Structure-function analysis of Tup1-Cyc8 corepressor

• Project/Area Number: 25440022
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Ritsumeikan University (2015); Osaka University (2013-2014)
• Principal Investigator: Matsumura Hiroyoshi 立命館大学, 生命科学部, 教授 (30324809)
• Co-Investigator(Kenkyū-buntansha): MUKAI YUKIO 長浜バイオ大学, バイオサイエンス学部, 准教授 (60252615) ; SAHARA TAKEHIKO 産業技術総合研究所, 生物プロセス研究部門, 主任研究員 (40357166) ; MORITA NAOKI 産業技術総合研究所, 生物プロセス研究部門, 研究グループ長 (60371085)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: Tup1-Cyc8 / コリプレッサー / 構造と機能 / Tup1-Cyc8複合体 / 構造機能相関 / Tup-Cyc8複合体 / 構造機能解析

Outline of Final Research Achievements

The yeast Cyc8p-Tup1p protein complex is a general transcriptional corepressor of genes involved in many physiological processes. In this study, we analyze the structure and function of Cyc8p-Tup1p protein complex using X-ray crystallography and small angle X-ray scattering. Based on the three-dimensional structure, we mutated Tup1p and characterized the Cyc8p and Tup1p function. Furthermore, we succeeded to express and purify the full-length Tup1p using cold-shock yeast expression system.

