Analysis of mRNA export mechanism and applied mRNA biotechnology in eukaryote.

• Project/Area Number: 26292053
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Kyoto University
• Principal Investigator: Masada Seiji 京都大学, 生命科学研究科, 准教授 (20260614)
• Co-Investigator(Kenkyū-buntansha): 井沢 真吾 京都工芸繊維大学, 工芸科学研究科, 准教授 (10273517)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,640,000 (Direct Cost: ¥12,800,000、Indirect Cost: ¥3,840,000); Fiscal Year 2016: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000)
• Keywords: mRNA export / UAP56 / URH49 / structure / complex formation / mRNA輸送 / TREX / AREX / 結晶 / mRNA / 核外輸送 / タンパク質生産 / mRNA輸送体 / タンパク質 / 複合体

Outline of Final Research Achievements

UAP56, a DEAD box RNA helicase, functions to mRNA export from the nucleus to the cytoplasm. In higher eukaryote, there is a paralog of UAP56, named URH49. Although their amino acid homology is more than 90%, they form different complex and export specific set of mRNAs. In this study, I analyzed the region to regulate the complex formation using molecular biology technique and the structural basis using　structural biology method. The results indicate that the region regulating the complex formation is within N domain of UAP56 and URH49 and that their structures are similar but the coordination of N and C domains were different between UAP56 and URH49 and the structure of linker region between N and C domains was also different.

