Intracellular signaling machinery that regulates endocytosis and uptake of exogenous factors(Fostering Joint International Research)

• Project/Area Number: 15KK0290
• Research Category: Fund for the Promotion of Joint International Research (Fostering Joint International Research)
• Allocation Type: Multi-year Fund
• Research Field: General physiology
• Research Institution: Hokkaido University
• Principal Investigator: Ohba Yusuke 北海道大学, 医学研究院, 教授 (30333503)
• Research Collaborator: Tani Tomomi Marine Biological Laboratory, Eugene Bell Center for Regenerative Biology and Tissue Engineering, Research Associate
• Project Period (FY): 2016 – 2018
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥14,300,000 (Direct Cost: ¥11,000,000、Indirect Cost: ¥3,300,000)
• Keywords: 蛍光イメージング / シグナル伝達 / エンドサイトーシス

Outline of Final Research Achievements

We have developed a technology to obtain information on "angle" in biological phenomena by measuring the "orientation" of molecules using the properties of polarization and anisotropy of fluorescence. In particular, we established a method to detect the changes in the angle (curvature) of cell membranes by translating it to changes in fluorescence intensity corresponding to alteration of the molecular orientation. With these advantages, the curvature of the plasma membrane and its change, which had been visible only observable with electron microscopy, can be optically observed at a high time resolution (second level), making possible to visualize cell membrane deformation in the very early stage of endocytosis.

Academic Significance and Societal Importance of the Research Achievements

本研究成果は、エンドサイトーシスの超初期過程の細胞膜変形の分子メカニズムを解明に資する技術革新といえる。細胞膜の変形は100 nm程度以下の領域で生じる現象で、現存する超解像顕微鏡でも時間分解能を伴うダイナミクスの観察は不可能であり、「向き」に着目することで従来技術の限界を超える観察が可能となった。今後は、本研究成果を基盤に、「角度」と「向き」に着目した新しい生物学の新領域の開拓を目指したい。

