Analyses of oxidative stress-dependent formation of MUTYH/MutSalpha complex

• Project/Area Number: 26340025
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Kyushu University
• Principal Investigator: NAKATSU Yoshimichi 九州大学, 医学研究院, 准教授 (00207820)
• Research Collaborator: HAYASHIDA Genki 九州大学, システム生命学府 ; SONG Yingxia 九州大学, 医学系学府
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ミスマッチ修復 / 細胞死 / DNA損傷 / 8-オキソグアニン / 酸化ストレス / 複合体

Outline of Final Research Achievements

We observed an oxidative stress-dependent complex formation of MUTYH and mismatch repair factors. Using anti-FLAG antibody, we purified the complex from FLAG-tagged MUTYH expressing human cells. Western blot analyses revealed that the complex contained DNA polymerase δ and MLH 1 in addition to MSH 2, MSH 6, and PCNA. The interaction between MUTYH and MSH2 was also observed in the 8-oxo-dGua administered cells, suggesting that 8-oxoG is involved in the formation of this complex. In cells loaded with oxidative stress, transient mono-ubiquitination of PCNA occurs, but PCNA in this complex has not undergone ubiquitination. Certain MSH2 mutants found in Lynch syndrome and a DNA polymerase δ mutant do not form the oxidative stress-dependent complexes with MUTYH. These results suggested that DNA polymerase δ as well as MSH2 might be involved in molecular mechanisms underlying oxidative stress induced cell-death.

Research Products

• [Journal Article] Celecoxib and 2,5-dimethylcelecoxib inhibit intestinal cancer growth by suppressing the Wnt/β-catenin signaling pathway (2017)
  • Author(s): I. Egashira, F. Takahashi-Yanaga, R. Nishida, M. Arioka, K. Igawa, K. Tomooka, Y. Nakatsu, T. Tsuzuki, Y. Nakabeppu, T. Kitazono, T. Sasaguri,
  • Journal Title: Cancer Science Volume: 108 Issue: 1 Pages: 108-115
  • DOI: 10.1111/cas.13106
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• [Journal Article] Insertion and deletion mismatches distant from the target position improve gene correction with a tailed duplex. (2016)
  • Author(s): H. Kamiya*, N. Nishigaki, A. Ikeda, S. Yukawa, Y. Morita, Y. Nakatsu, T. Tsuzuki and H. Harashima
  • Journal Title: Nucleosides Nucleotides Nucleic Acids Volume: 35 Issue: 7 Pages: 379-388
  • DOI: 10.1080/15257770.2016.1163384
  • NAID: 120006313423
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] BUBR1 insufficiency in mice increases their sensitivity to oxidative stress (2016)
  • Author(s): Matsuda, D., Matsumoto, T., Honma, K., Ikawa-Yoshida, A., Onimaru, M., Furuyama, T., Nakatsu, Y., Tsuzuki, T., Maehara, Y.
  • Journal Title: In Vvo Volume: 30 Pages: 769-776
  • Peer Reviewed / Open Access
• [Journal Article] BubR1 Insufficiency Impairs Liver Regeneration in Aged Mice after Hepatectomy through Intercalated Disc Abnormality (2016)
  • Author(s): A. Ikawa-Yoshida, T. Matsumoto, S. Okano, Y. Aoyagi, Y. Matsubara, T. Furuyama, Y. Nakatsu, T. Tsuzuki, M. Onimaru, T. Ohkusa, M. Nomura, Y. Maehara,
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 32399-32399
  • DOI: 10.1038/srep32399
  • Peer Reviewed / Open Access
• [Journal Article] BTK gene targeting by homologous recombination using a helper-dependent adenovirus/adeno-associated virus hybrid vector. (2016)
  • Author(s): Yamamoto H, Ishimura M, Ochiai M, Takada H, Kusuhara K, Nakatsu Y, Tsuzuki T, Mitani K, Hara T.
