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Development and application of novel chemical modification method for analysis of unusual G-quadruplex structures

Research Project

Project/Area Number 16K17938
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Bio-related chemistry
Research InstitutionUniversity of Miyazaki

Principal Investigator

ISHIZUKA TAKUMI  宮崎大学, 医学部, 助教 (50700085)

Project Period (FY) 2016-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords四重鎖構造 / 核酸 / ヒトテロメアRNA / 化学修飾 / 19F-NMR / フッ素 / グアニン / ヒトテロメアDNA
Outline of Final Research Achievements

In this study, we developed a novel chemical modification method to analyze unusual G-quadruplex structures in living cells. Specifically, in-cell 19F NMR spectroscopy was employed to investigate the structural features of human telomere RNA in Xenopus laevis oocytes. Only one signal was observed in the in-cell 19F NMR spectrum, for which the chemical shift is identical to that observed for the corresponding higher-order G-quadruplex in the in vitro 19F NMR spectrum. We further showed that 19F-NMR can detect DNA/RNA hybrid G-quadruplex structure. We performed a temperature dependent experiment to investigate the stability of the DNA/RNA hybrid G-quadruplex. The result indicated that 19F-NMR can be used to directly observe the melting process of the DNA/RNA hybrid G-quadruplex, which was not easily accessed by traditional methods, such as CD or UV spectroscopy.

Report

(3 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • Research Products

    (24 results)

All 2018 2017 2016 Other

All Journal Article (9 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 9 results,  Open Access: 6 results,  Acknowledgement Compliant: 3 results) Presentation (14 results) (of which Int'l Joint Research: 5 results) Remarks (1 results)

  • [Journal Article] A Simple and Sensitive 19F NMR Approach for Studying the Interaction of RNA G-Quadruplex with Ligand Molecule and Protein2017

    • Author(s)
      H. L. Bao, T. Ishizuka, A. Iwanami, T. Oyoshi, Y. Xu*
    • Journal Title

      ChemistrySelect

      Volume: - Issue: 15 Pages: 4170-4175

    • DOI

      10.1002/slct.201700711

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structure-Dependent Binding of hnRNPA1 to Telomere RNA2017

    • Author(s)
      Liu Xiao、Ishizuka Takumi、Bao Hong-Liang、Wada Kei、Takeda Yuma、Iida Keisuke、Nagasawa Kazuo、Yang Danzhou、Xu Yan
    • Journal Title

      Journal of the American Chemical Society

      Volume: 139 Issue: 22 Pages: 7533-7539

    • DOI

      10.1021/jacs.7b01599

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Antiparallel RNA G-quadruplex Formed by Human Telomere RNA Containing 8-Bromoguanosine2017

    • Author(s)
      Chao-Da Xiao, Takumi Ishizuka & Yan Xu
    • Journal Title

      Scientific Reports

      Volume: 7 Issue: 1 Pages: 6695-6695

    • DOI

      10.1038/s41598-017-07050-w

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A multi-functional guanine derivative for studying the DNA G-quadruplex structure2017

    • Author(s)
      Takumi Ishizuka, Pei-Yan Zhao, Hong-Liang Bao and Yan Xu
    • Journal Title

      Analyst

      Volume: 142 Issue: 21 Pages: 4083-4088

    • DOI

      10.1039/c7an00941k

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Studying DNA G-Quadruplex Aptamer by 19F NMR2017

    • Author(s)
      Takumi Ishizuka, Atsushi Yamashita, Yujiro Asada, and Yan Xu
    • Journal Title

      ACS Omega

      Volume: 2 Issue: 12 Pages: 8843-8848

    • DOI

      10.1021/acsomega.7b01405

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Characterization of Human Telomere RNA G-Quadruplex Structures in vitro and in Living Cells using 19F NMR Spectroscopy2017

    • Author(s)
      H. L. Bao, T. Ishizuka, T. Sakamoto, K. Fujimoto, T. Uechi, N. Kenmochi, Y. Xu*
    • Journal Title

      Nucleic Acids Res.

      Volume: - Issue: 9 Pages: 5501-5511

    • DOI

      10.1093/nar/gkx109

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Unusual Topological RNA Architecture with an Eight-Stranded Helical Fragment Containing A-, G-, and U-Tetrads2017

    • Author(s)
      C. D. Xiao, T. Ishizuka, X. Q. Zhu, Y. Li, H. Sugiyama, Y. Xu*
    • Journal Title

      J. Am. Chem. Soc.

      Volume: 139 Issue: 7 Pages: 2565-2568

    • DOI

      10.1021/jacs.6b12274

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Fluorescence Imaging of Chromosomal DNA using Click Chemistry2016

    • Author(s)
      T. Ishizuka, H. S. Liu, K. Ito, Y. Xu*
    • Journal Title

      Sci. Rep.