Research Products

• [Journal Article] Epiregulin Recognition Mechanisms by Anti-epiregulin Antibody 9E5: STRUCTURAL, FUNCTIONAL, AND MOLECULAR DYNAMICS SIMULATION ANALYSES. (2016)
  • Author(s): Kado Y1, Mizohata E2, Nagatoishi S3, Iijima M4, Shinoda K4, Miyafusa T3, Nakayama T2, Yoshizumi T2, Sugiyama A4, Kawamura T4, Lee YH4, Matsumura H2, Doi H4, Fujitani H4, Kodama T4, Shibasaki Y4, Tsumoto K5, Inoue T6.
  • Journal Title: J Biol Chem. Volume: 291 Issue: 5 Pages: 2319-2330
  • DOI: 10.1074/jbc.m115.656009
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Supersensitive odor discrimination is controlled in part by initial transient interactions between the most sensitive dorsal olfactory receptors and G-proteins (2016)
  • Author(s): Sato T, Kobayakawa R, Kobayakawa K, Emura M, Itohara S, Kawasaki T, Tsuboi A, Matsumura H
  • Journal Title: Receptors & Clinical Investigation Volume: 3
  • DOI: 10.14800/rci.1117
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structure-based design and synthesis of a bivalent iminobiotin analog showing strong affinity toward a low immunogenic streptavidin mutant. (2015)
  • Author(s): Kawato T, Mizohata E, Shimizu Y, Meshizuka T, Yamamoto T, Takasu N, Matsuoka M, Matsumura H, Kodama T, Kanai M, Doi H, Inoue T, Sugiyama A
  • Journal Title: Biosci Biotechnol Biochem. Volume: 79 Issue: 4 Pages: 640-642
  • DOI: 10.1080/09168451.2014.991692
  • Peer Reviewed / Open Access
• [Journal Article] Yeast Cyc8p and Tup1p proteins function as coactivators for transcription of Stp1/2p-dependent amino acid transporter genes (2015)
  • Author(s): Naoki Tanaka, Yukio Mukai
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 468 Issue: 1-2 Pages: 32-38
  • DOI: 10.1016/j.bbrc.2015.11.001
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Structure-based design of a streptavidin mutant specific for an artificial biotin analogue. (2015)
  • Author(s): Kawato, T.; Mizohata, E.; Shimizu, Y.; Meshizuka, T.; Yamamoto, T. Takasu, N.; Matsuoka, M.; Matsumura, H.; Kodama, T.; Kanai, M.; Doi, H.; Inoue, T.; Sugiyama, A.
  • Journal Title: J. of Biochemistry Volume: 1 Issue: 6 Pages: 1-9
  • DOI: 10.1093/jb/mvv004
  • Peer Reviewed / Open Access
• [Journal Article] Selective crystallization of metastable phase of acetaminophen by ultrasonic irradiation (2015)
  • Author(s): Y. Mori, M. Maruyama, Y. Takahashi, K. Ikeda, S. Fukukita, H. Y. Yoshikawa, S. Okada, H. Adachi, S. Sugiyama, K. Takano, S. Murakami, H. Matsumura, T. Inoue, M. Yoshimura, and Y. Mori
  • Journal Title: Applied Physics Express Volume: Vol.8, No.6 Issue: 6 Pages: 065501-065501
  • DOI: 10.7567/apex.8.065501
  • NAID: 210000137563
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Development of protein seed crystals reinforced by high-strength hydrogels (2015)
  • Author(s): Shigeru Sugiyama, Noriko Shimizu, Keisuke Kakinouchi, Osamu Hiraoka, Hiroyoshi Matsumura, Hiroshi Y. Yoshikawa, Yoshinori Takahashi, Mihoko Maruyama, Masashi Yoshimura, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Michio Murata, and Yusuke Mori
  • Journal Title: CrystEngComm Volume: 17 Issue: 42 Pages: 8064-8071
  • DOI: 10.1039/c5ce00844a
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Insights into unknown foreign ligand in copper nitrite reductase (2015)
  • Author(s): Yohta Fukuda, Tse Ka Man, Yuji Kado, Eiichi Mizohata, Hiroyoshi Matsumura, Tsuyoshi Inoue
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 464 Pages: 622-628
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The N-terminal acidic residue of the cytosolic helix 8 of an odorant receptor is responsible for different response dynamics via G-protein (2015)
  • Author(s): T. Kawasaki, T. Saka, S. Mine, E. Mizohata, T. Inoue, H. Matsumura, T. Sato
  • Journal Title: FEBS Lett. Volume: 589 Issue: 10 Pages: 1136-1143
  • DOI: 10.1016/j.febslet.2015.03.025
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Protein Pept. Lett. (2015)
  • Author(s): R.N. Abd Rahman, M.S. Ali, S. Sugiyama, A.T. Leow, T. Inoue, M. Basri, A.B. Salleh, and H. Matsumura
  • Journal Title: A comparative analysis of microgravity and earth grown thermostable T1 Lipase crystals using HDPCG apparatus Volume: 22 Pages: 173-179
  • Peer Reviewed
• [Journal Article] Crystal structure of FtsA from Staphylococcus aureus (2014)
  • Author(s): J. Fujita, Y. Maeda, C. Nagao, Y. Tsuchiya, Y. Miyazaki, M. Hirose, E. Mizohata, Y. Matsumoto, T. Inoue, K. Mizuguchi, and H. Matsumura
  • Journal Title: FEBS Lett. Volume: 588 Issue: 10 Pages: 1879-1885
