Mechanistic Studies on the Photooxidation of Methylated Cytosine in DNA and Its Application to Novel DNA Methylation Analysis

• Project/Area Number: 21710228
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Living organism molecular science
• Research Institution: Kyoto University
• Principal Investigator: YAMADA Hisatsugu Kyoto University, 先端医工学研究ユニット, 助教 (80512764)
• Project Period (FY): 2009 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2009: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 機能性核酸 / 光増感反応 / DNA内電荷移動反応 / 5-メチルシトシン / 同位体効果 / DNA内電荷移動 / DNAメチル化 / 修飾シトシン塩基 / 重水素同位体効果

Research Abstract

Our interest focuses on the potential application of remarkable photooxidative reactivity of 5-methylcytosine (^mC) to methylation-mapping system and on further understanding the mechanism of photooxidation at ^mC in DNA. In the project, we revealed that (1) incorporation of an anthraquinone (AQ) sensitizer into a DNA strand enhanced one-electron photooxidation, leading to efficient strand cleavage at the ^mC site, (2) deprotonation at the C5-methyl group of an intermediate ^mC radical cation may occur as a key elementary reaction in the AQ-photooxidative strand cleavage at the ^mC, and (3) DNA strand cleavage at multiple ^mC residues was induced by AQ-photoinjected positive charge (hole) transfer through DNA duplex. The results obtained may provide a guide for the molecular design of highly sensitive and selective photochemical methods for the identification and analysis of ^mC in DNA.

