核-細胞質間蛋白質輸送制御と細胞核ストレス応答

2008 Fiscal Year Annual Research Report

• Project/Area Number: 16107004
• Research Institution: Osaka University
• Principal Investigator: 米田 悦啓 Osaka University, 生命機能研究科, 教授 (80191667)
• Keywords: 核蛋白質輸送 / importin / Ran / ストレス応答 / p53 / 細胞核

Research Abstract

核-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やヌクレオポリンなど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解明された。しかし、核-細胞質問蛋白質輸送が、個体発生、細胞分化・増殖、細胞周期といった様々な生命現象とどのように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。一方、われわれは、これまでの研究から、importin αが単独で核内に移行する能力を有することを初めて発見するとともに、細胞が紫外線照射や熱ショック、酸化ストレスなどの様々なストレスを受けた時に、そのストレスに応答して速やかに核内に集積するという現象を見出してきた。本研究は、ストレスに応答して核内に集積したimportin αが、核内でどのような機能を発揮し、ストレスに対応するのかを知る目的で研究を進めた。本年度は、importin α核内集積条件下でDNAマイクロアレイ解析を行ない、発現が有意に減少する遺伝子に着目して研究を進めた。発現が減少する遺伝子は約60種類存在していたが、その中に、ATF3, Gadd45α, HRKなど、アポトーシスを誘導することが知られている遺伝子が複数含まれていることがわかった。これらの遺伝子発現はRT-PCRを用いた解析から、importin αによって制御を受けることが示され、importin αは、ストレスに応答して核に蓄積し、アポトーシスを回避するための転写制御に関与しているのではないかということが示唆された。また、importin αがDNAに直接結合することを見出していたが、p53と結合することにより、p53の機能を制御している可能性を検討し、レポーターアッセイを行った結果、p53による転写を負に制御していることが示唆された。

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 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 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 β for FG-nucleoporins 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
• [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ップ
  • Place of Presentation: 姫路
  • Year and Date: 20090126-20090128
• [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: 20081209-20081212
• [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, Davi Jans, Kate Loveland
  • 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: 20081209-20081212
• [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: 20080917-20080921
• [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: 20080917-20080921
• [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
  • Year and Date: 20080917-20080921
• [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 on Dynamic Organization of Nuclear Function
  • Place of Presentation: N. Y. /USA
  • Year and Date: 20080917-20080921
• [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: 20080917-20080921
• [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: 20080916-20080917
• [Presentation] Regulation of importin alphal expression (2008)
  • Author(s): Yasunao Kamikawa, Noriko Yasuhara, Yoshihiro Yoneda
  • Organizer: 2nd International SOX Meeting
  • Place of Presentation: 兵庫
  • Year and Date: 20080916-20080917
• [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
  • Year and Date: 20080916-20080917
• [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
  • Year and Date: 20080916-20080917
• [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
  • Year and Date: 20080916-20080917
• [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
  • Year and Date: 20080903-20080907
• [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
  • Year and Date: 20080629-20080701
• [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6O回 日本細胞生物学会大会
  • Place of Presentation: 横浜
  • Year and Date: 20080629-20080701
• [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
  • Year and Date: 20080629-20080701
• [Presentation] Analysis of potential roles of Nup98 FG repeat region in oncogenesis (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
  • Year and Date: 20080629-20080701
• [Presentation] Identification of a cis-acting 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
  • Year and Date: 20080629-20080701
• [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
• [Remarks]
  • URL: http://www.anat3.med.osaka-u.ac.jp/