Biochemical study on the molecular mechanisms of induced mutagenesis and post-replication repair pathways

• Project/Area Number: 24310040
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Risk sciences of radiation/Chemicals
• Research Institution: Nagoya University
• Principal Investigator: Masuda Yuji 名古屋大学, 医学系研究科(環医), 准教授 (30273866)
• Co-Investigator(Renkei-kenkyūsha): MASUTANI Chikahide 名古屋大学, 環境医学研究所, 教授 (40241252) ; Kanao Rie 名古屋大学, 環境医学研究所, 助教 (30542287)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥13,130,000 (Direct Cost: ¥10,100,000、Indirect Cost: ¥3,030,000); Fiscal Year 2014: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2013: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2012: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: 複製後修復 / 損傷乗り越え DNA 合成 / 脱ユビキチン / PCNA / ポリユビキチン化 / 突然変異誘発 / 損傷乗り越えDNA合成 / ユビキチン / HLTF / RAD6-RAD18 / template switch

Outline of Final Research Achievements

Radiation and environmental mutagens induce mutations. To elucidate molecular mechanisms of the induced mutagenesis is one of the biggest issues in this field. A large fraction of the induced mutation is generated by a cellular process, post-replication repair pathway. In humans, cells have two sub-pathways. One is the error-prone pathway, translesion DNA synthesis (TLS). The other is the error-free, in principle, pathway, template switch (TS). The regulation of the pathway choice that is a crucial step for the maintenance of genetic stability is regulated by ubiquitination of PCNA. In this study, we established in vitro reconstitution systems for PCNA ubiquitination and deubiquitination, and analyzed molecular mechanisms of these biochemical reactions.

