Ranサイクルと細胞老化

2011 Fiscal Year Annual Research Report

• Project/Area Number: 23657130
• Research Institution: Osaka University
• Principal Investigator: 米田 悦啓 大阪大学, 生命機能研究科, 教授 (80191667)
• Project Period (FY): 2011-04-28 – 2012-03-31
• Keywords: 核輸送 / 細胞老化 / Ran / importin / RanBP1

Research Abstract

核-細胞質間分子輸送機構は、輸送因子であるimportinやexportin、輸送の方向性を決定する因子である低分子量GTPase Ran、輸送の通路である核膜孔複合体を構成する因子(ヌクレオポリン)の理解が進み、基本的な輸送メカニズムが提唱されている。一方、本提案者らはRanやRanBP1(=Ran binding protein 1)に関する研究を行っている過程で、正常細胞の細胞老化が進行するにつれて、Ranの発現量が徐々に低下するとともに、細胞増殖が盛んに行われている際には主として核に存在しているRanが、細胞老化に伴って徐々に細胞質に偏在していくことを発見した。これらの知見に基づき、Ranサイクルの変化とそれに伴う核-細胞質間分子輸送機構の変化が、細胞老化進行の根幹となっているのではないかと考え、Ranサイクルと細胞老化の関係を明らかにすることを目的として研究を進めた。MEF(=mouse embryonic fibroblast)細胞を用いて、継代培養を続けて行くと、確かにRanの発現量が低下し、その局在が細胞質側に偏移していくことが確認でき、その変化に応じて、核輸送因子であるimportin αの局在が変化するとともに、その発現量が低下して行くことが明らかになった。また、MEF細胞において、人為的にRanの発現量をノックダウンさせると、senescence-associated beta-galactosidase(SA-β-gal)活性が上昇し、細胞老化が進展することが明らかとなった。さらに、Ranのノックダウンを行ったMEF細胞において、MnSOD活性の上昇がみられることがわかった。以上より、細胞老化とRanが密接に関わっていることを明らかにすることができた。

