Development of a novel protein production system in mammalian cells.

• Project/Area Number: 25660295
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied molecular and cellular biology
• Research Institution: Kyoto University
• Principal Investigator: MASUDA Seiji 京都大学, 生命科学研究科, 准教授 (20260614)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: mRNA / タンパク質 / 輸送受容体 / 核外輸送

Outline of Final Research Achievements

The proteins for the clinical use are produced in mammalian cells because they needs post translational modification such as glycosylation, formation of adequate disulfide-bond, acylation and phosphorylation for their activity in human body. The protein production in mammalian cells, however, takes much cost than that in bacteria. The author found that the mRNA export process becomes the limited step than transcription or translation process. In the present study, mRNA export receptor is created to establish a novel protein production system in mammalian cells by modifying the sequence of mRNA export receptor. The mutant receptor produce more reporter protein than the wild type export receptor.

Research Products

• [Journal Article] RNA export pathways in eukaryotes. (2015)
  • Author(s): Okamura, M, Inose, H. and Masuda, S.
  • Journal Title: Genes Volume: 6 Issue: 1 Pages: 125-149
  • DOI: 10.3390/genes6010124
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Gene Regulation through mRNA Expression. (2015)
  • Author(s): Inose, H., Mukai, K., Ito, M. and Masuda, S.
  • Journal Title: Adv. Biol. Chem. Volume: 5 Issue: 02 Pages: 45-57
  • DOI: 10.4236/abc.2015.52005
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Erythropoietin receptor antagonist suppressed ectopic hemoglobin synthesis in xenografts of HeLa cells to promote their destruction. (2015)
  • Author(s): Yasuda, Y., Fujita, M., Koike, E., Obata, K., Shiota, M., Kotani, Y., Musha, T., Tsuji-Kawahara, S., Satou, T., Masuda, S, Okano, J., Yamasaki, H., Okumoto, K., Uesugi, T., Nakao, S.,Hoshiai, H. and Mandai, M.
  • Journal Title: PLoS ONE Volume: 10 Issue: 4 Pages: e0122458-e0122458
  • DOI: 10.1371/journal.pone.0122458
  • Peer Reviewed / Open Access
• [Journal Article] Cooperative functions of ZnT1, metallothionein and ZnT4 in the cytoplasm are required for full (2013)
  • Author(s): Fujimoto S, Kimura T, Nagao M and Kambe T, et al. （11人中11番目）
  • Journal Title: PLOS ONE Volume: 8 Issue: 10 Pages: e77445-e77445
  • DOI: 10.1371/journal.pone.0077445
  • Peer Reviewed
• [Journal Article] Identification of 6-octadecynoic acid from a methanol extract of Marrubium vulgare L. as a peroxisome proliferator-activated receptor γ agonist (2013)
  • Author(s): Ohtera, A., Miyamae, Y., Nakai, N., Kawachi, A., Kawada, K., Han, J., Isoda, H., Neffati, M., Akita, T., Maejima, K., Masuda, S., Kambe, T., Mori, N., Irie, K. and Nagao, M
  • Journal Title: Biochem. Biophys. Res. Commun Volume: 440 Issue: 2 Pages: 204-209
  • DOI: 10.1016/j.bbrc.2013.09.003
  • Peer Reviewed
• [Journal Article] Trafficking of mRNAs containing ALREX-promoting elements through nuclear speckles (2013)
  • Author(s): Akef, A., Zhang, H., Masuda, S. and Palazzo, AF
  • Journal Title: Nucleus Volume: 4 Issue: 4 Pages: 326-40
  • DOI: 10.4161/nucl.26052
  • Peer Reviewed
• [Presentation] The identification of regions regulates different complex formations of UAP56 and URH49, which are essential for mRNA export. (2015)
