Regulation of ubiquitin homeostasis at endosomes

• Project/Area Number: 26440070
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Shizuoka University
• Principal Investigator: Kimura Yoko 静岡大学, 農学部, 教授 (80291152)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: ユビキチン / ホメオスタシス / エンドソーム / タンパク質分解 / 脱ユビキチン化酵素 / Rfu1 / Bro1 / 熱ストレス / Vドメイン / Alix / Ｒｉｍ２０ / 酵母 / V ドメイン

Outline of Final Research Achievements

Yeast Rfu1 (Regulator for free ubiquitin chain 1) localizes to endosomes and plays a role in ubiquitin homeostasis by inhibiting the activity of Doa4.We showed that Bro1, a class E vacuolar protein sorting protein that recruits Doa4 to endosomes, also recruits Rfu1 to endosomes by the direct interaction between a region containing the YPEL motif in Rfu1 and the V domain in Bro1, which could be analogous to the interaction between the mammalian Alix V domain and YPXnL motifs of target proteins. Overexpression of Bro1 and the V domain prevented Rfu1 degradation after heat shock. Rfu1 degradation partly involved the proteasome and Rsp5.Finally, we showed that Rim20, another yeast Bro1 family protein, binds to Rim101 through the interaction between the V domain and the YPIKL motif. These results suggest that interactions between V domains and YP(X)nL motifs are conserved from yeast to mammalian cells. We also showed the specificities of each V domain to their target protein.

Research Products

• [Journal Article] Conserved Mode of Interaction between Yeast Bro1 Family V Domains and YP(X)nL Motif-Containing Target Proteins (2015)
  • Author(s): Yoko Kimura, Mirai Tanigawa, Junko Kawawaki, Kenji Takagi, Tsunehiro Mizushima, Tatsuya Maeda, Keiji Tanaka
  • Journal Title: Eukaryotic Cell Volume: 14 Issue: 10 Pages: 976-982
  • DOI: 10.1128/ec.00091-15
  • NAID: 120005671359
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] VCP/Cdc48 rescues the growth defect of a GPI10 mutant in yeast (2015)
  • Author(s): Ohnuma Y, Takata T, Kawawaki J, Yasuda K, Tanaka K, Kimura Y, Kakizuka A.
  • Journal Title: FEBS Lett. Volume: 589 Issue: 5 Pages: 576
  • DOI: 10.1016/j.febslet.2015.01.017
  • NAID: 120005531440
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The ESCRT-III adaptor protein Brol controls functions of regulator for free ubiauitin chains 1 (Rful) in ubiquitin homeostasis (2014)
  • Author(s): Kimura Y, Kawawaki J, Kakiyama Y, Shimoda A, and Tanaka K
  • Journal Title: J Biol Chem Volume: 289 Issue: 31 Pages: 21760-21769
  • DOI: 10.1074/jbc.m114.550871
  • Peer Reviewed
• [Journal Article] Ubiquitin is phosphorylated by PINK1 to activate parkin. (2014)
  • Author(s): Koyano, F., Okatsu, K., Kosako, H., Tamura, Y., Go, E., Kimura, M., Kimura, Y., Tsuchiya, H., Yoshihara, H., Hirokawa, T., Endo, T., Fon. E. A., Trempe, J. F., Saeki, Y., Tanaka, K., and Matsuda, N.
  • Journal Title: Nature Volume: 510(in press) Issue: 7503 Pages: 162-166
  • DOI: 10.1038/nature13392
  • Peer Reviewed
• [Presentation] Analysis of interaction between yeast Bro1 family V domains and YP(X)nL motif-containing target proteins (2016)
  • Author(s): Yoko Kimura
  • Organizer: 14th International Congress on Yeasts
  • Place of Presentation: Awaji Yumebudai, Awaji, Hyogo, Japan
  • Year and Date: 2016-09-11
  • Int'l Joint Research
• [Presentation] Conserved Mode of Interaction between Yeast Bro1 Family V Domains and YP(X)nL Motif-Containing Target Proteins (2015)
  • Author(s): Yoko Kimura, Mirai Tanigawa, Junko Kawawaki, Kenji Takagi, Tsunehiro Mizushima, Tatsuya Maeda, Keiji Tanaka
  • Organizer: BMB2015
  • Place of Presentation: 神戸ポートアイランド（神戸市中央区）
  • Year and Date: 2015-12-01
• [Presentation] 酵母Bro1およびRim20のV ドメインの解析 (2015)
  • Author(s): 木村 洋子、谷川美頼、川脇純子、高木賢治、水島恒裕、前田達哉・田中啓二
  • Organizer: 酵母遺伝学フォーラム
  • Place of Presentation: 広島大学〈広島県東広島市）
  • Year and Date: 2015-08-31
• [Presentation] エンドソーム関連タンパク質Bro1とユビキチンの量的制御因子Rfu1の相互作用の解析 (2014)
  • Author(s): A1木村洋子.川脇純子.田中啓二
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜・横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] VCP/Cdc48過剰発現スクリーニングから得られたgpi10変異体の解析 (2014)
  • Author(s): 大沼洋平・木村洋子・川脇純子・田中啓二・垣塚彰
  • Organizer: 酵母遺伝学フォーラム
  • Place of Presentation: 東京大学・東京
  • Year and Date: 2014-09-01 – 2014-09-03
• [Remarks] 静岡大学 木村研究室
  • URL: http://kimurapqchs.agr.shizuoka.ac.jp/
• [Remarks] 静岡大学 木村研究室
  • URL: http://kimurapqchs.agr.shizuoka.ac.jp/wordpress/