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細胞競合現象の時空間的解析とその操作

Publicly Offered Research

Project AreaCell competition: a mechanism for survival of the fittest in the multi-cellular community
Project/Area Number 17H05611
Research Category

Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionHokkaido University

Principal Investigator

大場 雄介  北海道大学, 医学研究院, 教授 (30333503)

Project Period (FY) 2017-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000)
Fiscal Year 2018: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
Fiscal Year 2017: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
Keywords蛍光バイオイメージング / 光操作 / 細胞競合
Outline of Annual Research Achievements

本研究は、光による細胞競合の誘導系を開発し、細胞競合の発生を時空間的に厳密に制御すること、同時に分子の活性化、分子間相互作用、イオン濃度変化等を蛍光バイオイメージングで観察し、winner、looser間での細胞競合が生じるための時空間的な分子動態を詳細に明らかにすることを目的に行っている。
昨年度までに、488 nmの光で分子間相互作用が誘導されるCry2-CIBの系を用いて、二つのRas活性化法(RasV12とSosを用いるもの)を構築した。この二つの系を用いて、RasV12の系のみで細胞競合が観察され、Sosによる内在性Rasの活性化では細胞競合が誘導されないことを見出した。この結果は、細胞競合において何がwinnerとlooserを決めるかという根本的課題に迫る緒になると期待された。
しかし、この系では青色光の刺激が必要なため、シグナル伝達研究に有効なフェルスター共鳴エネルギー移動(Foerster resonance energy transfer, FRET)や蛍光タンパク質再構成法(bimolecular fluorescence complementation, BiFC)等の青色光を励起光として用いるイメージング手法との併用が難しい。そこで、それらイメージング手法との併用を目指し、遠赤色光受容タンパク質phytochromeB(PhyB)とphytochrome interaction factor 3(PIF3)の結合を遠赤色光により誘導できるオプトジェネティクスの手法を取り入れた。これにより、遠赤色光照射によりSrcを細胞質から細胞膜あるいはエンドソームに移行させることに成功した。また、様々なオルガネラに局在するPhyBとPIF3を作製し、異種オルガネラ間相互作用を誘導する系も併せて開発した。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Report

(2 results)
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (25 results)

All 2019 2018 2017

All Journal Article (10 results) (of which Int'l Joint Research: 5 results,  Peer Reviewed: 10 results,  Open Access: 8 results) Presentation (14 results) Book (1 results)

  • [Journal Article] A Peptide Derived from Phosphoinositide 3-kinase Inhibits Endocytosis and Influenza Virus Infection2019

    • Author(s)
      Fujioka Y, Satoh AO, Horiuchi K, Fujioka M, Tsutsumi K, Sasaki J, Nepal P, Kashiwagi S, Paudel S, Nishide S, Nanbo A, Sasaki T, Ohba Y
    • Journal Title

      Cell Structure and Function

      Volume: 44 Issue: 1 Pages: 61-74

    • DOI

      10.1247/csf.19001

    • NAID

      130007636814

    • ISSN
      0386-7196, 1347-3700
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] A Sialylated Voltage-Dependent Ca2+ Channel Binds Hemagglutinin and Mediates Influenza A Virus Entry into Mammalian Cells2018

    • Author(s)
      Fujioka Y, Nishide S, Ose T, Suzuki T, Kato I, Fukuhara H, Fujioka M, Horiuchi K, Satoh AO, Nepal P, Kashiwagi S, Wang J, Horiguchi M, Sato Y, Paudel S, Nanbo A, Miyazaki T, Hasegawa H, Maenaka K, Ohba Y
    • Journal Title

      Cell Host Microbe

      Volume: 23 Issue: 6 Pages: 809-818

    • DOI

      10.1016/j.chom.2018.04.015

    • NAID

      120006643588

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] The Role of Transforming Growth Factor β in Cell-to-Cell Contact-Mediated Epstein-Barr Virus Transmission2018

    • Author(s)
      Nanbo A, Ohashi M, Yoshiyama H, Ohba Y
    • Journal Title

      Front Microbiol

      Volume: 9 Pages: 984-984

    • DOI

      10.3389/fmicb.2018.00984

    • NAID

      120006489803

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Infection of Epstein?Barr Virus in Type III Latency Modulates Biogenesis of Exosomes and the Expression Profile of Exosomal miRNAs in the Burkitt Lymphoma Mutu Cell Lines2018

