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Functional Dynamics of Liquid-Liquid Phase Separation Mediated by Nuclear Pore Complex

Research Project

Project/Area Number 21K19043
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 37:Biomolecular chemistry and related fields
Research InstitutionKanazawa University

Principal Investigator

Wong Richard  金沢大学, ナノ生命科学研究所, 教授 (30464035)

Co-Investigator(Kenkyū-buntansha) 小林 亜紀子  金沢大学, 新学術創成研究機構, 特任助教 (00345662)
羽澤 勝治  金沢大学, 新学術創成研究機構, 准教授 (40622460)
LIM KEE・SIANG  金沢大学, ナノ生命科学研究所, 特任助教 (60842987)
Project Period (FY) 2021-07-09 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2022: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2021: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords核膜孔複合体 / 核膜孔 / 液-液相分離 / 翻訳後修飾 / 相分離
Outline of Research at the Start

本研究は、高次遺伝子発現システムの基盤となる核膜孔複合体(Nuclear Pore Complex; NPC)の作動原理の解明にむけて、NPCが確立する液-液相分離(Liquid-Liquid Phase Separation; LLPS)の状態・機能相関を明らかにする。LLPSとは2つの液体が混ざり合わずに、互いに排除しあうことにより2相に分離する現象のことである。細胞核の働きについて新しい概念を創出し、広範な生命機能現象と多くの疾病症メカニズムに対する根本的な理解を目指すものである。このように、本研究課題は、新規NPC-LLPS研究領域の展開に挑戦するものである。

Outline of Final Research Achievements

In this study, we successfully tracked the nanoscale dynamics of the internal structure's biofilaments in the nuclear pore complex (NPC) using HS-AFM. We discovered that in colorectal cancer cells, unlike normal cells, the thickness of individual biofilaments becomes heterogeneous, leading to increased rotation and movement of biofilaments and active formation of cancer-specific plug structures. Additionally, through gene interference experiments, we revealed that specific FG-NUPs overexpressed in colorectal cancer cells impact the dynamics of biofilaments. As a result, we achieved the first visualization of the spatiotemporal behavior of biofilaments from their formation to disappearance within the LLPS environment of the NPC.

Academic Significance and Societal Importance of the Research Achievements

この研究は、数十年にわたって信じられてきた逆の結果を示し、NPC内のバイオフィラメントを追跡・操作することが可能になりました。この結果、がん細胞固有のバイオフィラメントの異常な動態と機能の相関を明らかにしました。これらの知見は、細胞内で様々な分子反応が起こるLLPS環境の物理的特性解析におけるHS-AFMの有用性を示すだけでなく、分子レベルでの核膜ダイナミクスの理解と制御に基づく新たながんの診断と治療法の開発に期待が持てます。

Report

(3 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • Research Products

    (51 results)

All 2023 2022 2021 Other

All Journal Article (13 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 13 results,  Open Access: 13 results) Presentation (31 results) (of which Int'l Joint Research: 1 results,  Invited: 7 results) Remarks (5 results) Patent(Industrial Property Rights) (2 results)

  • [Journal Article] Nanoscopic Assessment of Anti-SARS-CoV-2 Spike Neutralizing Antibody Using High-Speed AFM.2023

    • Author(s)
      Keesiang Lim, Goro Nishide, Elma Sakinatus Sajidah, Tomoyoshi Yamano, Yujia Qiu, Takeshi Yoshida, Akiko Kobayashi, Masaharu Hazawa, Toshio Ando, Rikinari Hanayama, Richard W Wong
    • Journal Title

      Nano letters

      Volume: 23 Issue: 2 Pages: 619-628

    • DOI

      10.1021/acs.nanolett.2c04270

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Spatiotemporal tracking of small extracellular vesicle nanotopology in response to physicochemical stresses revealed by HS-AFM.2022

    • Author(s)
      Elma Sakinatus Sajidah, Keesiang Lim, Tomoyoshi Yamano, Goro Nishide, Yujia Qiu, Takeshi Yoshida, Hanbo Wang, Akiko Kobayashi, Masaharu Hazawa, Firli R P Dewi, Rikinari Hanayama, Toshio Ando, Richard W Wong
    • Journal Title

      Journal of extracellular vesicles

      Volume: 11 Issue: 11 Pages: 12275-12275

    • DOI

      10.1002/jev2.12275

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Inhibition of Canonical Wnt Signaling Promotes Ex Vivo Maintenance and Proliferation of Hematopoietic Stem Cells in Zebrafish2022

