核－細胞質間の物質輸送を制御する核膜孔複合体内の分子動態の解明

• Project Area: Spying minority in biological phenomena -Toward bridging dynamics between individual and ensemble processes-
• Project/Area Number: 24115512
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyoto University
• Principal Investigator: 粂田 昌宏 京都大学, 生命科学研究科, 助教 (00582181)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000); Fiscal Year 2013: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2012: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
• Keywords: 核膜孔複合体 / 核輸送 / 輸送タンパク質 / 分子個性 / インポーチン

Research Abstract

核膜孔複合体（NPC）を介した分子輸送の基本的メカニズムを理解するため、昨年までの「通過分子の構造変化」に着目した解析に加え、細胞あたり4000個ほど存在する「NPCごとの機能的個性」に着目した解析を開始した。実験系としては、全反射型顕微鏡を斜方照明にして用いた一分子蛍光測定系を立ち上げ、GFP融合タンパク質がどのように異なるNPCを通過して核内に到達するかを経時観察し、結果をNPCごとに分けて解析した。その結果、分子の通過頻度と滞在時間はともに、NPCごとに4-5倍程度の大きなばらつきがあることが分かった。このことは、NPCは機能的にヘテロであり、様々な性質のNPCが一細胞核に共存することで適時適切な核輸送が達成されていることを意味するものである。現在、NPC内部でのサブユニット間のSS結合に着目して、このNPCごとに個性が現れる分子基盤を解明すべく取り組んでいる。これらの研究は、生体高分子にみられる分子個性の典型的な例として、今後のこの分野の研究にとって重要な知見であると考えている。

Current Status of Research Progress

Reason

25年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

25年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Probing the stiffness of isolated nucleoli by atomic force microscopy. (2014)
  • Author(s): Louvet E, Yoshida A, Kumeta M, Takeyasu K.
  • Journal Title: Histochemistry and Cell Biology Volume: 141 Issue: 4 Pages: 365-81
  • DOI: 10.1007/s00418-013-1167-9
  • Peer Reviewed
• [Journal Article] Caprice/MISP is a novel F-actin bundling protein critical for actin-based cytoskeletal reorganizations. (2014)
  • Author(s): Kumeta M., Gilmore J. L., Umeshima H., Ishikawa M., Kitajiri S., Horigome T., Kengaku M., and Takeyasu K
  • Journal Title: Genes Cells Volume: 19 Issue: 4 Pages: 338-349
  • DOI: 10.1111/gtc.12131
  • Peer Reviewed / Open Access
• [Journal Article] Dynamics of WD-repeat containing proteins in SSU processome components (2014)
  • Author(s): Wada K, Sato M, Araki N, Kumeta M, Hirai Y, Takeyasu K, Furukawa K, Horigome T.
  • Journal Title: Biochemistry and Cell Biology Volume: -
  • Peer Reviewed
• [Journal Article] Intermolecular disulfide bonds among nucleoporins regulate karyopherin-dependent nuclear transport (2013)
  • Author(s): Yoshimura SH, Otsuka S, Kumeta M, Taga M, Takeyasu K
  • Journal Title: J. Cell Sci. Volume: 126 Pages: 3141-3150
  • DOI: 10.1242/jcs.124172
  • Peer Reviewed
• [Journal Article] Antibody-based analysis reveals "filamentous vs. non-filamentous" and "cytoplasmic vs. nuclear" crosstalk of cytoskeletal proteins (2013)
  • Author(s): Kumeta M, Hirai Y, Yoshimura SH, Horigome T, Takeyasu K
  • Journal Title: Exp. Cell Res. Volume: 319 Issue: 20 Pages: 3226-3237
  • DOI: 10.1016/j.yexcr.2013.07.021
  • NAID: 120005353499
  • Peer Reviewed
• [Journal Article] Nucleolar scaffold protein, WDR46, determines the granular compartmental localization of nucleolin and DDX21 (2013)
  • Author(s): Hirai Y, Louvet E, Oda T, Kumeta M, Watanabe Y, Horigome T, Takeyasu K
  • Journal Title: Genes Cells Volume: 18 Issue: 9 Pages: 780-797
  • DOI: 10.1111/gtc.12077
  • Peer Reviewed
• [Journal Article] Interaction, mobility, and phosphorylation of human orthologues of WD repeat-containing components of the yeast SSU processome t-UTP sub-complex. (2013)
  • Author(s): Sato M, Araki N, Kumeta M, Takeyasu K, Taguchi Y, Asai T, Furukawa K, Horigome T.
  • Journal Title: Biochemistry and Cell Biology Volume: 91 Issue: 6 Pages: 466-75
  • DOI: 10.1139/bcb-2013-0062
  • Peer Reviewed
• [Journal Article] AFM investigation of the organization of actin bundles formed by actin-binding proteins. (2013)
  • Author(s): Gilmore J, Kumeta M, Takeyasu K.
  • Journal Title: Journal of Surface Engineered Materials and Advanced Technology Volume: 3 Pages: 13-19
  • Peer Reviewed
• [Journal Article] Karyopherin-independent spontaneous transport of amphiphilic proteins through the nuclear pore (2012)
  • Author(s): Kumeta M
  • Journal Title: J. Cell Sci Volume: 125 Pages: 4979-84
  • DOI: 10.1242/jcs.109520
  • Peer Reviewed
• [Presentation] Spontaneous transport of amphiphilic proteins through the nuclear pore (2014)
  • Author(s): Kumeta M, Lolodi O, Yoshimura SH, Takeyasu K.
  • Organizer: 58th Annual Meeting of Biophysical Society
  • Place of Presentation: San Francisco, USA
• [Presentation] 核－細胞質間の物質輸送を制御する核膜孔複合体内の分子動態 (2013)
  • Author(s): 粂田昌宏
  • Organizer: 第65回日本細胞生物学会大会
  • Place of Presentation: 名古屋
• [Presentation] Analysis of nuclear transport of single molecules by narrow-field illumination microscopy (2013)
  • Author(s): Lolodi O, Fujiwara T, Takeyasu K, Kumeta M.
  • Organizer: The 10th MicRO Alliance Meeting and Nanomics Symposium
  • Place of Presentation: 京都
• [Presentation] Nucleocytoplasmic transport of macromolecules: the nature of the hydrophobic barrier inside the nuclear pore (2012)
  • Author(s): Kumeta M
  • Organizer: 2012 Paradigm Innovation in Biology: Novel Strategy and Thinking
  • Place of Presentation: Academia Sinica Taipei, Taiwan
  • Year and Date: 2012-10-16