Structural probing of RNA G-quadruplex structures in vivo

• Project/Area Number: 21K05314
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 37030:Chemical biology-related
• Research Institution: University of Miyazaki
• Principal Investigator: Ishizuka Takumi 宮崎大学, 医学部, 学部講師 (50700085)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2023: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: グアニン四重鎖 / RNA / セリウム / グアニン四重鎖構造

Outline of Research at the Start

本研究ではRNAグアニン四重鎖構造(rG4)をヌクレオチドレベルの分解能で検出可能な技術を開発し、細胞内rG4構造の解析を行う。また、異なる細胞種や異なる細胞周期において本技術を応用し、細胞内のrG4構造の時空間的情報を取得する。その結果に基づきrG4に関する新たな機能を明らかにする。

Outline of Final Research Achievements

RNA G-quadruplexes (rG4) have important biological roles and are expected as an attractive therapeutic target for drug design. The structural analysis of rG4 provides significant information for a better understanding of the function of rG4. We found that Ce(IV)/EDTA complex as a molecular scissor that can be effectively and selectively cleaved specific position of nucleotides in rG4. We demonstrated that the loop regions of rG4 are specifically cleaved by the Ce(IV)/EDTA complex. This strategy provides a useful method to probe rG4 at nucleotide resolution. The current study is the first step in providing insight into the structure-dependent Ce(IV)/EDTA reactivity in rG4. The method will likely make possible facile nucleotide resolution analysis of RNA structures and provides a concept for targeting rG4 related to disease.

Academic Significance and Societal Importance of the Research Achievements

rG4は神経疾患に関与しており、今回開発したrG4解析法は、形成メカニズムをはじめとした未解明の機能の理解が進むものと期待される。Ce(IV)/EDTA錯体の生体親和性を考慮するとin vivoでのrG4の解析が可能であり、本手法を利用した診断法への応用も期待できる。