  • Journal Title: Gene Ther Volume: 23 Issue: 2 Pages: 205-13
  • DOI: 10.1038/gt.2015.91
  • Peer Reviewed / Open Access
• [Journal Article] Differentiation-inducing factor-3 inhibits intestinal tumor growth in vitro and in vivo. (2015)
  • Author(s): Kubokura N, Takahashi-Yanaga F, Arioka M, Yoshihara T, Igawa K, Tomooka K, Morimoto S, Nakatsu Y, Tsuzuki T, Nakabeppu Y, Matsumoto T, Kitazono T, Sasaguri T
  • Journal Title: J Pharmacol Sci Volume: 127 Issue: 4 Pages: 446-455
  • DOI: 10.1016/j.jphs.2015.03.005
  • Peer Reviewed / Open Access
• [Journal Article] UV-induced mutations in epidermal cells of mice defective in DNA polymerase η and/or ι (2015)
  • Author(s): Kanao R, Yokoi M, Ohkumo T, Sakurai Y, Dotsu K, Kura S, Nakatsu Y, Tsuzuki T, Masutani C, Hanaoka F
  • Journal Title: DNA Repair (Amsterdam) Volume: S1568-7864(15) Pages: 00042-7
  • DOI: 10.1016/j.dnarep.2015.02.006
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] BubR1 insufficiency inhibits neointimal hyperplasia through impaired vascular smooth muscle cell proliferation in mice (2015)
  • Author(s): Kyuragi, R., Matsumoto, T., Harada, Y., Saito, S., Onimaru, M., Nakatsu, Y., Tsuzuki, T., Nomura, M., Yonemitsu, Y., Maehara, Y.
  • Journal Title: Arterioscler. Thromb. Vasc. Biol. Volume: 35 Issue: 2 Pages: 341-347
  • DOI: 10.1161/atvbaha.114.304737
  • Peer Reviewed / Open Access
• [Journal Article] Mismatch repair deficient mice show susceptibility to oxidative stress-induced intestinal carcinogenesis (2014)
  • Author(s): J. Piao, Y. Nakatsu, M. Ohno, K. Taguchi, T. Tsuzuki
  • Journal Title: International Journal of Biological Sciences Volume: 10 Issue: 1 Pages: 73-79
  • DOI: 10.7150/ijbs.5750
  • Peer Reviewed / Open Access
• [Journal Article] DIF-1 inhibits tumor growth in vivo reducing phosphorylation of GSK-3beta and expressions of cyclin D1 and TCF7L2 in cancer model mice. (2014)
  • Author(s): Takahashi-Yanaga, F., Yoshihara, T., Jingushi, K., Igawa, K., Tomooka, K., Watanabe, Y., Morimoto, S., Nakatsu, Y., Tsuzuki, T., Nakabeppu, Y., and Sasaguri, T.
  • Journal Title: Biochem Pharmacol Volume: N/A Issue: 3 Pages: 340-348
  • DOI: 10.1016/j.bcp.2014.03.006
  • Peer Reviewed / Open Access
• [Journal Article] Abnormality in Wnt signaling is causatively associated with oxidative stress-induced intestinal tumorigenesis in MUTYH-null mice (2014)
  • Author(s): Isoda T, Nakatsu Y, Yamauchi K, Piao J, Yao T, Honda H, Nakabeppu Y, Tsuzuki T
  • Journal Title: Int J Biol Sci Volume: 10 Issue: 8 Pages: 940-947
  • DOI: 10.7150/ijbs.9241
  • Peer Reviewed / Open Access
• [Presentation] Mutyh欠損マウスにおける酸化ストレスによる消化管腫瘍発生頻度上昇と特異的体細胞変異シグニチャーの解析 (2016)
  • Author(s): 鷹野典子、大野みずき、中津可道、中別府雄作、續輝久
  • Organizer: 日本分子生物学会第39回年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-12-02
• [Presentation] ヒト細胞由来ミスマッチ修復遺伝子変異体の作製とその解析 (2016)
  • Author(s): 林田元気、中津可道、日高京子、藤兼亮輔、日高真純、釣本敏樹、續輝久
  • Organizer: 日本分子生物学会第39回年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-12-02
• [Presentation] ミスマッチ修復欠損マウスを用いた新規生殖細胞変異の検出 (2016)
  • Author(s): 大野みずき、鷹野典子、佐々木史子、中津可道、續輝久
  • Organizer: 日本環境変異原学会第45回大会
  • Place of Presentation: つくば
  • Year and Date: 2016-11-17
• [Presentation] Development of assay systems to characterize the variants of mismatch repair factor MSH2 found in Lynch syndrome (2016)
  • Author(s): Hayashida, G., Nakatsu, Y., Hidaka, K., Fujikane, R., Hidaka, M., Tsurimoto, T., Tsuzuki, T.
