Substrate-induced degradation of yeast amino acid permeases and the autodegradation-signaling associated with dynamic structural changes

• Project/Area Number: 18K05397
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 38020:Applied microbiology-related
• Research Institution: Aoyama Gakuin University
• Principal Investigator: Abe Fumiyoshi 青山学院大学, 理工学部, 教授 (30360746)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 酵母アミノ酸輸送体 / 動的構造変化 / 自己分解シグナル / 出芽酵母 / トリプトファン輸送体 / ユビキチン分解 / 基質輸送 / アイソソーム / アミノ酸輸送体 / トリプトファン輸送体Tat2 / Eisosome / ユビキチン依存性分解 / 高圧増殖 / Ehg1/May24 / 栄養源センシング / ユビキチン化

Outline of Final Research Achievements

Nutrient starvation is a life-threatening stress while nutrient excess is also toxic in microorganisms. Among two yeast tryptophan permeases, Tat1 (the low-affinity type) was stably expressed in the plasma membrane whereas Tat2 (the high-affinity type) was rapidly degraded upon substrate addition. The D74R mutation in the acidic patch in the cytoplasmic domain abolished the substrate-induced Tat2 degradation. We hypothesize that the dynamic structural changes associated the substrate imports transmit the autodegradation signal of Tat2 allowing its own ubiquitination. We also explored cellular responses to nutrient limitation from aspects of poorly-characterized proteins and TORC1.

Academic Significance and Societal Importance of the Research Achievements

トリプトファンの動態は、細胞内NAD+の調節、トリプトフォールやセロトニンなど細胞間伝達物質の合成、ヒトではトリプトファン輸送体MCT10が甲状腺ホルモンの輸送を担うなど、他のアミノ酸にはないユニークさがある。またトリプトファンは希少なアミノ酸であるうえ、ヒトの必須アミノ酸でもある。よって低濃度トリプトファン条件下における取り込みと、それとは逆に基質過剰時の取り込み抑制が特に重要だ。本研究で得られた成果は、酵母のトリプトファン輸送体Tat2が広範な生物種におけるトリプトファン制御のモデルとなることを示すとともに、１タンパク質分子がシグナルの受容と伝達を同時に担うユニークな側面を映し出している。