Research Products

• [Journal Article] Protective role of estrogen through G-protein coupled receptor 30 in a colitis mouse model (2024)
  • Author(s): Fidya, Narantsog Choijookhuu, Makoto Ikenoue, Koichi Yano, Yu Yamaguma, Shinichiro Shirouzu, Kengo Kai, Takumi Ishizuka, Yoshitaka Hishikawa
  • Journal Title: Histochemistry and Cell Biology Volume: 161 Issue: 1 Pages: 81-93
  • DOI: 10.1007/s00418-023-02235-z
  • Peer Reviewed
• [Journal Article] The crucial role of SETDB1 in structural and functional transformation of epithelial cells during regeneration after intestinal ischemia reperfusion injury (2024)
  • Author(s): Makoto Ikenoue, Narantsog Choijookhuu, Koichi Yano, Fidya, Nobuyasu Takahashi, Takumi Ishizuka, Shinichiro Shirouzu, Yu Yamaguma, Kengo Kai, Kazuhiro Higuchi, Akira Sawaguchi, Atsushi Nanashima, Yoshitaka Hishikawa
  • Journal Title: Histochemistry and Cell Biology Volume: 161 Issue: 4 Pages: 325-336
  • DOI: 10.1007/s00418-023-02263-9
  • Peer Reviewed
• [Journal Article] HMGB2 Promotes De Novo Lipogenesis to Accelerate Hepatocyte Proliferation During Liver Regeneration (2024)
  • Author(s): Narantsog Choijookhuu, Koichi Yano, Baljinnyam Lkham-Erdene, Shinichiro Shirouzu, Toshiki Kubota, Fidya, Takumi Ishizuka, Kengo Kai, Etsuo Chosa, Yoshitaka Hishikawa
  • Journal Title: Journal of Histochemistry & Cytochemistry Volume: - Issue: 4 Pages: 245-264
  • DOI: 10.1369/00221554241241569
  • Peer Reviewed / Open Access
• [Journal Article] A Highly Sensitive Detection System for <i>in situ</i> Hybridization Using Fluorescence Resonance Energy Transfer (FRET) Phenomenon (2023)
  • Author(s): 石塚 匠, Narantsog Choijookhuu, 柴田 恭明, 小路 武彦, 菱川 善隆
  • Journal Title: KENBIKYO Volume: 58 Issue: 3 Pages: 117-122
  • DOI: 10.11410/kenbikyo.58.3_117
  • ISSN: 1349-0958, 2434-2386
  • Year and Date: 2023-12-30
  • Peer Reviewed
• [Journal Article] An Advanced Detection System for <i>In Situ</i> Hybridization Using a Fluorescence Resonance Energy Transfer-based Molecular Beacon Probe (2022)
  • Author(s): Narantsog Choijookhuu, Yasuaki Shibata, Takumi Ishizuka, Yan Xu, Takehiko Koji, Yoshitaka Hishikawa
  • Journal Title: ACTA HISTOCHEMICA ET CYTOCHEMICA Volume: 55 Issue: 5 Pages: 119-128
  • DOI: 10.1267/ahc.22-00075
  • ISSN: 0044-5991, 1347-5800
  • Year and Date: 2022-10-28
  • Peer Reviewed / Open Access
• [Journal Article] Click chemistry for fluorescence imaging via combination of a BODIPY-based ‘turn-on’ probe and a norbornene glucosamine (2022)
  • Author(s): Zhiyong He, Takumi Ishizuka, Yoshitaka Hishikawa, Yan Xu
  • Journal Title: Chemical Communications Volume: 58 Issue: 89 Pages: 12479-12482
  • DOI: 10.1039/d2cc05359d
  • Peer Reviewed / Open Access
• [Journal Article] Pivotal role of High-Mobility Group Box 2 in ovarian folliculogenesis and fertility (2022)
  • Author(s): Shinichiro Shirouzu, Naohiro Sugita, Narantsog Choijookhuu, Yu Yamaguma, Kanako Takeguchi, Takumi Ishizuka, Mio Tanaka, Fidya, Kengo Kai, Etsuo Chosa, Yoshihiro Yamashita, Chihiro Koshimoto, Yoshitaka Hishikawa
