Regulation of membrane traffic by ubiquitination

• Project/Area Number: 21570213
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Cell biology
• Research Institution: 防衛大学校
• Principal Investigator: ICHIMURA Tohru 防衛大学校, 応用科学群, 教授 (50213012)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2009: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: ユビキチン / リン酸化 / シグナル伝達 / 14-3-3タンパク質 / メンブレントラフィック / TRIM32 / TRiM32

Research Abstract

The relevance of the ubiquitin(Ub)-proteasome system is widely accepted in a variety of cell system. We previously reported a MS-based proteomic technology involving multi-step immunoaffinity purification to characterize signaling complexes in a large scale. In this study we described a novel procedure that combined the technology with newly developed Ub derivatives that inserted a His tag within its C-terminal region. Using this procedure, we successfully identified in vivo ubiquitination sites of over 200 proteins with high confidence. In another experiment, we investigated the crosstalk between phosphorylation and ubiquitination. We found that the ubiquitin E3 ligase TRIM32 binds 14-3-3 proteins in a PKA-catalyzed phosphorylation-dependent manner. Subsequent studies demonstrated that this interaction negatively regulates the catalytic activity and intracellular transport of TRIM32.The findings described herein constitute the first evidence for the regulatory mechanism of the TRIM member.

Research Products

• [Journal Article] Interaction of nucleosome assembly proteins abolishes nuclear localization of DGKζby attenuating its association with importins (2011)
  • Author(s): Okada M., Hozumi Y., Ichimura T., Tanaka T., Hasegawa H., Yamamoto M., Takahashi N., Iseki K., Yagisawa H., Shinkawa T., Isobe T., & Goto K.
  • Journal Title: Exp. Cell Res Volume: 317 Pages: 2853-2863
  • URL: http://www.ncbi.nlm.nih.gov/pubmed/21996351
  • Peer Reviewed
• [Journal Article] NAP Interaction of nucleosome assembly proteins abolishes nuclear localization of DGKζ by attenuating its association with importins (2011)
  • Author(s): Okada M, Hozumi Y, Ichimura T, Tanaka T, Hasegawa H, Yamamoto M, Takahashi N, Iseki K, Yagisawa H, Shinkawa T, Isobe T, Goto K.
  • Journal Title: Exp Cell Res Volume: 317 Issue: 20 Pages: 2853-63
  • DOI: 10.1016/j.yexcr.2011.09.014
  • Peer Reviewed
• [Journal Article] Identification of a suppressive mechanism for Hedgehogsignaling through a novel interaction of Gli with 14-3-3 (2010)
  • Author(s): Asaoka, Y., Kanai, F., Ichimura, T., Tateishi, K., Tanaka, Y., Ohta, M., Seto, M., Tada, M., Ijichi, H., Ikenoue, T., Kawabe, T., Isobe, T., Yaffe, M. B., & Omata, M.
  • Journal Title: J. Biol. Chem Volume: 285 Pages: 4185-4194
  • URL: http://www.ncbi.nlm.nih.gov/pubmed/19996099
  • Peer Reviewed
• [Journal Article] E6AP ubiquitin ligase mediates ubiquitin-dependent 1 degradation of peroxiredoxin 1 (2010)
  • Author(s): Nasu, J., Murakami, K., Miyagawa, S., Yamashita, R., Ichimura, T., Wakita, T., Hotta, H., Miyamura, T., Suzuki, T., Satoh, T. & Shoji, I.
  • Journal Title: J. Cell. Biochem. Volume: 111 Pages: 676-685
  • URL: http://www.ncbi.nlm.nih.gov/pubmed/20589759
  • Peer Reviewed
• [Journal Article] E6AP ubiquitin ligase mediates ubiquitin-dependent degradation of peroxiredoxin 1 (2010)
  • Author(s): Nasu Junichi
  • Journal Title: J.Cell Biochem. Volume: VOL.111 Pages: 678-685