Research Products

• [Journal Article] The isoflavone fraction from soybean presents mRNA maturation inhibition activity (2017)
  • Author(s): Kurata M, Murata Y, Momma K, Fouad Ali Mursi I, Takahashi M, Miyamae Y, Kambe T, Nagao M, Narita H, Shibuya Y, Masuda S
  • Journal Title: Biosci Biotechnol Biochem Volume: 81 Issue: 3 Pages: 551-554
  • DOI: 10.1080/09168451.2016.1249451
  • Peer Reviewed / Open Access
• [Journal Article] The PP-motif in luminal loop 2 of ZnT transporters plays a pivotal role in TNAP activation (2016)
  • Author(s): Fujimoto S, Tsuji T, Fujiwara T, Takeda T, Merriman C, Fukunaka A, Nishito Y, Fu D, Hoch E, Sekler I, Fukue K, Miyamae Y, Masuda S, Nagao M, Kambe T
  • Journal Title: Biochem. J Volume: 473 Issue: 17 Pages: 2611-2621
  • DOI: 10.1042/bcj20160324
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Tetrandrine induces lipid accumulation through blockade of autophagy in a hepatic stellate cell line (2016)
  • Author(s): Miyamae Y, Nishito Y, Nakai N, Nagumo Y, Usui T, Masuda S, Kambe T, Nagao M
  • Journal Title: Biochem Biophys Res Commun Volume: 477 Issue: 1 Pages: 40-46
  • DOI: 10.1016/j.bbrc.2016.06.018
  • Peer Reviewed / Open Access
• [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
• [Presentation] mRNAの成熟阻害活性を指標とした抗ガン活性化合物の探索 (2017)
  • Author(s): 森本 茉莉、北野敦子、鶴岡諒子、増田誠司
  • Organizer: 日本農芸化学会2017年度大会
  • Place of Presentation: 京都女子大学（京都・京都）
  • Year and Date: 2017-03-19
• [Presentation] A Molecular mechanism of selective mRNA export by UAP56 and URH49, the two closely related DEAD-box helicases. (2017)
  • Author(s): Ken-ichi Fujita, Midori Irie, and Seiji Masuda.
  • Organizer: 15th INTERNATIONAL STUDENT SEMINAR
  • Place of Presentation: 芝蘭会館（京都・京都）
  • Year and Date: 2017-02-23
  • Int'l Joint Research
• [Presentation] The Interactome Analysis of PABPN1 in the nucleus of HeLa cell lines (2017)
  • Author(s): Intisar Fouad Ali Mursi, Naoko Fujiwara, Seiji Masuda
  • Organizer: 15th INTERNATIONAL STUDENT SEMINAR
  • Place of Presentation: 芝蘭会館（京都・京都）
  • Year and Date: 2017-02-23
  • Int'l Joint Research
• [Presentation] The Interactome Analysis of PABPN1 in the nucleus of HeLa cell lines 14th international Students’ Seminar (2016)
  • Author(s): Intisar Fouad Ali Mursi, Naoko Fujiwara, Seiji Masuda
  • Organizer: 14th international Students’ Seminar
  • Place of Presentation: Cluj-Napoca, Republic of Croatia
  • Year and Date: 2016-07-26
  • Int'l Joint Research
• [Presentation] A Molecular mechanism to recognize distinct target mRNAs by UAP56 and URH49, the two closely related DEAD-box helicases. (2016)
  • Author(s): Ken-ichi Fujita, and Seiji Masuda
  • Organizer: RNA2016
  • Place of Presentation: 京都国際会議場（京都・京都）
  • Year and Date: 2016-06-28
  • Int'l Joint Research
• [Presentation] 大豆中のmRNA成熟阻害活性の同定・構造活性相関・作用基盤の解明 (2016)
  • Author(s): 増田誠司
  • Organizer: 日本農芸化学会大会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2016-03-28
  • Invited
• [Presentation] A Molecular mechanism to recognize distinct target mRNAs by UAP56 and URH49, the two closely related DEAD-box helicases. (2016)
  • Author(s): Kenichi Fujita, Misa Ito and Seiji Masuda
  • Organizer: The 14th International Student Seminar
  • Place of Presentation: 芝蘭会館（京都府京都市）
  • Year and Date: 2016-03-11
  • Int'l Joint Research
• [Presentation] The interactome analysis of PABPN1 in the nucleus of HeLa cells (2016)
  • Author(s): Intisar F. A. Mulsi, Naoko Fujiwara and Seiji Masuda
  • Organizer: The 14th International Student Seminar
  • Place of Presentation: 芝蘭会館（京都府京都市）
  • Year and Date: 2016-03-11
  • Int'l Joint Research
• [Presentation] mRNA核外輸送因子UAP56とURH49が異なる標的mRNA群を認識する仕組み (2015)
  • Author(s): 藤田賢一、伊藤慶紗、増田誠司
  • Organizer: RNA frontier Meeting
  • Place of Presentation: タカミヤビレッジホテル樹林（山形県山形市）
  • Year and Date: 2015-12-10
• [Presentation] 異なる複合体を形成するmRNA輸送因子UAP56およびURH49の三次元構造解析 (2015)
  • Author(s): 伊藤慶紗、藤田賢一、増田誠司
  • Organizer: 日本分子生物学会・日本生化学会合同大会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-03
• [Presentation] mRNA核外輸送共役因子UAP56ならびにURH49における複合体形成と選択的mRNA輸送の制御基盤 (2015)
  • Author(s): 藤田賢一、伊藤慶紗、増田誠司
  • Organizer: 日本分子生物学会・日本生化学会合同大会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-03
• [Presentation] 哺乳類mRNA核外輸送受容体補因子NXT遺伝子の多様化によるmRNA核外輸送ならびに遺伝子発現の制御 (2015)
  • Author(s): 猪瀬春子、増田誠司
  • Organizer: 日本分子生物学会・日本生化学会合同大会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-03
• [Presentation] mRNA核外輸送因子UAP56ならびにURH49における複合体形成基盤と選択的mRNA輸送制御機構の解明 (2015)
  • Author(s): 藤田賢一、伊藤慶紗、増田誠司
  • Organizer: 日本RNA学会
  • Place of Presentation: ホテルライフォート札幌（北海道札幌市）
  • Year and Date: 2015-07-17
• [Presentation] 異なる複合体を形成するmRNA輸送因子UAP56およびURH49の構造解析 (2015)
  • Author(s): 伊藤慶紗、藤田賢一、増田誠司
  • Organizer: 日本RNA学会
  • Place of Presentation: ホテルライフォート札幌（北海道札幌市）
  • Year and Date: 2015-07-16
• [Presentation] 人工的mRNA核外輸送システムによる組換えタンパク質高効率発現系の構築 (2015)
  • Author(s): 猪瀬春子、増田誠司
  • Organizer: 日本動物細胞工学会
  • Place of Presentation: 東北大学（宮城県仙台市）
  • Year and Date: 2015-07-09
• [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
• [Book] 効率的なmRNA核外輸送系を用いたタンパク質生産法 「動物細胞培養の手法と細胞死・増殖不良・細胞変異を防止する技術」 (2014)
  • Author(s): 岡村真純、平山瑞季、猪瀬春子、増田誠司
  • Total Pages: 584
  • Publisher: 技術情報協会編
• [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/?post_type=labos&p=160
• [Remarks] 京都大学大学院生命科学研究科分子応答機構学増田グループ
  • URL: http://www.bunshioutou.lif.kyoto-u.ac.jp/masuda/index
• [Remarks] 京都大学大学院生命科学研究科分子応答機構学教室
  • URL: http://www.bunshioutou.lif.kyoto-u.ac.jp
• [Remarks] 京都大学大学院生命科学研究科分子応答機構学分野
  • URL: http://www.bunshioutou.lif.kyoto-u.ac.jp