  • Journal Title: Journal of Ovarian Research Volume: 15 Issue: 1 Pages: 133-133
  • DOI: 10.1186/s13048-022-01071-4
  • Peer Reviewed / Open Access
• [Journal Article] Crucial role of high-mobility group box 2 in mouse ovarian follicular development through estrogen receptor beta (2022)
  • Author(s): Yu Yamaguma, Naohiro Sugita, Narantsog Choijookhuu, Koichi Yano, Deokcheol Lee, Makoto Ikenoue, Fidya, Shinichiro Shirouzu, Takumi Ishizuka, Mio Tanaka, Yoshihiro Yamashita, Etsuo Chosa, Noboru Taniguchi, Yoshitaka Hishikawa
  • Journal Title: Histochemistry and Cell Biology Volume: 157 Issue: 3 Pages: 359-369
  • DOI: 10.1007/s00418-022-02074-4
  • Peer Reviewed / Open Access
• [Presentation] 肝再生過程におけるDNA四重鎖構造の肝細胞周期への関与 (2024)
  • Author(s): 石塚 匠, Narantsog Choijookhuu, 白水 慎一郎, Fidya, 甲斐 健吾, 菱川 善隆
  • Organizer: 第129回 日本解剖学会総会・全国学術集会
• [Presentation] マウス肝再生過程におけるグアニン四重鎖構造の発現動態研究 (2023)
  • Author(s): 石塚 匠, Narantsog Choijookhuu, 白水 慎一郎, Fidya, 甲斐 健吾, 菱川 善隆
  • Organizer: 第64回日本組織細胞化学会総会・学術集会
• [Presentation] 肝再生過程におけるグアニン四重鎖構造の肝細胞増殖への関与 (2023)
  • Author(s): 石塚 匠, Narantsog Choijookhuu, 白水 慎一郎, Fidya, 甲斐 健吾, 菱川 善隆
  • Organizer: 日本解剖学会 第79回 九州支部学術集会
• [Presentation] マウス肝再生過程におけるDNA四重鎖構造の肝細胞周期への関与 (2023)
  • Author(s): 石塚 匠, Narantsog Choijookhuu, 白水 慎一郎, Fidya, 甲斐 健吾, 菱川 善隆
  • Organizer: 第65回日本顕微鏡学会九州支部総会・学術講演会
• [Presentation] 肝再生モデルマウスを用いたグアニン四重鎖構造の機能解明 (2022)
  • Author(s): 石塚 匠, Narantsog Choijookhuu, 矢野 公一, 池ノ上 実, 山隈 優, 白水 慎一郎, Fidya, 田中 美緒, 菱川 善隆
  • Organizer: 第39回分子病理学研究会
• [Presentation] マウス卵巣におけるHMGB2の卵胞形成への関与 (2022)
  • Author(s): 山隈 優, Narantsog Choijookhuu, 矢野 公一, 池ノ上 実, 甲斐 健吾, Fidya, 白水 慎一郎, 田中 美緒, 石塚 匠, 菱川 善隆
  • Organizer: 第39回分子病理学研究会
• [Presentation] 蛍光共鳴エネルギー移動（FRET)を利用したin situ hybridization法 (2022)
  • Author(s): 石塚 匠, Narantsog Choijookhuu, 柴田 恭明, 徐 岩, 小路 武彦, 菱川 善隆
  • Organizer: 第64回日本顕微鏡学会九州支部集会
• [Presentation] Estrogen signaling inhibits intestinal epithelial remodeling through G-Protein Coupled Receptor 30 in DSS-induced colitis in mice (2022)
  • Author(s): Fidya, Narantsog Choijookhuu, Makoto Ikenoue, Koichi Yano, Yu Yamaguma, Shinichiro Shirouzu, Kengo Kai, Takumi Ishizuka, Yoshitaka Hishikawa
  • Organizer: 第128回日本解剖学会総会・全国学術集会
• [Presentation] マウス卵巣における HMGB2 の卵胞形成への関与 (2022)
  • Author(s): 山隈 優, Narantsog Choijookhuu, 甲斐 健吾, Fidya, 白水 慎一郎, 石塚 匠, 菱川 善隆
  • Organizer: 第128回日本解剖学会総会・全国学術集会
• [Presentation] 蛍光共鳴エネルギー移動 （FRET)を利用したin situ hybridizationの実際 (2021)
  • Author(s): 菱川 善隆, Narantsog Choijookhuu, 石塚 匠, 柴田 恭明, 小路 武彦
  • Organizer: 第62回日本組織細胞化学会総会・学術集会
• [Book] 組織細胞化学：In situ hybridization法 (2023)
  • Author(s): 菱川 善隆, Narantsog Choijookhuu, 石塚 匠, 柴田 恭明, 小路 武彦
  • Total Pages: 11
  • Publisher: 中西印刷
• [Book] In situ hybridization法 (2022)
  • Author(s): 菱川 善隆, Narantsog Choijookhuu, 石塚 匠, 柴田 恭明, 小路 武彦
  • Total Pages: 258
  • Publisher: 中西印刷
• [Book] 病理と臨床 (2022)
  • Author(s): 菱川 善隆, Narantsog Choijookhuu, 石塚 匠, 柴田 恭明, 小路 武彦
  • Total Pages: 424
  • Publisher: 文光堂
• [Remarks] 宮崎大学研究者データベース
  • URL: https://srhumdb.miyazaki-u.ac.jp/html/100001556_ja.html
• [Remarks] Researchmap
  • URL: https://researchmap.jp/takumiishizuka