  • Peer Reviewed
• [Journal Article] Two beclin-binding proteins, Atg14L and Rubicon reciprocally regulate autophagy at different stages (2009)
  • Author(s): Matsunaga, K., Saitoh, T., Tabata, K., Omori, H., Satoh, T., Kurotori, N., Maejima, I., Shirahama-Noda, K., Ichimura, T., Isobe, T., Akira, S., Noda, T., & Yoshimori, T.
  • Journal Title: Nat Cell Biol. Volume: 11 Pages: 385-396
  • URL: http://www.ncbi.nlm.nih.gov/pubmed/19270696
  • Peer Reviewed
• [Journal Article] Two beclin-binding proteins, Atg14L and Rubicon, reciprocally regulate autophagy at different stages (2009)
  • Author(s): Matsubaga Koichi
  • Journal Title: Nat Cell Biol VOL.11 Pages: 385-396
  • Peer Reviewed
• [Journal Article] Identification of a suppressive mechanism for Hedgehog signaling through a novel interaction of Gli with 14-3-3. (2009)
  • Author(s): Asaoka Yoshinari
  • Journal Title: J.Biol.Chem. VOL.285 Pages: 4185-4194
  • Peer Reviewed
• [Presentation] A Regulatory Mechanism for Tripartite-motif Protein 32 Revealed by Quantitative 14-3-3 Interactome Analysis in PKA signaling Pathways (2011)
  • Author(s): T. Ichimura, I. Shoji, and N. Hachiya
  • Organizer: 第34回日本分子生物学会
  • Place of Presentation: パシフィコ横浜:神奈川県
  • Year and Date: 2011-12-15
• [Presentation] A regulatory mechanism for tripartite-motif protein32 revealed by quantitative 14-3-3 interactome analysis in PKA signaling pathways (2011)
  • Author(s): Tohru Ichimura
  • Organizer: 第34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)
  • Year and Date: 2011-12-03
• [Presentation] PKA/14-3-3によるユビキチンリガーゼTRIM32の制御 (2011)
  • Author(s): 市村徹,八谷如美,小俣結子,畠山鎮次勝二郁夫
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都国際会館:京都府
  • Year and Date: 2011-09-24
• [Presentation] PKA/14-3-3によるユビキチンリガーゼTRIM32の制御 (2011)
  • Author(s): 市村徹
  • Organizer: 第84回日本生化学会大会合同大会
  • Place of Presentation: 国立京都国際会館(京都府)
  • Year and Date: 2011-09-24
• [Presentation] E6AP ubiquitin ligase mediatesubiquitin-dependent degradation of peroxiredoxin 1 (2010)
  • Author(s): S. Ikuo, J. Nasu, K. Murakami, S. Miyagawa, R. Yamashita, T. Ichimura, T. Wakita, Y. Ide, H. Hotta, T. Miyamura, T. Suzuki, and T. Satoh
  • Organizer: 第33回日本分子生物学会
  • Place of Presentation: 神戸ポートアイランド:兵庫県
  • Year and Date: 2010-12-09
• [Presentation] E6AP ubiquitin ligase mediates ubiquitin-dependent degradation of peroxiredoxin 1 (2010)
  • Author(s): 勝二郁夫
  • Organizer: 第32回日本分子生物学会第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド(兵庫県)
  • Year and Date: 2010-12-09
• [Presentation] TRIM-NHLタンパク質のリン酸化による調節 (2009)
  • Author(s): 吉田順,市村徹,垣内一恵
  • Organizer: 第32回日本分子生物学会
  • Place of Presentation: パシフィコ横浜:神奈川県
  • Year and Date: 2009-12-11
• [Presentation] TRIM-NHLタンパク質TRIM32のリン酸化による調節 (2009)
  • Author(s): 吉田順
  • Organizer: 第32回日本分子生物学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2009-12-09
• [Presentation] 二つの新規Beclin1結合タンパク質であるAtg14LとRubiconは、それぞれオートファジーを正負に制御する (2009)
  • Author(s): 松永耕一,斎藤達哉,田端桂介,大森弘子,佐藤荘,黒鳥直樹,前島郁子,野田桂苗,市村徹,礒辺俊明,審良静男,野田健司,吉森保
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸ポートアイランド:兵庫県
  • Year and Date: 2009-10-24
• [Presentation] リン酸化とユビキチン化のクロストーク (2009)
  • Author(s): 吉田順、市村徹
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸ポートアイランド:兵庫県
  • Year and Date: 2009-10-23
• [Presentation] E3ユビキチンリガーゼTrim32と14-3-3タンパク質の相互作用と機能解析 (2009)
  • Author(s): 吉田順
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2009-10-21