  • Organizer: The 10th International 3R (Replication, Recombination and Repair) Symposium
  • Place of Presentation: 松江
  • Year and Date: 2016-11-14
  • Int'l Joint Research
• [Presentation] Characterization of mismatch repair factor MSH2 variants found in Lynch syndrome (2016)
  • Author(s): 林田元気、中津可道、日高京子、藤兼亮輔、日高真純、釣本敏樹、續輝久
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: 広島
  • Year and Date: 2016-10-23
• [Presentation] 酸化ストレスによる消化管腫瘍発生頻度上昇と特異的体細胞変異シグニチャー： Mutyh欠損マウスを用いた解析 (2016)
  • Author(s): 大野みずき、鷹野典子、中津可道、中別府雄作、續輝久
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-07
• [Presentation] ヒト細胞を用いたミスマッチ修復因子MSH2の変異体の解析 (2016)
  • Author(s): 林田元気、中津可道、日高京子、藤兼亮輔、日高真純、續輝久
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-07
• [Presentation] CRISPR/Cas9を用いたヒト細胞のミスマッチ修復因子への変異導入 (2016)
  • Author(s): 林田元気、中津可道、日高京子、 藤兼亮輔、日高真純、釣本敏樹、 續輝久
  • Organizer: 日本遺伝学会第88回大会
  • Place of Presentation: 三島
  • Year and Date: 2016-09-09
• [Presentation] ミスマッチDNA修復系が体細胞および生殖細胞ゲノムにおよぼす影響 (2016)
  • Author(s): 鷹野典子、大野みずき、 佐々木史子、中津可道、續輝久
  • Organizer: 日本遺伝学会第88回大会
  • Place of Presentation: 三島
  • Year and Date: 2016-09-09
• [Presentation] ミスマッチ修復因子MSH2の発現系の確立と変異体の解析 (2015)
  • Author(s): 林田 元気、中津 可道、日高 京子、藤兼 亮輔、日高 真純、釣本 敏樹、續 輝久
  • Organizer: 日本分子生物学会第38回年会・日本生化学会第88回大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] Mutyh欠損マウスを用いた酸化ストレス誘発突然変異と発がんの解析 (2015)
  • Author(s): 鷹野典子、大野みずき、佐々木史子、山内一己、中別府雄作、中津可道、續輝久
  • Organizer: 日本分子生物学会第38回年会・日本生化学会第88回大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
• [Presentation] 酸化ストレス誘発マウス小腸発がん実験から見えてきたこと (2015)
  • Author(s): 中津可道
  • Organizer: 日本環境変異原学会第44回大会
  • Place of Presentation: 福岡
  • Year and Date: 2015-11-27
  • Invited
• [Presentation] ミスマッチ修復欠損マウスにおける生殖細胞ゲノム変異の解析 (2015)
  • Author(s): 大野みずき、鷹野典子、佐々木史子、日高京子、中津可道、續輝久
  • Organizer: 日本環境変異原学会第44回大会
  • Place of Presentation: 福岡
  • Year and Date: 2015-11-27
  • Invited
• [Presentation] 酸化ストレス誘発消化管発がんと突然変異の抑制におけるMUTYHの役割 (2015)
  • Author(s): 大野みずき、鷹野典子、中津可道、中別府雄作、續輝久
  • Organizer: 日本癌学会第74回学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2015-10-08
• [Presentation] Role of the oxidative DNA damage repair system in somatic and germline mutations in mice (2015)
  • Author(s): Mizuki Ohno, Noriko Takano, Kunihiko Sakumi, Ryutaro Fukumura, Yuki Iwasaki, Toshimichi Ikemura, Yoichi Gondo, Yusaku Nakabeppu, Yoshimichi Nakatsu, Teruhisa Tsuzuki
  • Organizer: Zing conference "Genome Integrity"
  • Place of Presentation: Cairns, Australia
  • Year and Date: 2015-07-31
  • Int'l Joint Research / Invited
• [Presentation] Mismatch Repair Deficient Mice Show Susceptibility to Oxidative Stress-induced Intestinal Carcinogenesis (2015)
  • Author(s): Yoshimich Nakatsu, Jingshu Piao, Takuya Hashizume, Mizuki Ohno, Kenichi Taguchi, Teruhisa Tsuzuki
  • Organizer: 15th International Congress of Radiation Research
