Devolopment of a novel method to identify E3 ligase substrates

• Project/Area Number: 15K15058
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: General medical chemistry
• Research Institution: Hokkaido University
• Principal Investigator: WATANABE MASASHI 北海道大学, 医学研究科, 助教 (10632424)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: ユビキチン / ユビキチンリガーゼ

Outline of Final Research Achievements

Since ubiquitin ligase (E3) has the substrate specificity of ubiquitination, identifications of specific substrates of each E3 enzymes and of their ubiquitination sites are important for understanding various biological phenomena. In recent years, Toczyski and colleagues have proposed a ligase trap method using a fusion probe of an ubiquitin-binding domain (UBA) and an E3, and Yoshida and colleagues have proposed a TR-TUBE method combining Tandem Ubiquitin Binding Entity (TUBE) and K-εGG-specific antibody. We attempted to develop a more advanced substrate identification method by adding some improvements in their methods and succeeded in improving the sensitivity. By using this method, it is expected that the functional analysis of E3 would be accelerated.

Research Products

• [Journal Article] TRIM proteins and diseases. (2017)
  • Author(s): Watanabe, M. and Hatakeyama, S
  • Journal Title: J. Biochem. Volume: 161 Pages: 135-144
  • DOI: 10.1093/jb/mvw087
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The novel heart-specific RING finger protein 207 is involved in energy metabolism in cardiomyocytes. (2016)
  • Author(s): Mizushima, W., Takahashi , H., Watanabe, M., Kinugawa, S., Matsushima, S., Takada, S., Yokota, T., Furihata, T., Matsumoto, J., Tsuda, M., Chiba, I., Nagashima, S., Yanagi, S., Matsumoto, M., Nakayama, K.I., Tsutsui, H. and Hatakeyama, S.
  • Journal Title: J. Mol. Cell. Cardiol. Volume: 100 Pages: 43-53
  • DOI: 10.1016/j.yjmcc.2016.09.013
  • NAID: 120006360069
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] p53 represses the transcription of snRNA genes by preventing the formation of little elongation complex (2016)
  • Author(s): Anwar, D., Takahashia, H., Watanabe, M., Suzuki, M., Fukuda, S., Hatakeyama, S.
  • Journal Title: Biochim. Biophys. Acta-Gene Regul. Mech. Volume: 1859 Issue: 8 Pages: 975-982
  • DOI: 10.1016/j.bbagrm.2016.06.001
  • NAID: 120006328999
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] TRIM39 negatively regulates the NFκB-mediated signaling pathway through stabilization of Cactin (2016)
  • Author(s): Suzuki M, Watanabe M, Nakamaru Y, Takagi D, Takahashi H, Fukuda S, Hatakeyama S.
  • Journal Title: Cell Mol Life Sci. Volume: ７３ Issue: 5 Pages: 1085-1101
  • DOI: 10.1007/s00018-015-2040-x
  • NAID: 120005981342
  • Peer Reviewed / Open Access
• [Journal Article] The E3 ubiquitin ligase TRIM23 regulates adipocyte differentiation via stabilization of the adipocyte activator PPAR gamma. (2015)
  • Author(s): Watanabe, M., Takahashi, H., Saeki, Y., Ozaki, T., Itoh, S., Suzuki, M., Mizushima, W., Tanaka, K. and Hatakeyama, S.
  • Journal Title: eLIFE Volume: 4
  • DOI: 10.7554/elife.05615
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] The E3 ubiquitin ligase TRIM23 regulates adipocyte differentiation via stabilization of the adipogenic activator PPARγ (2016)
  • Author(s): 渡部 昌, 高橋秀尚, 畠山鎮次
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-12-01
  • Invited
• [Presentation] ユビキチンリガーゼTRIM23は非定型ポリユビキチン化によるPPARγの安定化を介して脂肪細胞分化を制御する (2016)
  • Author(s): 渡部 昌, 高橋秀尚, 畠山鎮次
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 東北大学川内北キャンパス （宮城県・仙台市）
  • Year and Date: 2016-09-27
• [Presentation] ユビキチンE3リガーゼTRIM23は非定型ポリユビキチン化によるPPARγの安定化を介して脂肪細胞分化を制御する (2016)
  • Author(s): 渡部 昌, 高橋秀尚, 畠山鎮次
  • Organizer: 第68回日本細胞生物学会大会
  • Place of Presentation: 京都テルサ（京都府・京都市）
  • Year and Date: 2016-06-15
• [Presentation] ユビキチンリガーゼTRIM23はPPARγの安定化を介して脂肪細胞分化を制御する (2016)
  • Author(s): 渡部 昌, 高橋秀尚, 畠山鎮次
  • Organizer: 第89回日本内分泌学会学術総会
  • Place of Presentation: 国立京都国際会館（京都府・京都市）
  • Year and Date: 2016-04-21
• [Presentation] ユビキチンリガーゼTRIM23はPPARγの安定化を介して脂肪細胞分化を制御する (2016)
  • Author(s): 渡部 昌, 高橋秀尚, 畠山鎮次
  • Organizer: 第93回日本生理学会大会
  • 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: 2016-03-24
• [Presentation] The ubiquitin E3 ligase TRIM23 regulates adipocyte differentiation via stabilization of the adipogenic activator PPARγ. (2015)
  • Author(s): Masashi Watanabe
  • Organizer: Cold Spring Harbor Laboratory Meeting on the Ubiquitin Family
  • Place of Presentation: New York(USA)
  • Year and Date: 2015-04-21
  • Int'l Joint Research
• [Book] Advances in Medicine and Biology. Volume 120 (2017)
  • Author(s): Masashi Watanabe and Shigetsugu Hatakeyama
  • Total Pages: 23
  • Publisher: Nova Science Publishers