Codon selection mechanism and protein translocation mechanism of a calcium channel regulatory subunit in budding yeast

• Project/Area Number: 19K06539
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43030:Functional biochemistry-related
• Research Institution: Tokyo Gakugei University
• Principal Investigator: IIDA Hidetoshi 東京学芸大学, 教育学部, 名誉教授 (70124435)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2022: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
• Keywords: 細胞内タンパク質輸送機構 / Nグリコシル化 / シグナルペプチド / カルシウムチャネル / 出芽酵母 / 小胞体 / 膜輸送機構 / Nグリコシレーション / 翻訳開始 / 開始コドン / Nグルコシレーション / 翻訳開始コドン / 翻訳 / 膜輸送

Outline of Research at the Start

Mid1は、酵母が種々のストレスを受けた時に生命を守るためにカルシウムシグナルを発生するカルシウムチャネルの制御サブユニットであり、本研究代表者が発見し、当該分野をリードしつつ長年研究したタンパク質である。本研究では、Mid1の膜輸送において、少なくとも２つのシグナルペプチド（SP）がはたらくことを明らかにし、酵母のストレス応答と関連づけてその２つを使い分けるメカニズムを明らかにしようと計画されている。このような研究はタンパク質生合成の分野で他に例がなく、学術的に重要である。この研究が成し遂げられた暁には、広く生物がもつストレス応答機構に新たな１ページを加えることができ、創造性が極めて高い。

Outline of Final Research Achievements

The aim of this study is to elucidate the mechanisms of translation-initiation-codon selection and membrane trafficking using Mid1, a regulatory subunit of a calcium channel in yeast. Mid1 has an N-terminal signal sequence (N-SP) to enter the endoplasmic reticulum lumen. We have obtained data suggesting that Mid1 has a C-terminal signal peptide (C-SP) as well as the N-SP. In this study, we investigated (1) the amino acid sequence of C-SP and (2) under what conditions the C-SP is used. In (1), we were close to the final clarification of it, while in (2), we are still in an early stage and need further research.

Academic Significance and Societal Importance of the Research Achievements

細胞質で合成されたタンパク質が細胞膜上または細胞外に輸送されるには、教科書的には、そのタンパク質のN末端にシグナルペプチドと呼ばれる約20個のアミノ酸残基からなる領域（N-SP）が必要である。酵母のMid1タンパク質はそのN-SPが無くても細胞膜まで到達できることを、我々は発見した。さらに、その到達ができる理由はMid1のC末端に第2のシグナルペプチド（C-SP）と呼ぶべき領域があることも発見した。本研究によりその領域を厳密に決めることができ、どのような生理条件の時にC-SPが使われるのかを明らかにできれば、タンパク質の細胞内輸送の制御機構に全く新しい概念を打ち立てることができる。