    • Author(s)
      Kimura Koki、Yamamori Shiori、Hazawa Masaharu、Kobayashi-Sun Jingjing、Kondo Mao、Wong Richard W、Kobayashi Isao
    • Journal Title

      Stem Cells

      Volume: 40 Issue: 9 Pages: 831-842

    • DOI

      10.1093/stmcls/sxac044

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Discovery of a Novel Aminocyclopropenone Compound That Inhibits BRD4-Driven Nucleoporin NUP210 Expression and Attenuates Colorectal Cancer Growth2022

    • Author(s)
      Kondo Hiroya、Mishiro Kenji、Iwashima Yuki、Qiu Yujia、Kobayashi Akiko、Lim Keesiang、Domoto Takahiro、Minamoto Toshinari、Ogawa Kazuma、Kunishima Munetaka、Hazawa Masaharu、Wong Richard W.
    • Journal Title

      Cells

      Volume: 11 Issue: 3 Pages: 317-317

    • DOI

      10.3390/cells11030317

    • Related Report
      2022 Annual Research Report 2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] NSP9 of SARS-CoV-2 attenuates nuclear transport by hampering nucleoporin 62 dynamics and functions in host cells2022

    • Author(s)
      Makiyama Kei、Hazawa Masaharu、Kobayashi Akiko、Lim Keesiang、Voon Dominic C.、Wong Richard W.
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 586 Pages: 137-142

    • DOI

      10.1016/j.bbrc.2021.11.046

    • Related Report
      2022 Annual Research Report 2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Millisecond dynamic of SARS‐CoV‐2 spike and its interaction with ACE2 receptor and small extracellular vesicles2021

    • Author(s)
      Lim Keesiang、Nishide Goro、Yoshida Takeshi、Watanabe‐Nakayama Takahiro、Kobayashi Akiko、Hazawa Masaharu、Hanayama Rikinari、Ando Toshio、Wong Richard W.
    • Journal Title

      Journal of Extracellular Vesicles

      Volume: 10 Issue: 14

    • DOI

      10.1002/jev2.12170

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Synthetic zwitterions as efficient non-permeable cryoprotectants2021

    • Author(s)
      Kato Yui、Uto Takuya、Tanaka Daisuke、Ishibashi Kojiro、Kobayashi Akiko、Hazawa Masaharu、Wong Richard W.、Ninomiya Kazuaki、Takahashi Kenji、Hirata Eishu、Kuroda Kosuke
    • Journal Title

      Communications Chemistry

      Volume: 4 Issue: 1 Pages: 151-151

    • DOI

      10.1038/s42004-021-00588-x

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] New Activities of the Nuclear Pore Complexes2021

    • Author(s)
      Wong Richard W.
    • Journal Title

      Cells

      Volume: 10 Issue: 8 Pages: 2123-2123

    • DOI

      10.3390/cells10082123

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A light-switching pyrene probe to detect phase-separated biomolecules2021

    • Author(s)
      Hazawa Masaharu、Amemori Shogo、Nishiyama Yoshio、Iga Yoshihiro、Iwashima Yuki、Kobayashi Akiko、Nagatani Hirohisa、Mizuno Motohiro、Takahashi Kenji、Wong Richard W.
    • Journal Title

      iScience

      Volume: 24 Issue: 8 Pages: 102865-102865

    • DOI

      10.1016/j.isci.2021.102865

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Label-free tomographic imaging of nanodiamonds in living cells2021

    • Author(s)
      Ikliptikawati Dini Kurnia、Hazawa Masaharu、So Frederick T.-K.、Terada Daiki、Kobayashi Akiko、Segawa Takuya F.、Shirakawa Masahiro、Wong Richard W.
    • Journal Title

      Diamond and Related Materials

      Volume: 118 Pages: 108517-108517

    • DOI

      10.1016/j.diamond.2021.108517

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] How SARS-CoV-2 and Other Viruses Build an Invasion Route to Hijack the Host Nucleocytoplasmic Trafficking System2021

    • Author(s)
      Sajidah Elma Sakinatus、Lim Keesiang、Wong Richard W.
    • Journal Title

      Cells

      Volume: 10 Issue: 6 Pages: 1424-1424

    • DOI

      10.3390/cells10061424

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] High-Speed Atomic Force Microscopy Reveals Spatiotemporal Dynamics of Histone Protein H2A Involution by DNA Inchworming2021