    • Author(s)
      Nanbo Asuka、Katano Harutaka、Kataoka Michiyo、Hoshina Shiho、Sekizuka Tsuyoshi、Kuroda Makoto、Ohba Yusuke
    • Journal Title

      Cancers

      Volume: 10 Issue: 7 Pages: 237-237

    • DOI

      10.3390/cancers10070237

    • NAID

      120006491456

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Budding of Ebola Virus Particles Requires the Rab11-Dependent Endocytic Recycling Pathway2018

    • Author(s)
      Nanbo A, Ohba Y
    • Journal Title

      J Infect Dis

      Volume: 218 Issue: suppl_5 Pages: S388-S396

    • DOI

      10.1093/infdis/jiy460

    • NAID

      120006773327

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Epstein-Barr Virus Acquires Its Final Envelope on Intracellular Compartments With Golgi Markers2018

    • Author(s)
      Nanbo Asuka、Noda Takeshi、Ohba Yusuke
    • Journal Title

      Front Microbiol

      Volume: 9 Pages: 454-454

    • DOI

      10.3389/fmicb.2018.00454

    • NAID

      120006416029

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Dermoscopic evaluation for skin grafts after surgery; neo-vascularization correlates with survival of skin grafts: A prospective study2018

    • Author(s)
      Kitamura Shinya、Yanagi Teruki、Inamura-Takashima Yuka、Imafuku Keisuke、Hata Hiroo、Fujioka Yoichiro、Ohba Yusuke、Shimizu Hiroshi
    • Journal Title

      J Dermatol Sci

      Volume: 印刷中 Issue: 2 Pages: 213-216

    • DOI

      10.1016/j.jdermsci.2018.01.014

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Ebola virus requires a host scramblase for externalization of phosphatidylserine on the surface of viral particles2018

    • Author(s)
      Nanbo Asuka、Maruyama Junki、Imai Masaki、Ujie Michiko、Fujioka Yoichiro、Nishide Shinya、Takada Ayato、Ohba Yusuke、Kawaoka Yoshihiro
    • Journal Title

      PLoS Pathog

      Volume: 14 Issue: 1 Pages: e1006848-e1006848

    • DOI

      10.1371/journal.ppat.1006848

    • NAID

      120006416019

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Cell competition with normal epithelial cells promotes apical extrusion of transformed cells through metabolic changes.2017

    • Author(s)
      Shunsuke Kon, et al.
    • Journal Title

      Nature Cell Biology

      Volume: 印刷中 Issue: 5 Pages: 530-541

    • DOI

      10.1038/ncb3509

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Rab5-regulated endocytosis plays a crucial role in apical extrusion of transformed cells.2017

    • Author(s)
      S. Saitoh, T. Maruyama, Y. Yako, M. Kajita, Y. Fujioka, Y. Ohba, N. Kasai, N. Sugama, S. Kon, S. Ishikawa, T. Hayashi, T. Yamazaki, M. Tada and Y. Fujita.
    • Journal Title

      Proc. Natl. Acad. Sci. U.S.A.

      Volume: 114 Issue: 12

    • DOI

      10.1073/pnas.1602349114

    • NAID

      120006346108

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Fluorescence Imaging of membrane dynamics and intracellular signaling2019

    • Author(s)
      Yusuke Ohba
    • Organizer
      The 9th Federation of Asian and Oceanian Physiological Societies Congress (FAOPS2019) in conjunction with the 96th Annual Meeting of the Physiological Society of Japan
    • Related Report
      2018 Annual Research Report
  • [Presentation] A PI3K-derived peptide inhibits clathrin-independent endocytosis and influenza virus infection2018

    • Author(s)
      Yoichiro Fujioka, Aya O Satoh, Kosui Horiuchi, Mari Fujioka, Sarad Paudel, Asuka Nanbo, Yusuke Ohba
    • Organizer
      第70回日本細胞生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Involvement of actin dynamics in the endocytic process revealed by fast-scanning atomic force microscopy2018

    • Author(s)
      Aiko Yoshida, Nobuaki Sakai, Yoshitsugu Uekusa, Shige H Yoshimura, Yusuke Ohba
    • Organizer
      第70回日本細胞生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] The role of an Na,K-ATPase in spatiotemporal regulation of Ras- PI3K signaling and endocytosis2018