Research Products

• [Journal Article] Cell type-dependent gene regulation by Staufen2 in conjunction with Upf1 (2011)
  • Author(s): Miki, T., Kurono, S., Kaneko, Y., Katahira, J. and Yoneda, Y.
  • Journal Title: BMC Mol. Biol Volume: 12 Pages: 48
  • DOI: doi:10.1186/1471-2199-12-48
  • Peer Reviewed
• [Journal Article] Stimulation of CK2-dependent Grp94 phosphorylation by the nuclear localization signal peptide (2011)
  • Author(s): Miyata, Y., Yoneda, Y. and Yahara, I.
  • Journal Title: Mol. Cell. Biochem Volume: 356 Pages: 191-200
  • DOI: DOI:10.1007/s11010-011-0944-9
  • Peer Reviewed
• [Journal Article] Nuclear import of serum response factor in airway smooth mascle (2011)
  • Author(s): McConville, J. F., Fernandes, D. J., Churchill, J., Dewundara, S., Kogut, P., Shah, S., Fuchs, G., Kedainis, D., Bellam, S. K., Patel, N. M., McCauley, J., Dulin, N. O., Gupta, M. P., Adam, S., Yoneda, Y., Camoretti-Mercado, B. and Solway, J.
  • Journal Title: Am J. Respir. Cell Mol. Biol Volume: 45 Pages: 453-458
  • DOI: doi:10.1165/rcmb.2008-0393OC
  • Peer Reviewed
• [Journal Article] NUP358, a nucleoporin, functions as a key determinant of the nuclear pore complex structure during skeletal myogensis (2011)
  • Author(s): Asally, M., et al
  • Journal Title: FEBS J. Volume: 585 Pages: 791-796
  • DOI: doi:10.1111/j.1742-4658.2010.07982.x
  • Peer Reviewed
• [Journal Article] Axotomy induces axonogenesis in hippocampal neurons by a mechanism dependent on importin β (2011)
  • Author(s): Ohara, R., et al
  • Journal Title: Biochem.Biophys.Res.Commun. Volume: 405 Pages: 697-702
  • DOI: 10.1016/j.bbrc.2011.01.108
  • Peer Reviewed
• [Journal Article] Mice lacking Ran Binding Protein 1 are viable and show male infertility (2011)
  • Author(s): Nagai, M., et al
  • Journal Title: FEBS Lett. Volume: 585 Pages: 791-796
  • DOI: 10.1016/j.bbrc.2011.01.108
  • Peer Reviewed
• [Journal Article] Proteomic analysis of importin α-interacting proteins in adult mouse brain (2011)
  • Author(s): Fukumoto, M., Sekimoto, T., Yoneda, Y.
  • Journal Title: Cell Struct.Funct. Volume: 36 Pages: 57-67
  • DOI: doi:10.1247/csf.10026
  • Peer Reviewed
• [Journal Article] Importin α protein acts as a negative regulator for Snail protein nuclear import (2011)
  • Author(s): Sekimoto, T., et al
  • Journal Title: J.Biol.Chem. Volume: 286 Pages: 15126-15131
  • DOI: doi:10.1074/jbc.M110.213579
  • Peer Reviewed
• [Journal Article] Generation of rat monoclonal antibody specific for mouse importin α8 (2011)
  • Author(s): Moriyama, T., Mizuguchi, C., Yoneda, Y.
  • Journal Title: Hybridoma Volume: 30 Pages: 303-306
  • DOI: doi:10.1089/hyb.2011.0004
  • Peer Reviewed
• [Journal Article] Generation and characterization of a monoclonal antibody against importin α7/NPI-2 (2011)
  • Author(s): Mizuguchi, C., Moriyama, T., Yoneda, Y.
  • Journal Title: Hybridoma Volume: 30 Pages: 307-309
  • DOI: doi:10.1089/hyb.2011.0006
  • Peer Reviewed
• [Journal Article] Targeted disruption of one of the importin α family members leads to female functional incompetence in delivery (2011)
  • Author(s): Moriyama, T., et al
  • Journal Title: FEBS J. Volume: 278 Pages: 1561-1572
  • DOI: doi:10.1111/j.1742-4658.2011.08079.x
  • Peer Reviewed
• [Journal Article] Cell type-specific transcriptional regulation of the gene encoding importin-α1 (2011)
  • Author(s): Kamikawa, Y., Yasuhara, N., Yoneda, Y.
  • Journal Title: Exp.Cell Res. Volume: 317 Pages: 1970-1978
  • DOI: 10.1016/j.yexcr.2011.05.024
  • Peer Reviewed
• [Journal Article] Cross talk between distinct nuclear import pathways enables efficient nuclear import of E47 in conjunction with its partner transcription factors (2011)
  • Author(s): Mehmood, R., et al
  • Journal Title: Mol.Biol.Cell Volume: 22 Pages: 3715-3724
  • DOI: doi:10.1091/mbc.E10-10-0809
  • Peer Reviewed
• [Journal Article] Nuclear retention of importin α coordinates cell fate through changes in gene expression (2011)
  • Author(s): Yasuda, Y., et al
  • Journal Title: EMBO J. Volume: 31 Pages: 83-94
  • DOI: doi:10.1038/emboj.2011.360
  • Peer Reviewed
• [Journal Article] Nucleo-cytoplasmic transport of micrornas and related small RNAs (2011)
  • Author(s): Katahira, J., Yoneda, Y.
  • Journal Title: Traffic Volume: 12 Pages: 1468-1474
  • DOI: doi:10.1111/j.1600-0854.2011.01211.x
  • Peer Reviewed
• [Journal Article] The role of Rael in Nup98-fusion mediated leukemogenesis (2011)
  • Author(s): Oka, M., Yoneda, Y.
  • Journal Title: Cell Cycle. Volume: 10 Pages: 12
  • DOI: 10.4161/cc.10.12.15759
  • Peer Reviewed
• [Journal Article] The interaction between importin-alpha and Nup153 promotes importin-alpha/beta-mediated nuclear import (2011)
  • Author(s): Ogawa, Y., et al
  • Journal Title: Traffic Volume: (In press)
  • Peer Reviewed
• [Journal Article] Nuclear importin α and its physiological importance (2011)
  • Author(s): Miyamoto, Y., et al
  • Journal Title: Commun.Integ.Biol. Volume: (In press)
  • Peer Reviewed
• [Journal Article] Intrinsic and extrinsic negative regulators of nuclear protein transport processes (2011)
  • Author(s): Sekimoto, T., Yoneda Y.
  • Journal Title: Genes Cells Volume: (In press)
  • Peer Reviewed
• [Journal Article] The small GTPase Ran and Ran-binding proteins (2011)
  • Author(s): Nagai, M., Yoneda, Y.
  • Journal Title: BioMol.Concepts Volume: (In press)
  • Peer Reviewed
• [Presentation] The involvement of small GTPase Ran in cellular aging (2011)
  • Author(s): 永井理博、米田悦啓
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)(招待講演)
  • Year and Date: 20111213-20111216
• [Presentation] Importin alphamediates biased translocation through nuclear pores in conjunction withNup153 (2011)
  • Author(s): Ogawa, Y., et al
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 20111213-20111216
• [Presentation] The mechanism to determine a nuclear transport pathway by NLS sequence (2011)
  • Author(s): Kimoto, C., Yasuhara, N. and Yoneda, Y.
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 20111213-16
• [Presentation] Role of nucleocytoplasmic shuttling of estrogen receptor (2011)
  • Author(s): Moriyama, T., Oka, M. and Yoneda, Y.
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 20111213-16
• [Presentation] The nucleocytoplasmic transport system regulates the stem cell differentiation (2011)
  • Author(s): Yasuhara, N., Yamagishi, R., Arai, Y., Yanagida, T., Kaneko, H. and Yoneda, Y.
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 20111213-16