Research Products

• [Journal Article] Preparation of Functionalized Oligodeoxynucleotides and Photochemical One-Electron Oxidation of 5-Methylcytosine in DNA (2011)
  • Author(s): Tanabe, K., Yamada, H., Nishimoto S.
  • Journal Title: J.Synth.Org.Chem. in press
  • NAID: 10029438334
  • Peer Reviewed
• [Journal Article] Anthraquinone-Sensitized Photooxidation of 5-Methylcytosine in DNA Leading to Piperidine-Induced Efficient StrandCleavage (2011)
  • Author(s): Yamada, H., Kitauchi, Y., Tanabe, K., Ito, T., Nishimoto, S.
  • Journal Title: Chem.Eur.J. 17 Pages: 2225-2235
  • Peer Reviewed
• [Journal Article] Photoinduced Electron Injection into DNA by N-Cyclopropyl-1-aminonaphthalene (2011)
  • Author(s): Ito, T., Uchida, T., Tanabe, K., Nishimoto, S.
  • Journal Title: J.Photochem.Photobiol.A Chem. 219 Pages: 115-121
  • Peer Reviewed
• [Journal Article] Anthraquinone-Sensitized Photooxidation of 5-Methylcytosine in DNA Leading to Piperidine-Induced Efficient Strand Cleavage (2011)
  • Author(s): Yamada, H.; Kitauchi, Y.; Tanabe, K.; Ito, T.; Nishimoto, S.
  • Journal Title: Chem.Eur.J. Volume: 17 Pages: 2225-2235
  • Peer Reviewed
• [Journal Article] Photoinduced Electron Injection into DNA by N-Cyclopropyl-1-aminonaphthalene (2011)
  • Author(s): Ito, T.; Uchida, T.; Tanabe, K.; Yamada, H.; Nishimoto, S.
  • Journal Title: J.Photochem.Photobiol.A Chem. Volume: 219 Pages: 115-121
  • Peer Reviewed
• [Journal Article] Photo-Induced Electron Transfer Reaction in Diaminostilbene-Tethered DNA Duplexes (2009)
  • Author(s): Ito, T., Hayashi, A., Uchida, T., Tanabe, K., Yamada, H., Nishimoto, S.
  • Journal Title: Nucleic Acids Symp.Ser. 53 Pages: 201-202
• [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] 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] Preparation of Functionalized Oligodeoxynucleotides and Photochemical One-Electron Oxidation of 5-Methylcytosine in DNA
  • Author(s): Tanabe, K.; Yamada, H.; Nishimoto S.
  • Journal Title: J.Synth.Org.Chem. Volume: (In press)
  • NAID: 10029438334
  • Peer Reviewed
• [Presentation] Efficient one-electron photooxidation and site-selective strand cleavage at 5-methylcytosine in DNA by oligonucleotide bearing 9,10-anthraquinone sensitizer (2010)
  • Author(s): Kitauchi, Y., Yamada, H., Tanabe, K., Ito, T., Nishimoto, S.
  • Organizer: Pacifichem 2010
  • Place of Presentation: Honolulu, USA
  • Year and Date: 2010-12-19
• [Presentation] Selective trapping at cytosine derivatives of photosensitizer-injected and migrated hole in DNA (2010)
  • Author(s): Kurata, M., Yamada, H., Tanabe, K., Ito, T., Nishimoto, S.
  • Organizer: Pacifichem 2010
  • Place of Presentation: Honolulu, USA
  • Year and Date: 2010-12-19
• [Presentation] Efficient one-electron photooxidation and site-selective strand cleavage at 5-methylcytosine in DNA by oligonucleotide bearing 9, 10-anthraquinone sensitizer (2010)
  • Author(s): Kitauchi, Y.; Yamada, H.; Tanabe, K.; Ito, T.; Nishimoto, S.
  • Organizer: Pacifichem 2010
  • Place of Presentation: Honolulu, USA
  • Year and Date: 2010-12-19
• [Presentation] Selective trapping at cytosine derivatives of photosensitizerinjected and migrated hole in DNA (2010)
  • Author(s): Kurata, M.; Yamada, H.; Tanabe, K.; Ito, T.; Nishimoto, S.
  • Organizer: Pacifichem 2010
  • Place of Presentation: Honolulu, USA
  • Year and Date: 2010-12-19
• [Presentation] Efficient Hole Trapping at C(5)-Substituted Cytosine in the Photosensitizer-Injected Positive Charge Transfer through DNA Duplex (2010)
  • Author(s): Kurata, M., Yamada, H., Tanabe, K., Ito, T., Nishimoto, S.
  • Organizer: 7thInternational Symposium on Nucleic Acids Chemistry
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2010-11-11
• [Presentation] One-Electron Oxidation at 5-Methylcytosine Induced by thePhotosensitizer-Injected Positive ChargeTransfer through DNA (2010)
  • Author(s): Kitauch, Y., Tanabe, K., Ito, T., Nishimoto, S.
  • Organizer: 7th International Symposium on Nucleic Acids Chemistry
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2010-11-11
• [Presentation] Efficient Hole Trapping at C(5)-Substituted Cytosine in the Photosensitizer-Injected Positive Charge Transfer through DNA Duplex (2010)
  • Author(s): Kurata, M.; Yamada, H.; Tanabe, K.; Ito, T.; Nishimoto, S.
  • Organizer: 7th International Symposium on Nucleic Acids Chemistry
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2010-11-11
• [Presentation] One-Electron Oxidation at 5-Methylcytosine Induced by the Photosensitizer-Injected Positive Charge Transfer through DNA (2010)
  • Author(s): Kitauchi, Y.; Yamada.H.; Tanabe, K.; Ito, T.; Nishimoto, S.
  • Organizer: 7th International Symposium on Nucleic Acids Chemistry
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 2010-11-11
• [Presentation] メチル化DNAの光増感酸化反応に及ぼす重水素同位体効果 (2010)
  • Author(s): 山田久嗣、北内佑哉、田邉一仁、伊藤健雄、西本清一
  • Organizer: 第90回日本化学会春季年会
  • Place of Presentation: 東大阪
  • Year and Date: 2010-03-27
• [Presentation] メチル化DNAの光増感酸化反応に及ぼす重水素同位体効果 (2010)
  • Author(s): 山田久嗣、北内佑哉、田邉一仁、伊藤健雄、西本清一
  • Organizer: 第90回日本化学会春季年会
  • Place of Presentation: 東大阪市
  • Year and Date: 2010-03-27
• [Presentation] 光化学反応を利用したDNAメチル化検出法の開発 (2010)
  • Author(s): 山田久嗣
  • Organizer: バイオ計測・試薬研究会「バイオ計測・試薬分野における若手科学者・技術者のシーズ・構想発表会」
  • Place of Presentation: 京都
  • Year and Date: 2010-03-10
• [Presentation] Efficient Photooxidative Strand Cleavage at 5-Methylcytosine in DNA by Sensitization with 9,10-Anthraquinone-Tethered Oligonucleotides (2009)
  • Author(s): 山田久嗣、北内佑哉、田邉一仁、伊藤健雄、西本清一
  • Organizer: 6th International Symposium on Nucleic Acids Chemistry
  • Place of Presentation: Takayama, Japan
  • Year and Date: 2009-09-29
• [Presentation] アントラキノン光増感剤を導入したオリゴDNA : 5-メチルシトシン塩基部位での選択的DNA切断 (2009)
  • Author(s): 北内佑哉、山田久嗣、田邉一仁、伊藤健雄、西本清一
  • Organizer: 第24回生体機能関連化学シンポジウム
  • Place of Presentation: 福岡市
  • Year and Date: 2009-09-14
• [Presentation] C(5)位に置換基を導入した修飾シトシン含有DNAオリゴマーの合成と四重鎖形成特性 (2009)
  • Author(s): 藏田真之、山田久嗣、田邉一仁、伊藤健雄、山田久嗣、西本清一
  • Organizer: 第24回生体機能関連化学シンポジウム若手フォーラム
  • Place of Presentation: 福岡
• [Presentation] アントラキノン光増感剤を導入したオリゴDNA:5-メチルシトシン塩基部位での選択的DNA切断 (2009)
  • Author(s): 北内佑哉、田邉一仁、伊藤健雄、西本清一
  • Organizer: 第24回生体機能関連化学シンポジウム
  • Place of Presentation: 福岡
• [Presentation] Efficient Photooxidative Strand Cleavage at 5-Methylcytosine in DNA by Sensitization with9,10-Anthraquinone-Tethered Oligonucleotides (2009)
  • Author(s): Yamada, H., Kitauchi, Y., Tanabe, K., Ito, T., Nishimoto, S.
  • Organizer: 6th Symposium on Nucleic Acids Chemistry
  • Place of Presentation: Takayama, Japan
• [Presentation] 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.
  • Organizer: 6th International Symposium on Nucleic Acids Chemistry
  • Place of Presentation: Takayama, Japan
• [Presentation] Photochemical Approach to Discrimination of Methylated Cytosine in DNA : pH Effect on theNaphthoquinone-Sensitized One-Electron Photooxidation of 5-Methylcytosine (2009)
  • Author(s): Yamada, H., Kitauchi, Y., Tanabe, K., Ito, T., Nishimoto, S.
  • Organizer: XXIV International Conference on Photochemistry
  • Place of Presentation: Toledo, Spain