    • Author(s)
      Nishide Goro、Lim Keesiang、Mohamed Mahmoud Shaaban、Kobayashi Akiko、Hazawa Masaharu、Watanabe-Nakayama Takahiro、Kodera Noriyuki、Ando Toshio、Wong Richard W.
    • Journal Title

      The Journal of Physical Chemistry Letters

      Volume: 12 Issue: 15 Pages: 3837-3846

    • DOI

      10.1021/acs.jpclett.1c00697

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] CDK8 maintains stemness and tumorigenicity of glioma stem cells by regulating the c-MYC pathway2021

    • Author(s)
      Fukasawa K, Kadota T, Horie T, Tokumura K, Terada R, Kitaguchi Y, Park G, Ochiai S, Iwahashi S, Okayama Y, Hiraiwa M, Yamada T, Iezaki T, Kaneda K, Yamamoto M, Kitao T, Shirahase H, Hazawa M, Wong RW, Todo T, Hirao A, Hinoi E.
    • Journal Title

      Oncogene

      Volume: ー Issue: 15 Pages: 2803-2815

    • DOI

      10.1038/s41388-021-01745-1

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Nano assessment of how viruses seize host nuclear transport systems2022

    • Author(s)
      Richard Wong
    • Organizer
      新学術創成研究機構ワークショップ
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Nano assessment of how viruses hijack host nuclear transport systems2022

    • Author(s)
      Richard Wong
    • Organizer
      15th NanoLSI Colloquium
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Nano imaging of genomic modality in nuclear pore territories2022

    • Author(s)
      Richard Wong
    • Organizer
      学術変革領域A 2022年度 ゲノムモダリティ領域会議
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] Nano imaging of nuclear pore dynamics in cancer cells2022

    • Author(s)
      Richard Wong
    • Organizer
      第95回日本生化学会大会シンポジウム
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] クロススケール核膜孔内選択的分子輸送システムの解明とその破綻2022

    • Author(s)
      Richard Wong
    • Organizer
      学術変革領域A クロススケール新生物学 第2回領域会議
    • Related Report
      2022 Annual Research Report
    • Invited
  • [Presentation] 核膜孔テリトリーにおける動的ナノ構造と機能2021

    • Author(s)
      Richard Wong
    • Organizer
      第73回日本細胞生物学会シンポジウム
    • Related Report
      2021 Research-status Report
    • Invited
  • [Presentation] ウイルスの核膜孔への侵入経路を解明2021

    • Author(s)
      Richard Wong
    • Organizer
      第6回新学術創成研究機構シンポジウム
    • Related Report
      2021 Research-status Report
    • Invited
  • [Presentation] The epigenetic role of nucleoporin RAE1 in human breast cancer2021

    • Author(s)
      Dini Kurnia Ikliptikawati, Akiko Kobayashi, Masaharu Hazawa, Richard W.Wong
    • Organizer
      The 39th Sapporo International Cancer Symposium
    • Related Report
      2021 Research-status Report
    • Int'l Joint Research
  • [Presentation] 核内相分離の制御を可能とする新規アミノシクロプロペノン化合物の発見2021

    • Author(s)
      近藤 寛弥、三代憲司、岩嶋友紀、Qiu Yujia、稲葉有香、小林亜紀子、Keesiang Lim、源利成、小川数馬、國嶋崇隆、羽澤勝治、Richard Wong
    • Organizer
      第39回染色体ワークショップ・第20回核ダイナミクス研究会
    • Related Report
      2021 Research-status Report
  • [Presentation] ヒストンH2A-DNAダイナミクスの実時空間イメージング2021

    • Author(s)
      西出梧朗、Keesiang Lim、小林亜紀子、羽澤勝治、稲葉有香、中山隆宏、古寺哲幸、安藤敏夫、Richard Wong
    • Organizer
      第39回染色体ワークショップ・第20回核ダイナミクス研究会
    • Related Report
      2021 Research-status Report
  • [Presentation] 核膜孔複合体による転写制御機構2021

    • Author(s)
      岩嶋友紀、羽澤勝治、Qiu Yujia、Dini Kurnia Ikliptikawati1、西 出 梧朗、小林 亜紀子、Lim Kee Siang、Richard Wong
    • Organizer
      第39回染色体ワークショップ・第20回核ダイナミクス研究会
    • Related Report
      2021 Research-status Report
  • [Presentation] 核膜孔複合体による核膜近傍での転写制御機構2021