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] Oxidative stress-induced intestinal tumors in Mutyh-deficient mice treated with low doses of potassium bromate (2014)
  • Author(s): Teruhisa Tsuzuki, Mizuki Ohno, Noriko Takano, Kenichi Taguchi, Yusaku Nakabeppu, Yasunobu Aoki, Takehiko Nohmi, Yoshimichi Nakatsu
  • Organizer: 4th Asian Conference on Environmental Mutagen
  • Place of Presentation: Kolkata, India
  • Year and Date: 2014-12-10 – 2014-12-12
  • Invited
• [Presentation] Mutyh遺伝子欠損マウスを用いた酸化ストレス誘発消化管がんの解析 (2014)
  • Author(s): 大野みずき、鷹野典子、佐々木史子、田口健一、中別府雄作、青木康展、能美健彦、中津可道、續輝久
  • Organizer: 日本環境変異原学会第43回大会
  • Place of Presentation: 東京
  • Year and Date: 2014-12-04 – 2014-12-05
• [Presentation] Mutyh遺伝子欠損マウスにおける酸化ストレス誘発突然変異の解析 (2014)
  • Author(s): 鷹野典子、大野みずき、稲葉洋平、志村勉、欅田尚樹、中別府雄作、中津可道、續輝久
  • Organizer: 日本環境変異原学会第43回大会
  • Place of Presentation: 東京
  • Year and Date: 2014-12-04 – 2014-12-05
• [Presentation] Mutyh遺伝子欠損マウスにおける酸化ストレス誘発突然変異の解析 (2014)
  • Author(s): 鷹野典子、大野みずき、稲葉洋平、志村勉、欅田尚樹、中別府雄作、中津可道、續輝久
  • Organizer: 日本分子生物学会第37回年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 非標的部位におけるミスマッチの配列変換（遺伝子修復）への影響 (2014)
  • Author(s): 西垣奈津希、池田彰弘、湯川誠也、守田由子、中津可道、續輝久、原島秀吉、紙谷浩之
  • Organizer: 日本分子生物学会第37回年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Oxidative stress-induced intestinal tumors in Mutyh-deficient mice treated with low doses of potassium bromate (2014)
  • Author(s): Teruhisa Tsuzuki, Mizuki Ohno, Noriko Takano, Kenichi Taguchi, Yusaku Nakabeppu, Yasunobu Aoki, Takehiko Nohmi, Yoshimichi Nakatsu
  • Organizer: 5th US-Japan DNA Repair Meeting
  • Place of Presentation: Tokushima, Japan
  • Year and Date: 2014-10-28 – 2014-10-31
  • Invited
• [Presentation] 酸化ストレス誘発突然変異と消化管がん解析 (2014)
  • Author(s): 大野みずき、鷹野典子、佐々木史子、橋詰拓弥、李賛、田口健一、中別府雄作、中津可道、續輝久
  • Organizer: 日本放射線影響学会第57回大会
  • Place of Presentation: 鹿児島
  • Year and Date: 2014-10-01 – 2014-10-03
• [Presentation] ミスマッチ修復欠損マウスにおける酸化ストレス誘発突然変異の解析 (2014)
  • Author(s): 橋詰拓弥、中津可道、大野みずき、佐々木史子、鷹野典子、續輝久
  • Organizer: 日本放射線影響学会第57回大会
  • Place of Presentation: 鹿児島
  • Year and Date: 2014-10-01 – 2014-10-03
• [Presentation] DIF-1はGSK-3βのリン酸化抑制とcyclin D1 および TCF7L2の発現低下により抗腫瘍効果を発揮する (2014)
  • Author(s): 高橋富美、中津可道、續輝久、中別府雄作、笹栗俊之
  • Organizer: 日本癌学会第73回学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] マイクロサテライト不安定性は変異導入を伴ってDNA 複製ストレス下で誘導される (2014)
  • Author(s): 熱海悠子、片山琴絵、島村徹平、中津可道、益谷美都子、宮野悟、中釜斉、續輝久、吉岡研一
  • Organizer: 日本癌学会第73回学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] 低用量臭素酸カリウムの飲水投与によりMutyh遺伝子欠損マウスの消化管で誘発された突然変異並びに発がんの解析 (2014)
  • Author(s): 大野みずき、鷹野典子、中津可道、中別府雄作、續輝久
  • Organizer: 日本癌学会第73回学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Mutyh欠損マウスにおける酸化ストレス誘発発がんおよび突然変異の解析 (2014)
  • Author(s): 鷹野典子, 大野みずき, 中別府雄作, 中津可道, 續輝久
  • Organizer: 日本遺伝学会第86回大会
  • Place of Presentation: 長浜
  • Year and Date: 2014-09-17 – 2014-09-19
• [Book] 日本臨牀（増刊号）最新臨床大腸癌学 基礎研究から臨床応用へ (Ⅲ－２－(6))酸化DNA損傷と大腸発癌） (2015)
  • Author(s): 續輝久、大野みずき、中津可道
  • Total Pages: 715
  • Publisher: 株式会社 日本臨牀社