  • Author(s): 藤田 賢一、伊藤 慶紗、増田 誠司
  • Organizer: The 13th International Student Seminar
  • Place of Presentation: 紫蘭会館、京都
  • Year and Date: 2015-03-03
• [Presentation] Involvement of nuclear factor CHERP in alternative splicing (2015)
  • Author(s): 向井琴美、増田誠司
  • Organizer: The 13th International Student Seminar
  • Place of Presentation: 紫蘭会館、京都
  • Year and Date: 2015-03-03
• [Presentation] The identification of regions regulates different complex formations of UAP56 and URH49, which are essential for mRNA export (2015)
  • Author(s): 藤田 賢一、伊藤 慶紗、増田 誠司
  • Organizer: KANSAI JSBBA 1st Student Forum
  • Place of Presentation: 洛友会館、京都
  • Year and Date: 2015-01-31
• [Presentation] The identification of regions regulates different complex formations of UAP56 and URH49, which are essential for mRNA export (2014)
  • Author(s): 藤田 賢一、伊藤 慶紗、増田 誠司
  • Organizer: Joint Australia and Japan RNA Meeting
  • Place of Presentation: シドニー、オーストラリア
  • Year and Date: 2014-11-04
• [Presentation] 核内因子CHERPの機能解析 (2014)
  • Author(s): 向井琴美、南裕基、永尾雅哉、神戸大朋、宮前友策、増田誠司
  • Organizer: 2014年度日本農芸化学会 関西支部大会
  • Place of Presentation: 奈良先端科学技術大学院大学、奈良
  • Year and Date: 2014-09-20
• [Presentation] 進化的観点からUAP56とURH49の複合体形成と選択的mRNA輸送能の分岐点を探る (2014)
  • Author(s): 藤田 賢一、伊藤 慶紗、増田 誠司
  • Organizer: 第 16 回日本 RNA 学会年会
  • Place of Presentation: ウインクあいち、名古屋
  • Year and Date: 2014-07-23 – 2014-07-24
• [Presentation] mRNA輸送にはDBP5の核側からの核膜孔配位が必要だ (2014)
  • Author(s): 志岐 拓哉、岡村 真純、増田 誠司
  • Organizer: 第 16 回日本 RNA 学会年会
  • Place of Presentation: ウインクあいち、名古屋
  • Year and Date: 2014-07-23 – 2014-07-24
• [Presentation] mRNA核外輸送因子UAP56ならびにURH49の複合体形成の違いを制御する蛋白質領域の同定 (2014)
  • Author(s): 藤田 賢一、伊藤 慶紗、宮前 友策、神戸 大朋、永尾 雅哉、増田 誠司
  • Organizer: 日本農芸化学会関西支部 第484回講演会
  • Place of Presentation: 京都府立大学、京都
  • Year and Date: 2014-05-24
• [Presentation] 動物培養細胞での組換え蛋白質高生産に向けた mRNAの効率的核外輸送機構の開発 (2014)
  • Author(s): 猪瀬 春子、松村 嘉員、宮前 友策、神戸 大朋、永尾 雅哉、増田 誠司
  • Organizer: 日本農芸化学会関西支部 第484回講演会
  • Place of Presentation: 京都府立大学、京都
  • Year and Date: 2014-05-24
• [Presentation] 動物細胞の核外輸送過程を利用したタンパク質生産法 (2014)
  • Author(s): 増田誠司
  • Organizer: 日本農芸化学会
  • Place of Presentation: 東京
  • Invited
• [Presentation] mRNA成熟阻害活性を指標とする抗ガン化合物の探索と産業利用 (2014)
  • Author(s): 増田誠司
  • Organizer: 日本農芸化学会
  • Place of Presentation: 東京
  • Invited
• [Presentation] he GLE1 mutation identified in LCCS1 affects nuclear mRNA export (2013)
  • Author(s): Masumi Okamura, Takuya Shiki, Seiji Masuda
  • Organizer: RiboClub
  • Place of Presentation: ケベック、カナダ
• [Presentation] 胎性致死の遺伝病LCCS1にて同定されたGLE1の変異FinMajorはmRNAの核外輸送効率を低下させる (2013)
  • Author(s): 岡村真純、志岐拓哉、増田誠司
  • Organizer: 日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] mRNA核外輸送因子UAP56ならびにURH49における複合体形成差異を決定する領域の同定
  • Author(s): 藤田賢一、南裕基、増田誠司
  • Organizer: 日本RNA学会
  • Place of Presentation: 愛媛
• [Presentation] The identification of regions regulates different complex formations of UAP56 and URH49, which are essential for mRNA export
  • Author(s): Kenichi Fujita, Yuki Minami, Seiji Masuda
  • Organizer: The 12th International Student Seminar
  • Place of Presentation: 京都
• [Book] 動物細胞の培養を成功させる条件設定集 (2014)
  • Author(s): 岡村真純、平山瑞季、猪瀬春子、増田誠司
  • Publisher: 技術情報協会
• [Remarks] http://www.bunshioutou.lif.kyoto-u.ac.jp
• [Remarks] 分子応答機構学研究室ホームページ
  • URL: http://www.lif.kyoto-u.ac.jp/labs/bunshioutou/
• [Remarks] 京都大学大学院生命科学研究科分子応答機構学
  • URL: http://www.lif.kyoto-u.ac.jp/j/modules/education/content0040.html