      Volume: 6 Issue: 1 Pages: 33217-33217

    • DOI

      10.1038/srep33217

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Click Chemistry Takes a Snapshot of DNA-RNA Hybrid G-Quadruplex in Living Cells2016

    • Author(s)
      Takumi Ishizuka, Yan Xu
    • Journal Title

      Journal of Nature and Science

      Volume: 2

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Chemical probing of G-quadruplex structures by modified nucleosides2018

    • Author(s)
      Takumi Ishizuka, Yan Xu
    • Organizer
      日本化学会第98春季年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] RNA四重鎖構造を解析する化学プローブの開発2018

    • Author(s)
      大立目 真臣, 石塚 匠, 徐 岩
    • Organizer
      日本化学会第98春季年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] hnRNPA1のヒトテロメアRNAへの構造特異的な結合2018

    • Author(s)
      竹田 悠馬, 劉 暁, 石塚 匠, 徐 岩
    • Organizer
      日本化学会第98春季年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Studying G-quadruplex structures by 19F NMR2018

    • Author(s)
      Hong-Liang Bao, Takumi Ishizuka, Yan Xu
    • Organizer
      日本化学会第98春季年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Investigation of the thermodynamic stability and structure of oligonucleotides containing 2’-O-methyl-8-methylguanosine by CD and NMR2018

    • Author(s)
      Thananjeyan Balasubramaniyam, Takumi Ishizuka, Yan Xu
    • Organizer
      日本化学会第98春季年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Development of New Chemical Modification Method for Analysis of Unique G-Quadruplex Structures2017

    • Author(s)
      Takumi Ishizuka, Hong-Liang Bao, Yan Xu
    • Organizer
      日本化学会第97春季年会
    • Place of Presentation
      慶應義塾大学
    • Year and Date
      2017-03-16
    • Related Report
      2016 Research-status Report
  • [Presentation] Chemical Probe for Studying Human Telomere RNA G-quadruplex2017

    • Author(s)
      Takumi Ishizuka, Masaomi Otatsume, Yan Xu
    • Organizer
      The 44th International Symposium on Nucleic Acid Chemistry
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Novel RNA Architecture Formed by Eight RNA Strands Containing Fourteen G- A- and U-tetrads2017

    • Author(s)
      Chao-Da Xiao, Takumi Ishizuka, Yan Xu
    • Organizer
      The 44th International Symposium on Nucleic Acid Chemistry
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] To investigate human telomere DNA G-quadruplexes structures by 19F NMR2017

    • Author(s)
      Hong-Liang Bao, Takumi Ishizuka, Yan Xu
    • Organizer
      The 44th International Symposium on Nucleic Acid Chemistry
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] RNA四重鎖構造を解析するケミカルプローブの開発2017

    • Author(s)
      石塚 匠, 大立目 真臣, 徐 岩
    • Organizer
      第11回バイオ関連化学シンポジウム
    • Related Report
      2017 Annual Research Report
  • [Presentation] A multi-functional guanine derivative for studying DNA G-quadruplex structure in vitro and in living cells2017

    • Author(s)
      Takumi Ishizuka, Yan Xu
    • Organizer
      G4thering-6th International Meeting on Quadruplex Nucleic Acids
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Conformationally Restricted Guanosine Analogues Induce Topological Change in Human Telomeric DNA G-Quadruplexes2016

    • Author(s)
      Takumi Ishizuka, Yan Xu
    • Organizer
      The 43th International Symposium on Nucleic Acid Chemistry
    • Place of Presentation
      熊本大学
    • Year and Date
      2016-09-27
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] 修飾グアノシン誘導体により形成する核酸高次構造2016

    • Author(s)
      石塚 匠, 徐 岩
    • Organizer
      第10回バイオ関連化学シンポジウム
    • Place of Presentation
      石川県立音楽堂
    • Year and Date
      2016-09-07
    • Related Report
      2016 Research-status Report
  • [Presentation] 光架橋反応を用いたヒトテロメアRNA四重鎖とhnRNPA1の相互作用解析2016

    • Author(s)
      石塚 匠, 徐 岩
    • Organizer
      第38回日本光医学・光生物学会
    • Place of Presentation
      京都リサーチパーク
    • Year and Date
      2016-07-22
    • Related Report
      2016 Research-status Report
  • [Remarks] 宮崎大学医学部機能制御学講座物質科学分野ホームページ

    • URL

      http://www.med.miyazaki-u.ac.jp/MMCCHEM/

    • Related Report
      2017 Annual Research Report

URL: 

Published: 2016-04-21   Modified: 2019-03-29  

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