核-細胞質間物質輸送制御の視点からの脳性差発現機構解明へのアプローチ

• Project/Area Number: 19657062
• Research Category: Grant-in-Aid for Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Cell biology
• Research Institution: Osaka University
• Principal Investigator: 米田 悦啓 Osaka University, 生命機能研究科, 教授 (80191667)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2008: ¥1,700,000 (Direct Cost: ¥1,700,000); Fiscal Year 2007: ¥1,900,000 (Direct Cost: ¥1,900,000)
• Keywords: 核蛋白質輸送 / importin α / importin β / 性差 / 脳機能 / 核輸送因子 / 質量分析法 / ノックアウトマウス

Research Abstract

核一細胞質間蛋白質輸送機構に関する研究は、importin βファミリー分子、importin αファミリー分子、低分子量GTPase Ranやそのサイクルに関する因子群の性状解析が精力的に進められ、輸送機構に関する基本的な分子メカニズムが理解される段階にまで研究は進展した。しかし、核一細胞質間蛋白質輸送が、個体発生、細胞分化・増殖、細胞死、ストレス応答、細胞周期といった様々な生命現象とどのように深く関連しているのかという点についてはほとんど研究が進んでいない。本研究では、マウス脳において蛋白質の核輸送機構に性差による制御が存在し、それらが脳の性差発現に重要であるのではないかという仮説の下に、1)性差に着目したマウス脳における核輸送因子発現の網羅的解析、2)発現に性差が見られる核輸送因子により運ばれている脳の性差決定(維持)候補因子の同定、3)性差の見られた核輸送因子の脳内特定部位特異的なコンディショナルノックアウトマウス、及び脳の性差決定(維持)候補因子のノックアウトマウスを用いた脳性差発現の解析を行うことを目的として研究を進めた。昨年度の研究の結果、importin αファミリーに属する分子の1つであるimportin α5(NPI-1)の発現量が、蛋白質のレベルで、雌の脳(小脳、中脳、前頭葉、海馬、嗅球など)において増加していることが明らかとなった。一方、importin βの発現量には顕著な雌雄差は見られなかった。そこで、本年度は、神経細胞においてimportin α5によって輸送されている因子群の、質量分析法による網羅的同定を試みた。その結果、計47種類の脳の性差を決定する可能性のある候補分子を同定することができた。