Research Products

• [Int'l Joint Research] Marine Biological Laboratory(米国) (2018)
• [Journal Article] A Peptide Derived from Phosphoinositide 3-kinase Inhibits Endocytosis and Influenza Virus Infection (2019)
  • 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
  • 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 Cells (2018)
  • 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
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The Role of Transforming Growth Factor β in Cell-to-Cell Contact-Mediated Epstein-Barr Virus Transmission (2018)
  • 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
  • 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 Lines (2018)
  • 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
  • Peer Reviewed / Open Access
• [Journal Article] Budding of Ebola Virus Particles Requires the Rab11-Dependent Endocytic Recycling Pathway (2018)
  • 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
  • Peer Reviewed / Open Access
• [Journal Article] Epstein-Barr Virus Acquires Its Final Envelope on Intracellular Compartments With Golgi Markers (2018)
  • 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
  • Peer Reviewed / Open Access
• [Journal Article] Dermoscopic evaluation for skin grafts after surgery; neo-vascularization correlates with survival of skin grafts: A prospective study (2018)
  • 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
  • Peer Reviewed
• [Journal Article] Ebola virus requires a host scramblase for externalization of phosphatidylserine on the surface of viral particles (2018)
  • 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
  • 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
  • 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
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Improved FRET Biosensor for the Measurement of BCR-ABL Activity in Chronic Myeloid Leukemia Cells (2017)
  • Author(s): M. Horiguchi, M. Fujioka, T. Kondo, Y. Fujioka, X. Li, K. Horiuchi, A.O. Satoh, P. Nepal, S. Nishide, A. Nanbo, T. Teshima and Y. Ohba.
  • Journal Title: Cell Structure and Function Volume: 42 Issue: 1 Pages: 15-26
  • DOI: 10.1247/csf.16019
  • NAID: 130005308488
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed / Open Access
• [Journal Article] A leukemogenic kinase, FIP1L1-PDGFRA, and a SUMO E3 ligase, PIAS1, form a positive crosstalk via their enzymatic activities. (2017)
  • Author(s): M. Ibata, J. Iwasaki, Y. Fujioka, K. Nakagawa, S. Darmanin, M. Onozawa, D. Hashimoto, Y. Ohba, S. Hatakeyama, T. Teshima and T. Kondo.
  • Journal Title: Cancer Sci. Volume: 108 Issue: 2 Pages: 200-207
  • DOI: 10.1111/cas.13129
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Epstein-Barr virus exploits host endocytic machinery for cell-to-cell viral transmission rather than a virological synapse. (2016)
  • Author(s): A. Nanbo, K. Kachi, H. Yoshiyama and Y. Ohba.
  • Journal Title: J. Gen. Virol. Volume: 97 Issue: 11 Pages: 2989-3006
  • DOI: 10.1099/jgv.0.000605
  • NAID: 120006360051
  • Peer Reviewed / Open Access
• [Journal Article] Tumour endothelial cells in high metastatic tumours promote metastasis via epigenetic dysregulation of biglycan. (2016)
  • Author(s): N. Maishi, Y. Ohba, K. Akiyama, N. Ohga, J.I. Hamada, H. Nagao-Kitamoto, M. Alam, K. Yamamoto, T. Kawamoto, N. Inoue, A. Taketomi, M. Shindoh, Y. Hida and K. Hida.
  • Journal Title: Sci. Rep. Volume: 6 Issue: 1 Pages: 28039-28039
  • DOI: 10.1038/srep28039
  • NAID: 120005819304
  • Peer Reviewed / Open Access
• [Journal Article] Lypd8 promotes the segregation of flagellated microbiota and colonic epithelia. (2016)
  • Author(s): Okumura R, Kurakawa T, Nakano T, Kayama H, Kinoshita M, Motoka D, Gotoh K, Kimura T, Kamiyama N, Kusu T, Ueda Y, Wu H, Iijima H, Barman S, Osawwa H, Matsuno H, Nishimura J, Ohba Y, Nakamura S, Iida T, Yamamoto M, Umemoto E, Sano K, and Takeda K
  • Journal Title: Nature Volume: 532 Issue: 7597 Pages: 117-121
  • DOI: 10.1038/nature17406
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Receptor activator of NF-κB ligand induces cell adhesion and integrin α2 expression via NF-κB in head and neck cancers (2016)
  • Author(s): T.Yamada, M. Tsuda, T. Wagatsuma, Y. Fujioka, M. Fujioka, A. O. Satoh, K. Horiuchi, S. Nishide, A. Nanbo, Y. Totsuka, H. Haga, S. Tanaka, M. Shindoh, and Y. Ohba