Research Products

• [Journal Article] Amino acid homeostatic control by TORC1 in Saccharomyces cerevisiae under high hydrostatic pressure (2020)
  • Author(s): S. Uemura, T. Mochizuki, K. Amemiya, G. Kurosaka, M. Yazawa, K. Nakamoto, Y. Ishikawa, S. Izawa, F. Abe
  • Journal Title: J. Cell Sci. Volume: 133 Issue: 17
  • DOI: 10.1242/jcs.245555
  • Peer Reviewed
• [Journal Article] Draft Genome Sequence of Naganishia liquefaciens Strain N6, Isolated from the Japan Trench (2020)
  • Author(s): Han Yong-Woon、Kajitani Rei、Morimoto Hiroya、Palihati Maierdan、Kurokawa Yumiko、Ryusui Rie、Argunhan Bilge、Tsubouchi Hideo、Abe Fumiyoshi、Kajiwara Susumu、Iwasaki Hiroshi、Itoh Takehiko
  • Journal Title: Microbiology Resource Announcements Volume: 9 Issue: 47
  • DOI: 10.1128/mra.00827-20
  • NAID: 120007124437
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Over-expression of Caj1, a plasma membrane associated J-domain protein in Saccharomyces cerevisiae stabilizes amino acid permeases. (2020)
  • Author(s): Dobriyala, N., Sagarikaa, P., Shrivastavaa, A., Vermaa, A.K., Islama, Z., Guptaa, P. , Mochizuki, T., Abe, F., Sahi, C.
  • Journal Title: Biochim. Biophys. Acta Volume: 1862 Issue: 11 Pages: 183435-183435
  • DOI: 10.1016/j.bbamem.2020.183435
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A novel ER membrane protein Ehg1/May24 plays a critical role in maintaining multiple nutrient permeases in yeast under high-pressure perturbation. (2019)
  • Author(s): 1. Kurosaka, G., Uemura, S., Mochizuki, T., Kozaki, Y., Hozumi, A., Suwa, S., Ishii, R., Kato, Y., Imura, S., Ishida, N., Noda, Y., and Abe, F
  • Journal Title: Sci. Rep. Volume: 9 Issue: 1 Pages: 18341-18341
  • DOI: 10.1038/s41598-019-54925-1
  • Peer Reviewed / Open Access
• [Journal Article] Hyperactive mutation occurs adjacent to the essential glutamate 286 for transport in the yeast tryptophan permease Tat2 (2019)
  • Author(s): Amano Kaori、Ishii Ryoga、Mochizuki Takahiro、Takatsu Shiori、Abe Fumiyoshi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 509 Issue: 4 Pages: 1047-1052
  • DOI: 10.1016/j.bbrc.2019.01.038
  • Peer Reviewed
• [Journal Article] Identification of a regulatory element for yeast tryptophan permease Tat2 ubiquitination using high hydrostatic pressure. (2019)
  • Author(s): Ishii, R. , Aramaki, M., Mochizuki, T., and Abe, F
  • Journal Title: High Pressure Research Volume: in press
  • Peer Reviewed
• [Journal Article] Simulated microgravity triggers characteristic morphology and stress response in Saccharomyces cerevisiae (2018)
  • Author(s): Nemoto Shota、Ohnuki Shinsuke、Abe Fumiyoshi、Ohya Yoshikazu
  • Journal Title: Yeast Volume: 36 Issue: 2 Pages: 85-97
  • DOI: 10.1002/yea.3361
  • Peer Reviewed
• [Journal Article] The YPR153W gene is essential for the pressure tolerance of tryptophan permease Tat2 in the yeast Saccharomyces cerevisiae (2018)
  • Author(s): Kurosaka, G. and Abe, F.
  • Journal Title: High Pressure Research Volume: 30 Pages: 90-98
  • Peer Reviewed
• [Presentation] 高水圧によるTORC1の活性化と細胞内 アミノ酸ホメオスタシスにおける意義 (2020)
  • Author(s): 阿部文快、上村聡志、望月貴博、雨宮賢吾、中本景子、石川優、井沢真吾
  • Organizer: 酵母遺伝学フォーラム
• [Presentation] 新規小胞体膜タンパク質Ehg1による酵母栄養源輸送体の制御 (2019)
  • Author(s): 加藤祐介、黒坂豪祐、上村聡志、望月貴博、伊村咲希、石田夏穂、石井凌賀、野田陽一、阿部文快
  • Organizer: 酵母遺伝学フォーラム
• [Presentation] 出芽酵母の圧力適応に重要なシグナル伝達経路の探索 (2019)
  • Author(s): 望月貴博、波多野絵梨、藤山未奈、阿部文快
  • Organizer: 酵母遺伝学フォーラム
• [Presentation] 深海好圧性細菌Shewanella violaceaにおけるペプチド輸送体の大腸菌内発現と機能解析 (2019)
  • Author(s): 根來将吾、翠川雄斗、阿部文快
  • Organizer: 極限環境生物学会
• [Presentation] Functional linkage between poorly characterized genes and nutrient transport in yeast under high pressure (2018)
  • Author(s): Goyu Kurosaka, Satoshi Uemura, Takahiro Mochizuki, Yuri Kozaki, Akiko, Hozumi, Sayuri Suwa and Fumiyoshi Abe
  • Organizer: 10th International Conference on High Pressure Bioscience and Biotechnology (HPBB2018)
  • Int'l Joint Research
• [Presentation] Exploration of high-pressure sensor proteins in the yeast Saccharomyces cerevisiae (2018)
  • Author(s): Takahiro Mochizuki and Fumiyoshi Abe
  • Organizer: 10th International Conference on High Pressure Bioscience and Biotechnology (HPBB2018)
  • Int'l Joint Research
• [Presentation] A novel membrane protein Ehg1 physically interacts with nutrient permeases in the ER allowing yeast cells to grow under high pressure. (2018)
  • Author(s): Yusuke Kato, Saki Imura, Natsuho Ishida, Ryoga Ishi, Goyu Kurosaka, Takahiro Mochizuki, Satoshi Uemura and Fumiyoshi Abe
  • Organizer: 10th International Conference on High Pressure Bioscience and Biotechnology (HPBB2018)
  • Int'l Joint Research
• [Presentation] Activation of the EGO complex-TORC1 signaling pathway enables the yeast Saccharomyces cerevisiae cells to grow under high pressure (2018)
  • Author(s): Satoshi Uemura, Kengo Amemiya, Takahiro Mochizuki, Goyu Kurosaka, Miho Yazawa, and Fumiyoshi Abe
  • Organizer: 10th International Conference on High Pressure Bioscience and Biotechnology (HPBB2018)
  • Int'l Joint Research
• [Presentation] Heterologous expression and characterization of a peptide transporter from deep-sea piezophile Shewanella violacea in Escherichia coli (2018)
  • Author(s): Shogo Negoro, Yuto Midorikawa, Chiaki Kato and Fumiyoshi Abe
  • Organizer: 10th International Conference on High Pressure Bioscience and Biotechnology (HPBB2018)
  • Int'l Joint Research