    • Author(s)
      岩嶋友紀、羽澤勝治、Qiu Yujia、Dini Kurnia Ikliptikawati、西出 梧朗、小林 亜紀子、Lim Kee Siang、Richard Wong
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] 上衣腫における核膜孔複合体タンパク質Tprの役割2021

    • Author(s)
      小林亜紀子、Shabierjiang Jiapaer、Firli R.P. Dewi、羽澤勝治、Dini K. Ikliptikawati、Hartono、Hemragul Sabit、中田光俊、Richard Wong
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] SARS-CoV-2由来タンパク質NSP9の宿主細胞における核膜輸送に及ぼす影響の解明2021

    • Author(s)
      牧山 桂、羽澤 勝治、Richard W. Wong
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] SARS-CoV-2由来ORF6は核膜孔複合体RAE1とNUP98の局在変化を引き起こす2021

    • Author(s)
      加藤弘毅、小林亜紀子、羽澤勝治、Richard W Wong
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] Optical Diffraction Tomography as Label-Free High-Resolution 3-D Imaging of Nanodiamonds in Cancer Cell2021

    • Author(s)
      Dini Ikliptikawati, Masaharu Hazawa, Frederick So, Daiki Terada, Akiko Kobayahi, Takuya Segawa, Masahiro Shirakawa, Richard Wong
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] Molecular Dynamic of Coronavirus Spike Protein2021

    • Author(s)
      Keesiang Lim, Goro Nishide, Takeshi Yoshida, Takahiro Watanabe-Nakayama, Akiko Kobayashi, Masaharu Hazawa, Rikinari Hanayama, Toshio Ando, Richard W. Wong
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] 液-液相分離の動態解析を可能とする機能性蛍光プローブ2021

    • Author(s)
      羽澤 勝治、雨森 翔悟、西山 嘉男、岩嶋 友紀、小林 亜紀子、永谷 広久、水野 元博、髙橋 憲司、ウォング リチャード
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] Investigation of real-time dynamic histone H2A-DNA interaction and H2A-DNA condensation/de-condensation using high-speed AFM2021

    • Author(s)
      西出梧朗、Keesiang LIM、小林亜紀子、羽澤勝治、中山隆宏、古寺哲幸、安藤敏夫、Richard WONG
    • Organizer
      第59回日本生物物理学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] 高速AFMを用いたヒストンH2A-DNA実時空間イメージングによるヌクレオソーム凝縮プロセスとその環境の解明2021

    • Author(s)
      西出梧朗、Keesiang Lim、Mahmoud Shaaban Mohamed、小林亜紀子、羽澤勝治、中山隆宏、古寺哲幸、安藤敏夫、Richard Wong
    • Organizer
      第94回日本生化学大会
    • Related Report
      2021 Research-status Report
  • [Presentation] 新規アミノシクロプロぺノン化合物はBRD4依存的なNUP210の発現を阻害し、がん細胞の増殖を抑制する2021

    • Author(s)
      近藤 寛弥、三代憲司、羽澤勝治、岩嶋友紀、小林亜紀子、小川数馬、Richard Wong
    • Organizer
      第94回日本生化学会大会
    • Related Report
      2021 Research-status Report
  • [Presentation] NUP153 drives oncogenic TP63 expression through liquid-liquid phase separation mediated gene-gating in squamous cancer2021

    • Author(s)
      Masaharu Hazawa, Takeshi Suzuki, Toshinari Minamoto, Richard Wong
    • Organizer
      第80回 日本癌学会学術総会
    • Related Report
      2021 Research-status Report
  • [Presentation] 高速 AFM によるヒストン H2A トポグラフィーとその DNA コンパクション時の時空間ダイナミクスの解明2021

    • Author(s)
      西出梧朗、Keesiang Lim、小林亜紀子、羽澤勝治、中山隆宏、古寺哲幸、安藤敏夫、Richard Wong
    • Organizer
      第21回日本蛋白質科学会年会
    • Related Report
      2021 Research-status Report
  • [Presentation] Karyopherin-b1 regulates radioresistance in head and neck squamous cell carcinoma cell lines2021

    • Author(s)
      Masaharu Hazawa, Richard Wong, Ikuo Kashiwakura, Hironori Yoshino
    • Organizer
      日本放射線影響学会第64回大会
    • Related Report
      2021 Research-status Report
  • [Presentation] 核膜孔によるスーパーエンハンサーの局在・機能制御機序の解明2021