  • DOI: 10.1016/j.febslet.2014.04.008
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Structural basis for the recognition-evasion arms race between Tomato mosaic virus and the resistance gene Tm-1 (2014)
  • Author(s): K. Ishibashi, Y. Kezuka, C. Kobayashi, M. Kato, T. Inoue, T. Nonaka, M. Ishikawa, H. Matsumura, and E. Katoh
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 111 Issue: 33
  • DOI: 10.1073/pnas.1407888111
  • Peer Reviewed
• [Journal Article] 転写制御因子Tup1-N 末端ドメインのX線結晶構造とその複合体形成メカニズム (2013)
  • Author(s): 中村太一、松村浩由、井上豪、日下菜花、田中直子、向由起夫
  • Journal Title: 日本結晶学会誌 Volume: 55 Pages: 110-115
  • NAID: 10031168499
  • Peer Reviewed
• [Journal Article] Expression, purification, crystallization and preliminary crystallographic study of FtsA from methicillin-resistant Staphylococcus aureus (2013)
  • Author(s): J. Fujita, Y. Miyazaki, M. Hirose, C. Nagao, E. Mizohata, Y. Matsumoto, K. Mizuguchi, T. Inoue, and H. Matsumura
  • Journal Title: Acta. Cryst. Volume: F69 Issue: 8 Pages: 895-898
  • DOI: 10.1107/s1744309113017727
  • Peer Reviewed
• [Presentation] 離合集散する柔らかいタンパク質の構造機能解析 (2016)
  • Author(s): 松村浩由
  • Organizer: 蛋白研セミナー
  • Place of Presentation: 東京大学先端科学技術研究センター（東京都、目黒区）
  • Year and Date: 2016-03-01
  • Invited
• [Presentation] 出芽酵母コリプレッサーTup1pのヒストン結合領域がもつ新たな役割 (2015)
  • Author(s): 田中 直子、向 由起夫
  • Organizer: 日本分子生物学会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
• [Presentation] コリプレッサーTup1pの4-へリックスバンドル領域における3連続グ ルタミン酸の機能解析 (2015)
  • Author(s): 竹下 将平、田中 直子、向 由起夫
  • Organizer: Yeast Workshop
  • Place of Presentation: せとうち児島ホテル（岡山県倉敷市）
  • Year and Date: 2015-11-13
• [Presentation] 黄色ブドウ球菌由来細胞分裂タンパク質FtsZの構造変化機構 (2015)
  • Author(s): 藤田純三、前田陽子、齋藤有紀、溝端栄一、井上豪、松村浩由
  • Organizer: 日本結晶学会年会
  • Place of Presentation: 大阪府立大学（大阪府堺市）
  • Year and Date: 2015-10-17
• [Presentation] 出芽酵母コリプレッサーTup1によるヒストン結合を必要としない転写抑制機構 (2014)
  • Author(s): 田中直子、布施智博、清水光弘、向由起夫
  • Organizer: 日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-26
• [Presentation] 出芽酵母コリプレッサーTup1の4へリックスバンドル領域における3連続グルタミン酸の機能解析 (2014)
  • Author(s): 竹下将平、田中直子、向由起夫
  • Organizer: 分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-26
• [Presentation] コリプレッサーTup1pの4-へリックスバンドル領域における3連続グルタミン酸の機能解析 (2014)
  • Author(s): 竹下将平、向由起夫
  • Organizer: Yeast Workshop
  • Place of Presentation: ビューポートくれ（広島県呉市）
  • Year and Date: 2014-11-14
• [Presentation] MRSA由来細胞分裂タンパク質FtsAの構造―機能相関解析 (2014)
  • Author(s): 藤田純三、前田陽子、齋藤有紀、清水伸隆、野田勝紀、長尾知生子、土屋裕子、溝端栄一、内山進、松本佳巳、水口賢司、井上豪、松村浩由
  • Organizer: 日本結晶学会
  • Place of Presentation: 東京大学農学部（東京都文京区）
  • Year and Date: 2014-11-01 – 2014-11-02
• [Presentation] 出芽酵母コリプレッサー複合体Cyc8-Tup1はトリプトファン取込み酵素遺伝子TAT1とTAT2の転写活性化に必要である (2014)
  • Author(s): 田中直子、田井晶子、向由起夫
  • Organizer: 日本遺伝学会
  • Place of Presentation: 長浜バイオ大学（滋賀県長浜市）
  • Year and Date: 2014-09-19
• [Presentation] ヒストン結合領域を欠失したコリプレッサーTup1の機能解析 (2014)
  • Author(s): 田中直子、布施智博、清水光弘、向由起夫
  • Organizer: 酵母遺伝学フォーラム
  • Place of Presentation: 東京大学弥生講堂（東京都文京区）
  • Year and Date: 2014-09-01 – 2014-09-02
• [Presentation] MRSA由来細胞分裂タンパク質FtsAのX線結晶構造解析 (2014)
  • Author(s): 藤田純三、前田陽子、宮﨑祐満、廣瀬未果、溝端栄一、井上豪、松村浩由
  • Organizer: 日本蛋白質科学会
  • Place of Presentation: ワークピア横浜 / 横浜産貿ホール マリネリア（神奈川県横浜市）
  • Year and Date: 2014-06-25 – 2014-06-27
• [Presentation] CP12-GAPDH-PRK複合体形成によるカルビン回路調節機構の解明 (2014)
  • Author(s): 松村浩由、清水伸隆、木津奈都子、吉田将康、荒武将弘、溝端栄一、田茂井政宏、和田野晃、重岡成、井上豪
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 東京
• [Presentation] Structural basis for assembly of the CP12-GAPDH-PRK complex (2014)
  • Author(s): Hiroyoshi Matsumura, Nobutaka Shimizu, Natsuko Kizu, Masayasu Yoshida, Masahiro Aratake, Eiichi Mizohata, Masahiro Tamoi, Akira Wadano, Sshigeru Shigeoka, Tsuyoshi Inoue
  • Organizer: 日本農芸化学会2014年度大会
  • Place of Presentation: 富山
• [Presentation] カルビン回路調節システムの構造基盤 (2013)
  • Author(s): 松村浩由
  • Organizer: アグリバイオシンポジウム 2013
  • Place of Presentation: 奈良
  • Invited
• [Presentation] SAXS and XRD analyses of the Calvin cycle regulatory complexes (2013)
  • Author(s): Hiroyoshi Matsumura, Nobutaka Shimizu Tsuyoshi Inoue
  • Organizer: 日本生物物理学会年会
  • Place of Presentation: 京都
  • Invited
• [Presentation] 結晶解析と溶液散乱による相関解析 (2013)
  • Author(s): 松村 浩由
  • Organizer: タンパク質X線溶液散乱講習会
  • Place of Presentation: 筑波
  • Invited
• [Presentation] Candida albicans Tup1コリプレッサーの機能解析 (2013)
  • Author(s): 田中 直子、向 由起夫
  • Organizer: 酵母遺伝学フォーラム
  • Place of Presentation: 仙台
• [Presentation] 酵母コリプレッサーTup1ヒストン結合領域の機能解析 (2013)
  • Author(s): 田中直子、向由起夫
  • Organizer: 日本分子生物学会
  • Place of Presentation: 神戸