Research Products

• [Int'l Joint Research] Royal Botanic Garden Edinburgh, Scotland(英国)
• [Journal Article] Entanglement of Arabidopsis seedlings to a mesh substrate under microgravity conditions in KIBO on the ISS. (2022)
  • Author(s): Nakano, M., Furuichi, T., Sokabe, M., Iida, H., Yano, S., and Tatsumi, H.
  • Journal Title: Plants Volume: 11 Issue: 7 Pages: 956-956
  • DOI: 10.3390/plants11070956
  • Peer Reviewed / Open Access
• [Journal Article] MCAs in Arabidopsis are Ca2+-permeable mechanosensitive channels inherently sensitive to membrane tension. (2021)
  • Author(s): Yoshimura, K., Iida, K., and Iida, H.
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 6074-6074
  • DOI: 10.1038/s41467-021-26363-z
  • Peer Reviewed / Open Access
• [Journal Article] Mix and match: Patchwork domain evolution of the land plant-specific Ca2+-permeable mechanosensitive channel MCA. (2021)
  • Author(s): Nishii, K., Moeller, M., and Iida, H.
  • Journal Title: PLoS ONE Volume: 16
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A Method Enabling Comprehensive Isolation of Arabidopsis Mutants Exhibiting Unusual Root Mechanical Behavior (2021)
  • Author(s): Tojo, H., Nakamura, A., Ferjani, A., Kazama, Y., Abe, T., and Iida, H.
  • Journal Title: Frontiers in Plant Science Volume: - Pages: 1-8
  • DOI: 10.3389/fpls.2021.646404
  • Peer Reviewed / Open Access
• [Journal Article] The root growth reduction in response to mechanical stress involves ethylene-mediated microtubule reorganization and transmembrane receptor-mediated signal transduction in Arabidopsis (2021)
  • Author(s): Okamoto, T., Takatani, S., Motose, H., Iida, H., and Takahashi, T.
  • Journal Title: Plant Cell Reports Volume: 40 Issue: 3 Pages: 575-582
  • DOI: 10.1007/s00299-020-02653-6
  • Peer Reviewed / Open Access
• [Journal Article] The gravistimulation-induced very slow Ca 2+ increase in Arabidopsis seedlings requires MCA1, a Ca 2+-permeable mechanosensitive channel (2021)
  • Author(s): Nakano, M,m Furuichi, T., Sokabe, M., Iida, H., and Tatsumi H.
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 1-11
  • DOI: 10.1038/s41598-020-80733-z
  • Peer Reviewed / Open Access
• [Journal Article] Mix and match: Patchwork domain evolution of the land plant-specific Ca2+-permeable mechanosensitive channel MCA (2021)
  • Author(s): Nishii, K., Moeller, M., and Iida, H.
  • Journal Title: PLoS ONE Volume: 16 Issue: 4 Pages: 1-20
  • DOI: 10.1371/journal.pone.0249735
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Highly conserved extracellular residues mediate interactions between pore-forming and regulatory subunits of the yeast Ca2+ channel related to the animal VGCC/NALCN family (2020)
  • Author(s): Hayashi, T., Oishi K., Kimura, M., Iida, K., and Iida, H.
  • Journal Title: Journal of Biological Chemistry Volume: 295 Issue: 37 Pages: 13008-13022
  • DOI: 10.1074/jbc.ra120.014378
  • Peer Reviewed / Open Access
• [Journal Article] The ER-associated protease Ste24 prevents N-terminal signal peptide?independent translocation into the ER in Saccharomyces cerevisiae (2020)
  • Author(s): Hosomi Akira、Iida Kazuko、Cho Toshihiko、Iida Hidetoshi、Kaneko Masashi、Suzuki Tadashi
  • Journal Title: Journal of Biological Chemistry Volume: in press Issue: 30 Pages: 10406-10419
  • DOI: 10.1074/jbc.ra120.012575
  • Peer Reviewed / Open Access
• [Presentation] MCAs are land plant-specific, inherently mechanosensitive Ca2+-permeable channels (2022)
  • Author(s): Hidetoshi Iida, Kazuko Iida, Kanae Nishii, Michael Moeller, and Kenjiro Yoshimura
  • Organizer: 10th International Plant Biomechanics Conference
  • Int'l Joint Research
• [Presentation] 植物のメカノセンサー (2022)
  • Author(s): 飯田秀利
  • Organizer: 第45回日本分子生物学会年会ワークショップ「植物メカノバイオロジー：一細胞レベルからから個体レベルへ
  • Invited
• [Presentation] Ca2+-permeable mechanosensitive channels MCA1 and 2: their molecular nature and physiological roles (2022)
  • Author(s): Hidetoshi Iida
  • Organizer: 第86回日本植物学会大会JPRシンポジウム「Mechanical Forces in Plant Growth and Development」
  • Int'l Joint Research / Invited
• [Presentation] 出芽酵母のカルシウムチャネル制御サブユニットMid1の膜輸送に重要な第２の領域 (2021)
  • Author(s): 飯田和子、飯田秀利
  • Organizer: 日本分子生物学会
• [Presentation] Plant mechanosensitive channels MCA1 and MCA2: their structure, function, and roles (2019)
  • Author(s): Iida, H.
  • Organizer: Department of Biology, Norwegian University of Science and Technology. Trondheim, Norway
  • Invited
• [Presentation] Molecular mechanisms of plant growth and developments in the ISS, a closed environment under microgravity (2019)
  • Author(s): Furuichi, T., Nakano, M., Matsunami, S., Kato, A., Nakazawa, E., Nagano, Y., Fujita, H., Iida, H., and Tatsumi H.
  • Organizer: The 40th Annual Meeting of the International Society for Gravitational Physiology and Space Life Science and Medicine, Nagoya
  • Int'l Joint Research
• [Presentation] Identification of extracellular residues that anchor the interaction between the pore-forming subunit Cch1 and the essential regulatory subunit Mid1 of a yeast Ca2+ channel (2019)
  • Author(s): Iida, H., Hayashi, T., Oishi, K., Kimura, M., and Iida, K.
  • Organizer: The 29th International Conference on Yeast Genetics and Molecular Biology, The Swedish Exhibition & Congress Centre, Gothenburg, Sweden
  • Int'l Joint Research
• [Presentation] N末端シグナルペプチドを欠失した膜タンパク質のERへのトランスロケーション機構 (2019)
  • Author(s): 飯田和子、飯田秀利
  • Organizer: 第52回酵母遺伝学フォーラム、静岡市清水文化会館マリナート（清水市）、静岡
• [Presentation] In vitroで合成・精製したシロイヌナズCa2+透過性機械受容チャネルのイオン透過性 (2019)
  • Author(s): 飯田秀利、飯田和子、吉村建二郎
  • Organizer: 日本植物学会第83回大会、東北大学川内北キャンパス（仙台市）
• [Presentation] 苔類ゼニゴケMCA1 オルソログの成長・生殖過程における機能解析 (2019)
  • Author(s): 岩野 惠、末次憲之、西浜竜一、石田咲子、木村 緑、飯田和子、飯田秀利、永井健冶、河内孝之
  • Organizer: 日本植物学会第83回大会、東北大学川内北キャンパス（仙台市）
• [Presentation] 植物に固有の機械受容チャネルの構造と機械刺激受容機構 (2019)
  • Author(s): 飯田秀利、飯田和子、吉村建二郎
  • Organizer: 第42回日本分子生物学会年会、福岡国際会議場
• [Book] 植物の機械感覚 in「生き物と音の辞典」（一社） 生物音響学会編 (2019)
  • Author(s): 飯田秀利
  • Total Pages: 441
  • Publisher: 朝倉書店
  • ISBN: 9784254171679