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 23545-23545
  • DOI: 10.1038/srep23545
  • NAID: 120005753715
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Attenuation of ligand-induced activation of angiotensin II type 1 receptor signaling by the type 2 receptor via protein kinase C (2016)
  • Author(s): Inuzuka T, Fujioka Y, Tsuda M, Fujioka M, Satoh AO, Horiuchi K, Nishide S, Nanbo A, Tanaka S, Ohba Y
  • Journal Title: Sci Rep Volume: 6 Issue: 1 Pages: 21613-21613
  • DOI: 10.1038/srep21613
  • NAID: 120005722549
  • Peer Reviewed / Open Access
• [Journal Article] A role of the sphingosine-1-phosphate (S1P)- S1P receptor 2 pathway in Epithelial Defense Against Cancer (EDAC) (2015)
  • Author(s): Yamamoto, S., Yako, Y., Fujioka, Y., Kajita, M., Kameyama, T., Kon, S., Ishikawa, S., Ohba, Y., Ohno, Y., Kihara, A., and Fujita, Y.
  • Journal Title: Molecular Biology of the Cell Volume: 27（3） Issue: 3 Pages: 491-499
  • DOI: 10.1091/mbc.e15-03-0161
  • NAID: 120005740332
  • Peer Reviewed / Open Access
• [Presentation] Fluorescence Imaging of membrane dynamics and intracellular signaling (2019)
  • 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
• [Presentation] A PI3K-derived peptide inhibits clathrin-independent endocytosis and influenza virus infection (2018)
  • 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
• [Presentation] Involvement of actin dynamics in the endocytic process revealed by fast-scanning atomic force microscopy (2018)
  • 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
• [Presentation] The role of an Na,K-ATPase in spatiotemporal regulation of Ras- PI3K signaling and endocytosis (2018)
  • 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
• [Presentation] A mitochondrial outer membrane protein is involved in the regulation of Ras-PI3K signaling-mediated endocytosis (2018)
  • 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
• [Presentation] Development of FRET-based biosensors for measuring tyrosine kinase activity in living cells (2018)
  • 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
• [Presentation] The Role of tubulin in the regulation of endocytosis mediated by Ras-PI3K signaling (2018)
  • 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
• [Presentation] Role of inner mitochondrial membrane proteins in the regulation of endocytosis mediated by Ras-PI3K signaling (2018)
  • 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
• [Presentation] 蛍光イメージングを用いた細胞内シグナル伝達によるエンドサイトーシスの制御機構の解析 (2017)
  • Author(s): 大場雄介、藤岡容一朗、田端和仁、西出真也、南保明日香、野地博行
  • Organizer: 第122回解剖学会
  • Place of Presentation: 長崎大学坂本キャンパス（長崎県・長崎市）
  • Year and Date: 2017-03-28
  • Invited
• [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
  • Invited
• [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
• [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
• [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
• [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
• [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
• [Presentation] 蛍光イメージングで紐解くインフルエンザウイルス感染の分子基盤 (2016)
  • Author(s): 大場 雄介、藤岡 容一朗、西出 真也、南保 明日香
  • Organizer: 第54回日本生物物理学会年会
  • Place of Presentation: つくば国際会議場(茨城県・つくば市)
  • Year and Date: 2016-11-25
  • Invited
• [Presentation] 慢性骨髄性白血病分子標的治療薬薬効評価のためのFRETバイオセンサーの開発と改良 (2016)
  • Author(s): 大場雄介、近藤健、豊嶋崇徳
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-10-06
• [Presentation] 生きた白血病細胞でBCR-ABL活性を精密測定するめのFRETバイオセンサーの改良 (2016)
  • Author(s): 大場雄介、藤岡真理、堀口美香、近藤健、豊嶋崇徳
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台国際センター 東北大学川内北キャンパス（宮城県・仙台市）
  • Year and Date: 2016-09-25
• [Presentation] ライブセル蛍光イメージングの口腔疾患予防への応用 (2016)
  • Author(s): 西出 真也、藤岡 容一朗、堀内 浩水、堀口 美香、佐藤 絢、 Prabha Nepal、王 セイ、南保 明日香、大場 雄介
  • Organizer: 第58回歯科基礎医学会学術大会
  • Place of Presentation: 札幌コンベンションセンター(北海道・札幌市）
  • Year and Date: 2016-08-24
• [Presentation] インフルエンザウイルス粒子の取り込みを制御するシグナル伝達機構 (2016)
  • Author(s): 大場 雄介
  • Organizer: 第6回北海道探索病理学研究シンポジウム
  • Place of Presentation: ニューオータニイン札幌 （北海道・札幌市）
  • Invited
• [Book] Methods in Molecular Biology, SH2 domain (2017)
  • Author(s): Mari Fujioka、Yumi Asano、Shigeyuki Nakada、Yusuke Ohba
  • Total Pages: 555
  • Publisher: Humana Press, New York, NY
• [Book] SH2 domain-based FRET biosensor for measuring BCR-ABL activity in living CML cells. (2017)
  • Author(s): Fujioka M, Asano Y, Nakada S and Ohba Y.
  • Total Pages: 22
  • Publisher: Springer