Detection of 5-methylcytosine in DNA by functionalized oligodeoxynucleotides

• Project/Area Number: 20750134
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Chemistry related to living body
• Research Institution: Kyoto University
• Principal Investigator: TANABE Kazuhito Kyoto University, 工学研究科, 准教授 (40346086)
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2009: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2008: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: 生物有機化学 / メチル化シトシン / DNA / 光反応 / 脱プロトン化 / 鎖切断 / ナフトキノン / 光酸化反応 / DNA切断

Research Abstract

Although the NQ-photosensitized oxidation accompanied by selective strand cleavage at ^mC in DNA is an attractive method for identification of the methylation site in DNA, the relatively lower sensitivity of the detection method due to the less efficiency in the photooxidation of ^mC still remains as a major subject to be improved for establishing a status of general availability. In this study, we characterized reaction mechanism of the NQ-photosensitized one-electron oxidation of 5-methyl-2-deoxycytidine (d^mC) in order to obtain insight into the design of photochemical system involving the ^mC-selective strand cleavage with high sensitivity. We confirmed that deprotonation of d^mC radical cation and formation of methyl radical species are key reaction for the present photoreaction.

Research Products

• [Journal Article] Efficient photooxidative strand cleavage at 5-methylcytosine in DNA by sensitization with 9, 10-anthraquinone-tethered oligonucleotides (2009)
  • Author(s): Yamada, H.; Kitauchi, Y.; Tanabe, K.; Ito, T.; Nishimoto, S.
  • Journal Title: Nucleic Acids Symp. Ser. 53 Pages: 209-210
• [Journal Article] Photoelectrochemical identification of 5-methylcytosine modification in DNA: combination of photosensitization and enzymatic cleavage (2009)
  • Author(s): Tanabe, K.; Yamada, H.; Ito, T.; Nishimoto, S.
  • Journal Title: Nucleic Acids Symp. Ser. 53 Pages: 205-206
• [Journal Article] Monitoring of biological one-electron reduction by ^<19>F-NMR using hypoxia selective activation of a ^<19>F-labeled indolequinone derivative (2009)
  • Author(s): Tanabe, K.; Harada, H.; Narazaki, M.; Tanaka, K.; Inafuku, K.; Komatsu, H.; Ito, T.; Yamada, H.; Chujo, Y.; Matsuda, T.; Hiraoka, M.; Nishimoto, S.
  • Journal Title: J. Am. Chem. Soc. 131 Pages: 15982-15983
  • Peer Reviewed
• [Journal Article] Preparation and Fluorescence Properties of Fluorophore-labeled Avidin-biotin System Immobilized on Fe_3O_4 Nanoparticles through Functional Indolequinone Linker (2009)
  • Author(s): Hirata, N.; Tanabe, K.; Narita, A.; Tanaka, K.; Naka, K.; Chujo, Y.; Nishimoto, S.
  • Journal Title: Bioorg. Med. Chem. 17 Pages: 3775-3781
  • Peer Reviewed
• [Journal Article] Radiolytic activation of cytarabine prodrug possessing 2-oxoalkyl group: one-electron reduction and cytotoxicity characteristics (2009)
  • Author(s): Hirata, N.; Fujisawa, Y.; Tanabe, K.; Harada, H.; Hiraoka, M.; Nishimoto, S.
  • Journal Title: Org. Biomol. Chem. 7 Pages: 651-654
  • NAID: 120001624118
  • Peer Reviewed
• [Journal Article] Radiolytic activation of cytarabine prodrug possessing 2-oxoalkyl group : one-electron reduction and (2009)
  • Author(s): Hirata, N.; Fujisawa, Y.; Tanabe, K.; Harada, H.; Hiraoka, M.;
  • Journal Title: Org.Biomol.Chem 7 Pages: 651-654
  • Peer Reviewed
• [Journal Article] Preparation and Fluorescence Properties of Fluorophore-labeled Avidin-biotin System Immobilized on Fe3O4 (2009)
  • Author(s): Hirata, N.; Tanabe, K.; Narita, A.; Tanaka, K.; Naka, K.; Chujo
  • Journal Title: Bioorg.Med.Chem. 17 Pages: 3775-3781
  • Peer Reviewed
• [Journal Article] Monitoring of biological one-electron reduction by 19F-NMR using hypoxia selective activation of a 19F-labeled (2009)
