Exploitation of a tool for analyzing molecular interactions on the plasma membrane by using membrane-bound antibodies

• Project/Area Number: 23657142
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Developmental biology
• Research Institution: The University of Tokyo
• Principal Investigator: TAIRA Masanori 東京大学, 大学院・理学系研究科, 准教授 (60150083)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2011: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
• Keywords: 膜結合型抗体 / 一本鎖抗体 / 細胞間架橋 / 力学的相互作用 / 抗原抗体反応 / 分子間相互作用 / 細胞間張力 / 張力センサーモジュール / 細胞運動

Research Abstract

We cloned cDNAs for anti-HA and anti-Myc monoclonal antibodies, and made membrane-bound antibody constructs, which were verified to function as antibodies against HA- and Myc-tagged proteins. We further made single-stranded antibodies, which were verified to function as antibodies. Using membrane-bound antibodies, we next tried to measure the tension between two adjacent cells in Xenopus embryos by using the FRET-based tension sensor module (TSMod). In addition, we found that development of Xenopus gastrulae is affected when cells are bridged with the membrane-bound antibodies and membrane-bound antigens. This bridging method can be used as a tool for investigation of intercellular plasticity during morphogenesis.

Research Products

• [Journal Article] Dynamic in vivo binding of transcription factors to cis-regulatory modules of cer and gsc in the stepwise formation of the Spemann-Mangold organizer. (2012)
  • Author(s): Sudou, N., Yamamoto, S., Ogino, H. and Taira, M.
  • Journal Title: Development Volume: 139 Pages: 1651-1661
• [Journal Article] The medaka enhancer mutant for zic1/zic4 provides molecular insights into teleost caudal fin evolution. (2012)
  • Author(s): Moriyama, Y., Kawanishi, T., Nakamura, R., Tsukahara, T., Sumiyama, K., Suster, M., Kawakami, K., Toyoda, A., Fujiyama, A., Yasuoka, Y., Nagao, Y., Sawatari, E., Shimikzu, A., Wakamatsu, Y., Hibi, M., Taira, M., Okabe, M., Naruse, K., Hashimoto, H., Shimada, A., Takeda, H.
  • Journal Title: Curr. Biol. Volume: 22 Pages: 601-607
• [Journal Article] Secreted Wnt "inhibitors" are not just inhibitors : Regulation of extracellular Wnt by secreted Frizzled-related proteins (2011)
  • Author(s): Yusuke Mii
  • Journal Title: Development, Growth & Differentiation Volume: 53 Issue: 8 Pages: 911-923
  • DOI: 10.1111/j.1440-169x.2011.01299.x
  • Peer Reviewed
• [Presentation] Heparan sulfate nanostructures regulate extracellular Wnt distribution and act asa core for Wnt/Dishevelled signalosome formation (2012)
  • Author(s): Masanori Taira and Yusuke Mii
  • Organizer: 14th International Xenopus Conference
  • Place of Presentation: Giens Peninsula (France)
• [Presentation] Heparan sulfate nanostructures regulate extracellular Wnt distribution and act as a core for Wnt/Dishevelled signalosome formation (2012)
  • Author(s): Yusuke Mii
  • Organizer: 14th International Xenopus Conference
  • Place of Presentation: Giens Peninsula (France)
• [Presentation] Secreted Frizzled-related proteins (sFRPs) regulate extracellular distribution of Wnt ligands via interactions with heparan sulfate (2012)
  • Author(s): Yusuke Mii, Kenichi Nakazato, Atsushi Mochizuki and Masanori Taira
  • Organizer: 日本発生生物学会第44回大会
  • Place of Presentation: 沖縄コンベンションセンター
• [Presentation] The molecular entity of positional information revealed by ChIP- sequence analysis for Lim1 and Otx2 in the head organizer and its application for evolutionary analysis (2012)
  • Author(s): Yuuri Yasuoka, Norihiro Sudou, Shuji Takahashi, Yukiko Tando, Yutaka Suzuki, Sumio Sugano, Kaoru, Kubokawa, Makoto Asashima and Masanori Taira
  • Organizer: 日本発生生物学会第44回大会
  • Place of Presentation: 沖縄コンベンションセンター
• [Presentation] Frzbregulates extracellular distribution of Wnt8 via interactions with heparan sulfate (2011)
  • Author(s): Yusuke Mii, Kenichi Nakazato, AtsushiMochizuki and Masanori Taira
  • Organizer: Wnt 2011 Meeting, UCLA
  • Place of Presentation: California, U.S.A.
• [Presentation] Secreted Frizzled-related proteins (sFRPs) regulate extracellular distribution of Wnt ligands via interactions with heparan sulfate.(分泌性 Frizzled 関連蛋白質 (sFRPs) はヘパラン硫酸を介して細胞外の Wnt リガンドの分布を制御する。) (2011)
  • Author(s): Yusuke Mii, Kenichi Nakazato, Atsushi Mochizuki and Masanori Taira
  • Organizer: 日本発生生物学会第44回大会
  • Place of Presentation: 沖縄コンベンションセンター
• [Presentation] Xenopusの眼の初期形成に関わるOtx2の活性調節におけるC2H2型Znフィンガー蛋白質Zbtb11とZnf668の役割 (2011)
  • Author(s): 佐藤夢子、柴野卓志、儘田博志、安岡有理、高橋秀治、浅島誠、平良眞規
  • Organizer: 第34回分子生物学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] ChIP-seq法を用いた内胚葉ChIP-seq法を用いた内胚葉と中胚葉形成におけるVegTとMix1の役割の解析ChIP-seq法を用いた内胚葉と中胚葉形成におけるVegTとMix1の役割の解析と中胚葉形成におけるVegTとMix1の役割の解析 (2011)
  • Author(s): 桐ケ谷嘉章、安岡有理、鈴木穣、高橋秀治、浅島誠、菅野純夫、平良眞規
  • Organizer: 第34回分子生物学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] Direct contributions of Otx2 as a positional tag to global gene regulation in the head organizer in Xenopus embryos (2011)
  • Author(s): 平良眞規
  • Organizer: 国際生物学賞記念国際シンポジウム（招待講演）(招待講演)
  • Place of Presentation: 京都ガーデンパレス
• [Presentation] Frzb regulates extracellular distribution of Wnt8 via interactions with heparan sulfate (2011)
  • Author(s): Yusuke Mii, Kenichi Nakazato, Atsushi Mochizuki and Masanori Taira
  • Organizer: Wnt Meeting 2011
  • Place of Presentation: UCLA, California, U.S.A.
• [Remarks]
  • URL: http://www.biol.s.u-tokyo.ac.jp/users/lmb/index.php-Home