Research Products

• [Journal Article] Infrared laser-mediated gene induction in targeted single cells in vivo. (2009)
  • Author(s): Kamei, Y., et al.
  • Journal Title: Nature Methods 6(in press) Pages: 79-81
  • Peer Reviewed
• [Journal Article] mDia2 shuttles between the nucleus and the cytoplasm through the Importin-α/β- and CRM1-mediated nuclear transport mechanism. (2009)
  • Author(s): Miki, T., et al.
  • Journal Title: J. Biol. Chem. 284(in press) 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 NeuroDl 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., 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: Neuron 59 Pages: 241-252
  • Peer Reviewed
• [Journal Article] Individual binding pockets of importin βfor FG-nuclepporins are differently regulated by RanGTP. (2008)
  • Author(s): Otsuka, S., et al.
  • Journal Title: Proc. Natl Acad. Sci. USA 105 Pages: 16101-16106
  • Peer Reviewed
• [Journal Article] Characterization of human herpesvirus 6 U69 gene product and identification of its nuclear localization signal. (2008)
  • Author(s): Isegawa, Y., et. al.
  • Journal Title: J. Virol. 82 Pages: 710-718
  • Peer Reviewed
• [Journal Article] Nuclear RNA export factor 7 is localized in processing bodies and neuronal mRNA granules through interactions with shuttling hnRNPs. (2008)
  • Author(s): Katahira, J., et. al.
  • Journal Title: Nucl. Acids Res. 36 Pages: 616-628
  • Peer Reviewed
• [Journal Article] NXF2 is involved in cytoplasmic mRNA dynamics through interactions with motor proteins. (2007)
  • Author(s): Takano, K., et. al.
  • Journal Title: Nucl. Acids Res. 35 Pages: 2513-2521
  • Peer Reviewed
• [Journal Article] A novel nuclear import of Vpr promoted by importin α is crucial for HIV-1 replication in macrophages. (2007)
  • Author(s): Nitahara-Kasahara, Y., et. al.
  • Journal Title: J. Virol. 81 Pages: 5284-5293
  • Peer Reviewed
• [Journal Article] Arxl functions as an unorthodox nuclear export receptor for the 60Spre-ribosomal subunit. (2007)
  • Author(s): Bradatsch, B., et. al.
  • Journal Title: Mol. Cell 27 Pages: 767-779
  • Peer Reviewed
• [Journal Article] Normal brain development in importin-α5 deficient-mice. (2007)
  • Author(s): Yasuhara, N., et. al.
  • Journal Title: Nature Cell Biol. 9 Pages: 1339-1339
  • Peer Reviewed
• [Journal Article] 細胞分化を制御する核輸送因子importin α (2007)
  • Author(s): 安原徳子、米田悦啓
  • Journal Title: 蛋白質核酸酵素 52 Pages: 427-433
  • NAID: 40015447931
  • 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., Yoneda, Y.
  • Organizer: 第26回染色体ワークショツプ
  • Place of Presentation: 姫路
• [Presentation] Nucleocytoplasmic transport of proteins and RNAs and its significance to cell function (2008)
  • Author(s): Yoneda, Y., Katahira, J., Sekimoto, T., Yasuhara, N., Oka, M.
  • Organizer: 第60回 日本細胞生物学会大会
  • Place of Presentation: 横浜
• [Presentation] Functional differences between two isoforms of Npap60 (Nup50) in nuclear impiort (2008)
  • Author(s): Ogawa, Y., Miyamoto, Y., Oka, M., Asally, M., Yoneda, Y.
  • Organizer: 第60回 日本細胞生物学会大会
  • Place of Presentation: 横浜
• [Presentation] Nuclear accumulation of importin a suppresses DNA replication (2008)
  • Author(s): Mizuguchi, C, Yasuda, Y., Yoneda, Y.
  • Organizer: 第60回 日本細胞生物学会大会
  • Place of Presentation: 横浜
• [Presentation] Analysis of potential roles of Nup98 FG repeat region inoncogenesis (2008)
  • Author(s): Masahiro Oka, Munehiro Asally, Yoshinari Yasuda, Yoshihiro Yoneda
  • Organizer: 第60回 日本細胞生物学会大会
  • Place of Presentation: 横浜
• [Presentation] Identification of a cis-aeting dendritic targeting element in the brain-derived neurotrophic factor mRNA. (2008)
  • Author(s): Souichi Oe, Yoshihiro Yoneda
  • Organizer: 第60回 日本細胞生物学会大会
  • Place of Presentation: 横浜
• [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
• [Presentation] Regulation of importin alpha 1 expression (2008)
  • Author(s): Yasunao Kamikawa, NorikoYasuhara, Yoshihiro Yoneda
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
• [Presentation] A novel regulatory system to maintain ES cells through nuclear-cytoplasmic transport (2008)
  • Author(s): Yasuhara, N, Memood, R., Kamikawa, Y., Touma, K., Yoneda, Y.
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
• [Presentation] Nuclear-cytoplasmic transport regulation of transcription factors (2008)
  • Author(s): Memood, R., Yoshikawa, K. Shibazaki, N., Kaneko, A. Yasuhara, N., Kondoh, H., Yoneda, Y.
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
• [Presentation] Nuclear import mechanism of Nanog (2008)
  • Author(s): Gibo, R., Okada, M., Yasuhara, N., Yoneda, Y.
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
• [Presentation] Selective localization of PCBP2 to P-bodies - An implicationfor functional diversity among P-bodies (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
• [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
• [Presentation] Nuclear import mechanism of neuroDl, 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
• [Presentation] The role of Nup98 in a Crm1-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
• [Presentation] Functional analysis of nuclear importin α accumulating in response to the stress. (2008)
  • Author(s): Yasuda, Y., Miyamoto, Y., Yoneda, Y.
  • Organizer: Cold Spring Harbor Laboratory Meeting onDynamic Organization of Nuclear Function
  • Place of Presentation: N. Y. /USA
• [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
• [Presentation] 核蛋白質輸送と生命機能 (2008)
  • Author(s): 米田悦啓
  • Organizer: BMB2008 第3 1回日本分子生物学会年会第8 1回日本生化学会大会合同大会
  • Place of Presentation: 神戸
• [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3 1回日本分子生物学会年会・第8 1回日本生化学会大会合同大会
  • Place of Presentation: 神戸
• [Presentation] 核-細胞質間分子流通機構と生命機能の調節 (2007)
  • Author(s): 米田悦啓
  • Organizer: 第71回日本生化学会中部支部会
  • Place of Presentation: 名古屋
  • Year and Date: 2007-05-19
• [Presentation] Importin alpha has multiple functions (2007)
  • Author(s): Yoneda, Y.
  • Organizer: EMBO Workshop on Mechanisms of Nucleocytoplasmic Transport
  • Place of Presentation: Italy
• [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, 米田悦啓
  • Total Pages: 358
  • Publisher: 医学書院
• [Book] Nuclear Dynamics (2007)
  • Author(s): Saiwaki, T. and Yoneda, Y.
  • Total Pages: 279
  • Publisher: Springer-Verlag Inc.
• [Book] 細胞核の世界-ダイナミクスから病態まで (2007)
  • Author(s): 米田悦啓、他
  • Total Pages: 383
  • Publisher: 共立出版
• [Remarks]
  • URL: http://www.anat3.med.osaka-u.ac.jp/
• [Remarks]
  • URL: http://www.anat3.med.osaka-u.ac.jp/