Development of a novel technology to introduce signal transduction molecules into zebrafish embryo using membrane-translocating motif

• Project/Area Number: 24580308
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Fisheries chemistry
• Research Institution: Nihon University
• Principal Investigator: HANAZAWA Shigemasa 日本大学, 生物資源科学部, 教授 (60060258)
• Co-Investigator(Kenkyū-buntansha): MASUHIRO Yoshikazu 日本大学, 生物資源科学部, 准教授 (00336083) ; YABU Takeshi 日本大学, 生物資源科学部, 研究員 (00551756) ; IMAMURA Shintarou 独立行政法人水産総合研究センター, 研究員 (80510007)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: ゼブラフィッシュ / 細胞膜透過性ペプチド / シグナル分子 / キメラ蛋白質 / 形態形成 / 細胞分化 / 可視化 / 細胞膜透過性シグナル分子 / p53 / 転写能 / 機能発現 / 細胞膜透過性分子 / TIPE2 / 細胞膜透過性蛋白質 / 蛋白質発現

Outline of Final Research Achievements

Membrane translocating motif (MTM) composes a signal peptide sequence of hydrophobic amino acid residues, AAVLLPVLLAAP. To develop a technology to introduce directly signal transducing molecules into zebrafish embryos, we generated a recombinant MTM-tagged p53 (MTM-p53) in E.coli and then examined the transcriptional activity of the purified MTM-p53 in vitro and in vivo.
we developed in here a novel transducing technology that is able to directly introduce to zebrafish and then to functionally act MTM-p53 protein . This provides a useful approach to understand the spatiotemporal regulatory mechanism(s) of signal transducing molecules in zebrafish.

Research Products

• [Journal Article] Stress-induced ceramide generation and apoptosis via the phosphorylation and activation of nSMase1 by JNK signaling. (2015)
  • Author(s): Yabu T, Shiba H, Shibasaki Y, Nakanishi T, Imamura S, Touhata K, Yamashita M.
  • Journal Title: Cell Death Differ. Volume: 22 Issue: 2 Pages: 258-273
  • DOI: 10.1038/cdd.2014.128
  • Peer Reviewed
• [Journal Article] Peculiar monomeric interferon gammas, IFNγrel 1 and IFNγrel 2, in ginb una crucian carp. (2014)
  • Author(s): Shibasaki Y., Yabu T., Araki K., Manol N., Shiba H., Moritomo T, Nakanishi T.
  • Journal Title: FEBS Journal Volume: 281 Issue: 4 Pages: 1046-1056
  • DOI: 10.1111/febs.12666
  • Peer Reviewed / Open Access
• [Journal Article] Identification of a novel fish granzyme involved in cell-mediated immunity. (2014)
  • Author(s): Matsuura Y, Yabu T, Shiba H, Moritomo T, Nakanishi T.
  • Journal Title: Dev Comp Immunol. Volume: 150 Pages: 499-507
  • Peer Reviewed
• [Journal Article] Peculiar monomeric interferon gammas, IFNγrel 1 and IFNγrel 2, in ginbuna crucian carp. (2014)
  • Author(s): Y. Shibasaki, T. Yabu, K. Araki, N. Mano, H, Shiba, T. Moritomo, T. Nakanishi.
  • Journal Title: FEBS J. Volume: 281 Pages: 1046-1056
  • Peer Reviewed
• [Journal Article] Origin identification of dried seaweed product "nori" by PCR-RFLP analysis of Pyropia yezoensis in the internal transcribed spacer ITS-1 region. (2013)
  • Author(s): K. Touhata, A. Namikoshi, T. Suzuki, J. Iguchi, N. Mizusawa, T. Hara, S. Imamura, T. Yabu, Y. Yamashita, M. Yamashita.
  • Journal Title: Fish. Sci. Volume: 79 Pages: 865-875
  • NAID: 10031191440
• [Journal Article] Novel Selenium-Containing Compound, Mediates Detoxification Mechanisms against Methylmercury Accumulation and Toxicity in Zebrafish Embryo. (2013)
  • Author(s): M. Yamashita, Y. Yamashita, T. Suzuki, Y. Kani, N. Mizusawa, S. Imamura, K. Takemoto, T. Hara, Md. A. Hossain, T. Yabu, K. Touhata.
  • Journal Title: Mar. Biotechnol., Volume: 15 Pages: 559-570
• [Journal Article] Molecular cloning of two gonadotropin receptors in mummichog Fundulus heteroclitus and their gene expression during follicular development and maturation. (2013)
  • Author(s): M. Ohkubo, T. Yabu, M. Yamashita, A. Shimizu.
  • Journal Title: Gen. Comp. Endocrinol. Volume: 184 Pages: 75-86
• [Presentation] 膜貫通性リコンビナントGemininタンパク質は、造血幹細胞の自己複製と細胞分化を制御する新たなストラテジーを提供するだろう (2014)
  • Author(s): 黒木利知、大野芳典、安永晋一郎、竹立恭子、山藤幹茂子、舛廣善和、花澤重正、大坪素秋、瀧原義宏
  • Organizer: 第３７回日本分子生物学会
  • Place of Presentation: 神奈川県横浜市パシフィコ横浜
  • Year and Date: 2014-11-24 – 2014-11-27
• [Presentation] TIPE2はTAK1を介するH.pylori CagAシグナルを負に制御する (2014)
  • Author(s): 中野利沙、櫻井渉、大穗満隆、松本翔太、宮本梓、舛廣善和、花澤重正
  • Organizer: 第３７回日本分子生物学会
  • Place of Presentation: 神奈川県横浜市パシフィコ横浜
  • Year and Date: 2014-11-24 – 2014-11-27
• [Presentation] ストレスによるSAPK/JNKの活性化とストレス誘導性アポトーシスにおける中性スフィ ンゴミエリナーゼ１のリン酸化 (2013)
  • Author(s): 藪健史，司馬肇，柴崎康宏，中西照幸，今村伸太郎，山下倫明
  • Organizer: 日本生化学会
  • Place of Presentation: 横浜市
• [Presentation] ゼブラフィッシュ胚における細胞膜透過性シグナル分子導入法 (2012)
  • Author(s): 増見 健太郎、今村 伸太朗、薮 健史、舛廣 善和、 山下 倫明、花澤 重正
  • Organizer: 第３５回日本分子生物学会
  • Place of Presentation: 福岡市