  • Author(s): Tanabe, K.; Harada, H.; Narazaki, M.; Tanaka, K.; Inafuku, K.;
  • Journal Title: J.Am.Chem.Soc. 131 Pages: 15982-15983
  • Peer Reviewed
• [Journal Article] Radiolytic one-electron reduction characteristics of tyrosine derivative caged by 2-oxopropyl group (2008)
  • Author(s): Tanabe, K.; Ebihara, M.; Hirata, N.; Nishimoto, S.
  • Journal Title: Bioorg. Med. Chem. Lett. 18 Pages: 6126-6129
  • NAID: 120001624120
  • Peer Reviewed
• [Journal Article] Radiation- and photo-induced activation of 5-fluorouracil prodrugs as a strategy for the selective treatment of solid tumors (2008)
  • Author(s): Ito, T.; Tanabe, K.; Yamada, H.; Hatta, H.; Nishimoto, S.
  • Journal Title: molecules 13 Pages: 2370-2384
  • NAID: 120001462592
  • Peer Reviewed
• [Journal Article] The pH effect on the naphthoquinone-photosensitized oxidation of 5-methylcytosine (2008)
  • Author(s): Yamada, H.; Tanabe, K.; Ito, T.; Nishimoto, S.
  • Journal Title: Chem. Eur. J. 14 Pages: 10453-10461
  • Peer Reviewed
• [Journal Article] Photoelectrochemical evaluation of pH effect on hole transport through triplex-forming DNA immobilized on a gold electrode (2008)
  • Author(s): Haruna, K.; Iida, H.; Tanabe, K.; Nishimoto, S.
  • Journal Title: Org. Biomol. Chem. 6 Pages: 1613-1617
  • NAID: 120001624119
  • Peer Reviewed
• [Journal Article] One-Electron reductive template-directed ligation of oligodeoxynucleotides possessing disulfide bond (2008)
  • Author(s): Tanabe, K.; Kuraseko, E.; Yamamoto, Y.; Nishimoto, S.
  • Journal Title: J. Am. Chem. Soc. 130 Pages: 6302-6303
  • Peer Reviewed
• [Journal Article] Fluorometric identification of 5-methylcytosine modification in DNA: combination of photosensitized oxidation and invasive cleavage (2008)
  • Author(s): Yamada, H.; Tanabe, K.; Nishimoto, S.
  • Journal Title: Bioconjugate Chem. 19 Pages: 20-23
  • Peer Reviewed
• [Journal Article] The pH effect on the naphthoquinone-photosensitized oxidation of 5-methylcytosine (2008)
  • Author(s): Yamada, H.; Tanabe, K. Ito, T.; Nishimoto, S.
  • Journal Title: Chem. Eur. J. 14 Pages: 10453-10461
  • Peer Reviewed
• [Journal Article] Radiation-and photo-induced activation of 5-fluorouracil prodrugs as strategy for the selective (2008)
  • Author(s): Ito, T.; Tanabe, K.; Yamada, H. Hatta, H.; Nishimoto, S.
  • Journal Title: molecules 13 Pages: 2370-2384
  • Peer Reviewed
• [Journal Article] One-Electron reductive template-directed ligation of oligodeoxynucleotides possessing disulfide bond (2008)
  • Author(s): Tanabe, K.; Kuraseko, Yamamoto, Y.; Nishimoto, S.
  • Journal Title: J. Am. Chem. Soc. 130 Pages: 6302-6303
  • Peer Reviewed
• [Presentation] Photoelectrochemical identification of 5-methylcytosine modification in DNA : combination of photosensitization (2009)
  • Author(s): Tanabe, K.; Yamada, H.; Ito, T.; Nishimoto, S.
  • Organizer: 第6回国際核酸化学シンポジウム
  • Place of Presentation: 高山市
  • Year and Date: 2009-09-29
• [Presentation] pH effect on the one-electron photooxidation of 5-methylcytosine with naphthoquinone sensitizer (2008)
  • Author(s): Yamada, H.; Tanabe, K.; Ito, T.; Nishimoto, S.
  • Organizer: 第35回核酸化学シンポジウム
  • Place of Presentation: 京都
  • Year and Date: 2008-09-11
• [Presentation] pH effect on the one-electron photooxidation of 5-methylcytosine with naphthoquinone sensitizer (2008)
  • Author(s): Yamada, H.; Tanabe, K.; Ito, T.; Nishimoto, S.
  • Organizer: 35th International Symposium on Nucleic Acids Chemistry
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2008-09-11
• [Presentation] DNA中メチル化シトシンの電気化学的検出 (2008)
  • Author(s): 田邉一仁・山田久嗣・西本清一
  • Organizer: 第30回光医学・光生物学会
  • Place of Presentation: 松江
  • Year and Date: 2008-07-11
• [Remarks] ホームページでの研究紹介
  • URL: http://www.ehcc.kyoto-u.ac.jp/eh32/home/web-content/index-j.htm