Cross-scale measurement of nuclear pore transport system

• Project Area: New cross-scale biology
• Project/Area Number: 22H05537
• Research Category: Grant-in-Aid for Transformative Research Areas (A)
• Allocation Type: Single-year Grants
• Review Section: Transformative Research Areas, Section (III)
• Research Institution: Kanazawa University
• Principal Investigator: WONG W・R 金沢大学, ナノ生命科学研究所, 教授 (30464035)
• Project Period (FY): 2022-06-16 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000); Fiscal Year 2023: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2022: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: 核膜孔 / クロススケール / 高速AFM / 超解像顕微鏡 / 液―液相分離 / ORF6 / 脳腫瘍 / 核膜孔複合体

Outline of Research at the Start

核-細胞質を結ぶ唯一の「メゾ複雑体」輸送孔である核膜孔は、その内部に相分離から成る液滴を形成し、細胞増殖やウイルス感染応答にともなうすべてのDNA情報ネットワークを選択的に制御する。病態で変化する分子輸送の仕組みを理解するには、核膜孔内部の液滴の状態と機能相関のクロススケール核膜孔内分子構造動態の解明が必要である。本研究では、申請者らが確立してきた独自のイメージング技術により、選択的核膜分子輸送システムの基盤となる相分離のメゾ動態と機能化メカニズムを解明する。

Outline of Annual Research Achievements

核膜孔複合体（NPC: Nuclear Pore Complex）は、核内のDNAへの唯一の分子輸送経路を提供するタンパク質複合体で、核と細胞質の間の分子輸送を調節し、DNAの情報ネットワークを選択的に制御します。申請者は、高速AFM（原子間力顕微鏡）を用いて、世界で初めてFG-NUPsが形成する核膜孔内部の液滴を可視化することに成功しました。さらに、申請者らは病態制御におけるNPCの機能調節メカニズムに関する研究も進めており、細胞の環境や状態に応じて核膜孔の組成が変化し、その機能が高度化することを明らかにしました。具体的には、がん病態に特異的なFG-NUPsの発現量や翻訳後修飾が、核膜孔を介した核と細胞質間の輸送選択性を変化させ、がん病態の進行を助長する遺伝子発現パターンを確立することが示されています。しかし、液滴は動的な高次構造体であり、その安定性は環境に大きく依存します。試料の調製が難しいことに加え、相分離を調べるための技術基盤が整備されていないため、生命現象の中核を成す選択的核膜分子輸送における液滴の動態や機能メカニズム、さらには病態に伴う液滴の変化については未解明の部分が多く残っています。本研究は、核膜孔の作動原理の基盤となる相分離の理解を深めるため、核膜孔内部に形成される液滴のナノ構造動態とその機能との相関を解明することを目的としています。2022年に我々の研究成果については、Cells, BBRC, JEV誌に発表した。また、西田先生のIn-cell NMR共同研究成果については、2023年Journal of Physical Chemistry Letters誌に報告されました。