    • Author(s)
      羽澤 勝治、Richard Wong
    • Organizer
      第73回日本細胞生物学会大会
    • Related Report
      2021 Research-status Report
  • [Presentation] 高速 AFM を用いた DNA インチワーミングによるヒストン H2A ラッピング動態の実時空間イメージング2021

    • Author(s)
      西出梧朗、Keesiang Lim、小林亜紀子、羽澤勝治、中山隆宏、古寺哲幸、安藤敏夫、Richard Wong
    • Organizer
      第73回日本細胞生物学会大会
    • Related Report
      2021 Research-status Report
  • [Presentation] 新規アミノシクロプロペノン化合物は BRD4 依存的 NUP210 の発現を阻害し、がん細胞の増殖を抑制する2021

    • Author(s)
      近藤 寛弥、三代憲司、羽澤勝治、岩嶋友紀、小林亜紀子、小川 数馬、Richard Wong
    • Organizer
      日本生化学会北陸支部第 39 回大会
    • Related Report
      2021 Research-status Report
  • [Presentation] SARS-CoV-2 構成タンパク質 NSP9 の宿主細胞内における動態・機能に関する研究2021

    • Author(s)
      牧山桂、羽澤勝治、Richard Wong
    • Organizer
      日本生化学会北陸支部第 39 回大会
    • Related Report
      2021 Research-status Report
  • [Presentation] SARS-CoV-2由来ORF6は核膜孔複合体 RAE1とNUP98の局在変化を引き起こす2021

    • Author(s)
      加藤弘毅、小林亜紀子、羽澤勝治、Richard Wong
    • Organizer
      日本生化学会北陸支部第 39 回大会
    • Related Report
      2021 Research-status Report
  • [Presentation] 核膜孔複合体による Gene-gating 形成機序2021

    • Author(s)
      岩嶋友紀、羽澤勝治、源利成、Richard Wong
    • Organizer
      日本生化学会北陸支部第 39 回大会
    • Related Report
      2021 Research-status Report
  • [Presentation] 高速 AFM を用いた DNA インチワーミングによるヒストン H2A ラッピング動態の実時空間イメージング2021

    • Author(s)
      西出梧朗、Keesiang Lim、Mahmoud Shaaban Mohamed、小林亜紀子、羽澤勝治、中山隆宏、古寺哲幸、安藤敏夫、Richard Wong
    • Organizer
      日本生化学会北陸支部第 39 回大会
    • Related Report
      2021 Research-status Report
  • [Remarks] http://fsowonglab.w3.kanazawa-u.ac.jp/sample3.html

    • Related Report
      2022 Annual Research Report
  • [Remarks] 世界初!高速AFMで新型コロナウイルス  スパイクタンパク質の分子ナノ動態を可視化!

    • URL

      https://nanolsi.kanazawa-u.ac.jp/post-19834/

    • Related Report
      2021 Research-status Report
  • [Remarks] DNAがシャクトリムシダンス?遺伝子が核に収納される第一歩の可視化に世界で初めて成功!

    • URL

      https://nanolsi.kanazawa-u.ac.jp/post-16592/

    • Related Report
      2021 Research-status Report
  • [Remarks] 生体高分子の相分離環境を評価できる蛍光プローブの開発に成功!

    • URL

      https://www.kanazawa-u.ac.jp/rd/95286

    • Related Report
      2021 Research-status Report
  • [Remarks] Richard Wong Laboratory

    • URL

      http://fsowonglab.w3.kanazawa-u.ac.jp/

    • Related Report
      2021 Research-status Report
  • [Patent(Industrial Property Rights)] 蛍光プローブ、液相の極性及び粘性を評価する方法、並びに化合物2021

    • Inventor(s)
      羽澤 勝治, ウォング リチャード ら
    • Industrial Property Rights Holder
      羽澤 勝治, ウォング リチャード ら
    • Industrial Property Rights Type
      特許
    • Filing Date
      2021
    • Related Report
      2021 Research-status Report
  • [Patent(Industrial Property Rights)] 高速原子間力顕微鏡による細胞小器官の観察のための試料調製方法2021

    • Inventor(s)
      ウォング リチャード ら
    • Industrial Property Rights Holder
      ウォング リチャード ら
    • Industrial Property Rights Type
      特許
    • Filing Date
      2021
    • Acquisition Date
      2021
    • Related Report
      2021 Research-status Report

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Published: 2021-07-13   Modified: 2024-01-30  

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