear transport system in cell differentiation (2009)
  • Author(s): Yasuhara, N., et.al.
  • Journal Title: Seminars in Cell and Developmental Biology (in press)
  • Peer Reviewed
• [Journal Article] Generation and characterization of a monoclonal antibody against NPI-1 subfamily of importin α (2008)
  • Author(s): Tachibana, T., et.al.
  • Journal Title: Hybridoma 27 Pages: 285-289
  • Peer Reviewed
• [Journal Article] Localized regulation of axonal RanGTPase controls retrograde injury signaling in peripheral nerve (2008)
  • Author(s): Yudin, D., et.al.
  • Journal Title: Neuror 59 Pages: 241-252
  • Peer Reviewed
• [Journal Article] Individual binding pockets of importin β for FG-nucleoporins are diffierently regulated by RanGTP (2008)
  • Author(s): Otsuka, S., et.al.
  • Journal Title: Proc. Natl. Acad. Sci. USA 105 Pages: 16101-16106
  • Peer Reviewed
• [Presentation] C2C12細胞の筋分化における核膜孔複合体リモデリング (2009)
  • Author(s): Asally,M., Yasuda,Y., Oka, M., Otsuka, S., Yoshimura, S. H., Yorted_a, Y.
  • Organizer: 第26回染色体ワークショップ
  • Place of Presentation: 姫路
  • Year and Date: 20090126-28
• [Presentation] 核蛋白質輸送と生命機能 (2008)
  • Author(s): 米田悦啓
  • Organizer: BMB2008第31回日本分子生物学会年会・第81回日本生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 20081209-12
• [Presentation] 精子細胞形成過程におけるimportin alphaの核内機能 (2008)
  • Author(s): 宮本洋一、安田善也、山城朋子、永井理博、米田悦啓、David Jans、Kate Loveland
  • Organizer: BMB2008第31回日本分r子生物学会年会・第81回日本生化学会太会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 20081209-12
• [Presentation] Nuclear pore dynamics during skeletal myogenesis (2008)
  • Author(s): Asally, M., Yasuda, Y., Oka, M., Otsuka, S., Yoshimura, S. H., Yoneda, Y.
  • Organizer: Cold Spring Harbor Laboratory Meeting on Dynamic Organization of Nuclear Function
  • Place of Presentation: N. Y. /USA
  • Year and Date: 20080917-21
• [Presentation] Nuclear import mechanism of neuroD1, E47 and their heterodimeric complex (2008)
  • Author(s): Mehmood, R., Yasuhara, N., Oe, S., Nagai, M., Yoneda, Y
  • Organizer: Cold Spring Harbor Laboratory Meeting on Dynamic Organization of Nuclear Function
  • Place of Presentation: N. Y. /USA
  • Year and Date: 20080917-21
• [Presentation] The role of Nup98 in a Crml-mediated nuclear export (2008)
  • Author(s): Oka, M., Asally, M., Yasuda, Y., Yoneda, Y.
  • Organizer: Cold Spring Harbor Laboratory Meeting on Dynamic Organization of Nuclear Function
  • Place of Presentation: N. Y. /USA
  • Year and Date: 20080917-21
• [Presentation] Functional analysis of nuclear importin α accumulating in response to the stress (2008)
  • Author(s): Yasuda, Y., Miyainoto, Y., Yoneda, Y.
  • Organizer: Cold Spring Harbor Laboratory Meeting on Dynamic Organization of Nuclear Function
  • Place of Presentation: N. Y. /USA
  • Year and Date: 20080917-21
• [Presentation] Nucleo-cytoplasmic transport and the assembly of stress granules (2008)
  • Author(s): Ken Fujimura, Jun Katahira, Fumi Kano, Yoshihiro Yoneda, Masayuki Murata
  • Organizer: Cold Spring Harbor Laboratory Meeting on Dynamic Organization of Nuclear Function
  • Place of Presentation: N. Y. /USA
  • Year and Date: 20080917-21
• [Presentation] Nuclear protein import and its significance to cell function (2008)
  • Author(s): Yoneda, Y.
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
  • Year and Date: 20080916-17
• [Presentation] Regulation of importin alphal expression (2008)
  • Author(s): Yasunao Katnikawa, Noriko Yasiahara, Yoshihiro Yoneda
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
  • Year and Date: 20080916-17
• [Presentation] As novel regulatory system to maintain ES cells through nuclear-cytoplasmic transport (2008)
  • Author(s): Yasuhara, N, Memood, R., Kamikawa, Y., Touma, K. and Yoneda, Y
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
  • Year and Date: 20080916-17
• [Presentation] Nuclear-cytoplasmic transport regulation of transcription factors (2008)
  • Author(s): Memood, R., Yoshikawa, K., Shibazaki, N., Kaneko, A., Yasuhara, N., Kondoh, H. and Yoneda, Y
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
  • Year and Date: 20080916-17
• [Presentation] Nuclear import mechanism of Nanog (2008)
  • Author(s): Gibo, R., Okada, M., Yasuhara, and Yoneda, Y.
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
  • Year and Date: 20080916-17
• [Presentation] Selective localization of PCBP2 to P-bodies -An implication for functional diversity among P-hodies (2008)
  • Author(s): Ken Fujimura, Jun Katahira, Fumi Kano, Yoshihiro Yoneda, Masayuki Murata
  • Organizer: Cold Spring Harbor Laboratory Meeting on Dynamic Organization of Nuclear Function
  • Place of Presentation: N. Y. /USA
  • Year and Date: 20080903-07
• [Presentation] Nucleocytoplasmic transport of proteins and RNAs and its significance, to cell function (2008)
  • Author(s): Yoneda, Katahira, J., S ekimoto, T., Yasuhara, N., and Oka, M
  • Organizer: 第60回日本細胞生物学会大会
  • Place of Presentation: 横浜
  • Year and Date: 20080629-0701
• [Presentation] Functional differences between, two isoforms of Npap60(Nup50)in nuclear impiort (2008)
  • Author(s): Ogawa, Y., Oka, M., Asally, M. and Yoneda, Y.
  • Organizer: 第60回日本細胞生物学会大会
  • Place of Presentation: 横浜
  • Year and Date: 20080629-0701
• [Presentation] Nuclear accumulation of importin a suppresses DNA repliation (2008)
  • Author(s): Mizuguchi, C., Yasuda, Y. and Yoneda, Y.
  • Organizer: 第60回日本細胞生物学会大会
  • Place of Presentation: 横浜
  • Year and Date: 20080629-0701
• [Presentation] Analysis of potential roles of Nup98 FG repeat region in oncogenesis (2008)
  • Author(s): Masahiro Oka, Munehiro Asally, Yoshinori Yasuda, Yoshihiro Yoneda
  • Organizer: 第60回日本細胞生物学会大会
  • Place of Presentation: 横浜
  • Year and Date: 20080629-0701
• [Presentation] Identification of a cis-acting dendritic targeting element in the brain-derived neurotrophic factor mRNA (2008)
  • Author(s): Souiehi Oe, Yoshihiro Yoneda
  • Organizer: 第60回日本細胞生物学会大会
  • Place of Presentation: 横浜
  • Year and Date: 20080629-0701
• [Book] 分子生物学イラストレイテッド第3版 (2009)
  • Author(s): 浅利宗弘、米田悦啓
  • Total Pages: 348
  • Publisher: 羊土社
• [Book] 標準細胞生物学 (2008)
  • Author(s): 片平じゅん、米田税啓
  • Total Pages: 358
  • Publisher: 医学書院
• [Remarks]
  • URL: http://www.anat3.med.osaka-u.ac.ip/