Research Progress Status

令和5年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和5年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Crafty mimicry grants nuclear pore entry to HIV (2024)
  • Author(s): Lim Keesiang、Hazawa Masaharu、Wong Richard W.
  • Journal Title: Cell Host & Microbe Volume: 32 Issue: 4 Pages: 441-442
  • DOI: 10.1016/j.chom.2024.03.004
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] An Efficient Method for Isolating and Purifying Nuclei from Mice Brain for Single-Molecule Imaging Using High-Speed Atomic Force Microscopy (2024)
  • Author(s): Qiu Yujia、Sajidah Elma Sakinatus、Kondo Sota、Narimatsu Shinnosuke、Sandira Muhammad Isman、Higashiguchi Yoshiki、Nishide Goro、Taoka Azuma、Hazawa Masaharu、Inaba Yuka、Inoue Hiroshiら
  • Journal Title: Cells Volume: 13 Issue: 3 Pages: 279-279
  • DOI: 10.3390/cells13030279
  • Peer Reviewed / Open Access
• [Journal Article] Cell-compatible isotonic freezing media enabled by thermo-responsive osmolyte-adsorption/exclusion polymer matrices (2023)
  • Author(s): Kato Yui、Matsuda Yuya、Uto Takuya、Tanaka Daisuke、Ishibashi Kojiro、Ishizaki Takeru、Ohta Akio、Kobayashi Akiko、Hazawa Masaharu、Wong Richard W.、Ninomiya Kazuaki、Takahashi Kenji、Hirata Eishu、Kuroda Kosuke
  • Journal Title: Communications Chemistry Volume: 6 Issue: 1 Pages: 260-260
  • DOI: 10.1038/s42004-023-01061-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Super-enhancer trapping by the nuclear pore via intrinsically disordered regions of proteins in squamous cell carcinoma cells (2023)
  • Author(s): Hazawa M, Ikliptikawati DK, Iwashima Y, Lin DC, Jiang Y, Qiu Y, Makiyama K, Matsumoto K, Kobayashi A, Nishide G, Keesiang L, Yoshino H, Minamoto T, Suzuki T, Kobayashi I, Meguro-Horike M, Jiang YY, Nishiuchi T, Konno H, Koeffler HP, Hosomichi K, Tajima A, Horike SI, Wong RW.
  • Journal Title: Cell Chemical Biology Volume: － Issue: 4 Pages: 792-804.e7
  • DOI: 10.1016/j.chembiol.2023.10.005
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chiral and nematic phases of flexible active filaments (2023)
  • Author(s): Dunajova Zuzana、Mateu Batirtze Prats、Radler Philipp、Lim Keesiang、Brandis Dorte、Velicky Philipp、Danzl Johann Georg、Wong Richard W.、Elgeti Jens、Hannezo Edouard、Loose Martin
  • Journal Title: Nature Physics Volume: 19 Issue: 12 Pages: 1916-1926
  • DOI: 10.1038/s41567-023-02218-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Nanoscopic Elucidation of Spontaneous Self-Assembly of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Open Reading Frame 6 (ORF6) Protein (2023)
  • Author(s): Nishide Goro、Lim Keesiang、Tamura Maiki、Kobayashi Akiko、Zhao Qingci、Hazawa Masaharu、Ando Toshio、Nishida Noritaka、Wong Richard W.
  • Journal Title: The Journal of Physical Chemistry Letters Volume: 14 Issue: 38 Pages: 8385-8396
  • DOI: 10.1021/acs.jpclett.3c01440
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Nuclear transport surveillance of p53 by nuclear pores in glioblastoma (2023)
  • Author(s): Ikliptikawati Dini Kurnia、Hirai Nozomi、Makiyama Kei、Sabit Hemragul、Kinoshita Masashi、Matsumoto Koki、Lim Keesiang、Meguro-Horike Makiko、Horike Shin-ichi、Hazawa Masaharu、Nakada Mitsutoshi、Wong Richard W.
  • Journal Title: Cell Reports Volume: 42 Issue: 8 Pages: 112882-112882
  • DOI: 10.1016/j.celrep.2023.112882
  • Peer Reviewed / Open Access / Int'l Joint Research
• [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
  • 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
  • Peer Reviewed / Open Access
• [Journal Article] Inhibition of Canonical Wnt Signaling Promotes Ex Vivo Maintenance and Proliferation of Hematopoietic Stem Cells in Zebrafish (2022)
  • 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
  • Peer Reviewed / Open Access
• [Journal Article] Discovery of a Novel Aminocyclopropenone Compound That Inhibits BRD4-Driven Nucleoporin NUP210 Expression and Attenuates Colorectal Cancer Growth (2022)
  • 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
  • Peer Reviewed / Open Access
• [Journal Article] NSP9 of SARS-CoV-2 attenuates nuclear transport by hampering nucleoporin 62 dynamics and functions in host cells (2022)
  • 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
  • Peer Reviewed / Open Access
• [Presentation] Nano imaging of genomic modality in nuclear pore territories (2023)
  • Author(s): Richard Wong
  • Organizer: The 4th “Genome Modality” Annual meeting
  • Int'l Joint Research / Invited
• [Presentation] Nano-imaging of the SARS-CoV-2 virus invasion path towards to the nuclear pore (2023)
  • Author(s): Richard Wong
  • Organizer: The 20th International Microscopy Congress
  • Int'l Joint Research / Invited
• [Presentation] 宿主の核膜孔輸送系をウィルスが乗っ取る仕組みをナノスケールで解明 (2023)
  • Author(s): Richard Wong
  • Organizer: 第96回日本生化学会大会シンポジウム
  • Invited
• [Presentation] Nano assessment of how viruses hijack host nuclear transport systems (2023)
  • Author(s): Richard Wong
  • Organizer: 学術変革領域A クロススケール新生物学 第3回領域会議
  • Invited
• [Presentation] Nano assessment of how viruses hijack the host nuclear pore (2023)
  • Author(s): Richard Wong
  • Organizer: 第23回日本蛋白質科学会年会
  • Invited
• [Presentation] Nano assessment of how viruses seize host nuclear transport systems (2022)
  • Author(s): Richard Wong
  • Organizer: 新学術創成研究機構ワークショップ
  • Invited
• [Presentation] Nano assessment of how viruses hijack host nuclear transport systems (2022)
  • Author(s): Richard Wong
  • Organizer: 15th NanoLSI Colloquium
  • Invited
• [Presentation] Nano imaging of genomic modality in nuclear pore territories (2022)
  • 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
  • Invited
• [Presentation] Nano imaging of nuclear pore dynamics in cancer cells (2022)
  • Author(s): Richard Wong
  • Organizer: 第95回日本生化学会大会シンポジウム
  • 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
  • Invited
• [Remarks] http://fsowonglab.w3.kanazawa-u.ac.jp/sample3.html