    • Author(s)
      Sayaka Kashiwagi, Yoichiro Fujioka, Kosui Horiuchi, Aya O Satoh, Prabha Nepal, Aiko Yoshida, Sarad Paudel, Asuka Nanbo, Yusuke Ohba
    • Organizer
      第70回日本細胞生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] A mitochondrial outer membrane protein is involved in the regulation of Ras-PI3K signaling-mediated endocytosis2018

    • Author(s)
      Aya O Satoh, Yoichiro Fujioka, Kosui Horiuchi, Prabha Nepal, Sayaka Kashiwagi, Aiko Yoshida, Mari Fujioka, Sarad Paudel, Asuka Nanbo, Yusuke Ohba
    • Organizer
      第70回日本細胞生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Development of FRET-based biosensors for measuring tyrosine kinase activity in living cells2018

    • Author(s)
      Mari Fujioka, Yoichiro Fujioka, Aya O Satoh, Prabha Nepal, Sayaka Kashiwagi, Aiko Yoshida, Sarad Paudel, Asuka Nanbo, Yusuke Ohba
    • Organizer
      第70回日本細胞生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] The Role of tubulin in the regulation of endocytosis mediated by Ras-PI3K signaling2018

    • Author(s)
      Sarad Paudel, Yoichiro Fujioka, Aya O. Satoh, Mari Fujioka, Kosui Horiuchi, Prabha Nepal, Sayaka Kashiwagi, Aiko Yoshida, Asuka Nanbo, Yusuke Ohba
    • Organizer
      第70回日本細胞生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Role of inner mitochondrial membrane proteins in the regulation of endocytosis mediated by Ras-PI3K signaling2018

    • Author(s)
      Prabha Nepal, Yoichiro Fujioka, Aya O. Satoh, Kosui Horiuchi, Sarad Paudel, Sayaka Kashiwagi, Aiko Yoshida, Mari Fujioka, Asuka Nanbo, Yusuke Ohba
    • Organizer
      第70回日本細胞生物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 細胞内シグナル伝達の可視化と光による操作2017

    • Author(s)
      大場雄介
    • Organizer
      第40回日本分子生物学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] PI3K 由来ペプチドによるエンドサイトーシスとインフルエンザウイルス感染の抑制2017

    • Author(s)
      藤岡容一朗、佐藤絢、堀内浩水、藤岡真理、パウデル サラド、西出真也、南保明日香、大場雄介
    • Organizer
      第40回日本分子生物学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 高速原子間力顕微鏡を用いた細胞表面膜構造動態のライブセルイメージング2017

    • Author(s)
      吉田藍子、酒井信明、植草良嗣、粂田昌宏、吉村成弘、大場雄介
    • Organizer
      第40回日本分子生物学会年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] インフルエンザウイルス細胞侵入において鍵となる宿主タンパク質の同定2017

    • Author(s)
      藤岡容一朗、西出真也、尾瀬農之、加藤いづみ、福原秀雄、藤岡真理、堀内浩水、佐藤絢、Prabha Nepal、柏木彩花、Jing Wang、堀口美香、Sarad Paudel、南保明日香、宮崎忠昭、前仲勝実、大場雄介
    • Organizer
      第69回日本細胞生物学会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Ras-PI3K複合体によるエンドサイトーシスの制御因子の探索と機能解析2017

    • Author(s)
      堀内浩水、藤岡容一朗、佐藤絢、Prabha Nepal、Jing Wang、堀口美香、Sarad Paudel、西出真也、南保明日香、小布施力史、大場雄介
    • Organizer
      第69回日本細胞生物学会
    • Related Report
      2017 Annual Research Report
  • [Presentation] PI3K 由来ペプチドによるインフルエンザウイルス感染の抑制2017

    • Author(s)
      佐藤絢、藤岡容一朗、堀内浩水、藤岡真理、パウデル サラド、西出真也、南保明日香、大場雄介
    • Organizer
      第69回日本細胞生物学会
    • Related Report
      2017 Annual Research Report
  • [Book] Methods in Molecular Biology, SH2 domain2017

    • Author(s)
      Mari Fujioka、Yumi Asano、Shigeyuki Nakada、Yusuke Ohba
    • Total Pages
      555
    • Publisher
      Humana Press, New York, NY
    • ISBN
      9781493967629
    • Related Report
      2017 Annual Research Report

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Published: 2017-04-28   Modified: 2019-12-27  

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