Research Products

• [Journal Article] Relevance of simultaneous mono-ubiquitinations of multiple units of PCNA homo-trimers in DNA damage tolerance. (2015)
  • Author(s): Kanao R, Masuda Y, Deguchi S, Yumoto-Sugimoto M, Hanaoka F, Masutani C
  • Journal Title: PLoS One Volume: 10 Issue: 2 Pages: e0118775-e0118775
  • DOI: 10.1371/journal.pone.0118775
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Guanine-5-carboxylcytosine base pairs mimic mismatches during DNA replication. (2014)
  • Author(s): Shibutani T, Ito S, Toda M, Kanao R, Collins LB, Shibata M, Urabe M, Koseki H, Masuda Y, Swenberg JA, Masutani C, Hanaoka F, Iwai S, Kuraoka I
  • Journal Title: Scientific Reports Volume: 4 Issue: 1 Pages: 5220-5220
  • DOI: 10.1038/srep05220
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A novel ATM/TP53/p21-mediated checkpoint only activated by chronic γ-irradiation (2014)
  • Author(s): CAO Lili, KAWAI Hidehiko, SASATANI Megumi, IIZUKA Daisuke, MASUDA Yuji, INABA Toshiya, SUZUKI Keiji, OOTSUYAMA Akira, UMATA Toshiyuki, KAMIYA Kenji, SUZUKI Fumio
  • Journal Title: PLoS One Volume: 9 Issue: 8 Pages: e104279-e104279
  • DOI: 10.1371/journal.pone.0104279
  • NAID: 120006986457
  • Peer Reviewed / Open Access
• [Journal Article] Strand breakage of (6-4) photoproduct-containing DNA at neutral pH and its repair by the ERCC1-XPF protein complex (2013)
  • Author(s): Arichi N, Yamamoto J, Takahata C, Masuda Y, Kuraoka I, Iwai S
  • Journal Title: Org. Biomol. Chem. Volume: 11 Issue: 21 Pages: 3526-3534
  • DOI: 10.1039/c3ob00012e
  • Peer Reviewed
• [Journal Article] A novel interplay between the Fanconi anemia core complex and ATR-ATRIP kinase during DNA cross-link repair (2013)
  • Author(s): J Tomida, A Itaya, T Shigechi, J Unno, E Uchida, M Ikura, Y Masuda, S Matsuda J Adachi, M Kobayashi, AR Meetei, Y Maehara, K Yamamoto K Kamiya, A Matsuura, T Matsuda. T Ikura, M Ishiai, M Takata
  • Journal Title: Nucleic Acids Research Volume: VOL. 41 Issue: 14 Pages: 6930-6941
  • DOI: 10.1093/nar/gkt467
  • Peer Reviewed
• [Journal Article] <i>In Vitro</i> Studies of Exchanges between Replicative and Translesion DNA Polymerases in the Eukaryotic Post-replication Repair Pathway (2012)
  • Author(s): Masuda Y*
  • Journal Title: Genes and Environment Volume: 34 Issue: 2 Pages: 70-76
  • DOI: 10.3123/jemsge.34.70
  • NAID: 10030311492
  • ISSN: 1880-7046, 1880-7062
  • Peer Reviewed
• [Journal Article] En bloc transfer of polyubiquitin chains to PCNA in vitro is mediated by two different human E2-E3 pairs (2012)
  • Author(s): Masuda Y
  • Journal Title: Nucleic Acids Res. Volume: 40(20) Issue: 20 Pages: 10394-10407
  • DOI: 10.1093/nar/gks763
  • Peer Reviewed
• [Presentation] ヒト細胞におけるPCNAの翻訳後修飾によるDNA損傷トレランス制御機構の解析 (2015)
  • Author(s): 金尾梨絵, 増田雄司, 花岡文雄, 益谷央豪
  • Organizer: 平成26年度「がん研究分野の特性等を踏まえた支援活動」公開シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2015-01-27 – 2015-01-28
• [Presentation] HECTタイプのユビキチンリガーゼ、E6AP-E6複合体によるユビキチン鎖伸長反応の分子機構 (2014)
  • Author(s): 増田雄司, 柊元 巌, 益谷央豪
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 変異体PCNA発現ヒト細胞を用いたDNA損傷トレランス制御の解析 (2014)
  • Author(s): 金尾梨絵, 増田雄司, 花岡文雄, 益谷央豪
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Interactions between PCNA and Y-family DNA polymerases (2014)
  • Author(s): Masuda Y, Kanao R, Ohmori H, Hanaoka F, Masutani C
  • Organizer: The 9th 3R (Replication, Recombination and Repair) Symposium
  • Place of Presentation: 御殿場
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] Relevance of simultaneous mono-ubiquitinations of multiple units of PCNA homo-trimers in DNA damage tolerance (2014)
  • Author(s): Masutani C, Kanao R, Masuda Y, Hanaoka F
  • Organizer: The 9th 3R (Replication, Recombination and Repair) Symposium
  • Place of Presentation: 御殿場
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] Analysis of a novel DNA-dependent ubiquitin ligase, human HLTF (2014)
  • Author(s): Matsumoto S, Masuda Y, Masutani C
  • Organizer: The 9th 3R (Replication, Recombination and Repair) Symposium
  • Place of Presentation: 御殿場
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] ユビキチンリガーゼによるユビキチン鎖リンケージとユビキチン鎖の伸長制御 (2014)
  • Author(s): 増田雄司
  • Organizer: 国立遺伝学研究所研究集会「染色体ＤＮＡの安定維持の分子メカニズム」
  • Place of Presentation: 三島
  • Year and Date: 2014-11-06 – 2014-11-07
  • Invited
• [Presentation] DNAポリメラーゼηとPCNAの相互作用の解析 (2014)
  • Author(s): 増田雄司, 金尾梨絵, 大森治夫, 花岡文雄, 益谷央豪
  • 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] DNAポリメラーゼηのPCNA相互作用部位の解析 (2014)
  • Author(s): 増田雄司, 金尾梨絵, 大森治夫, 花岡文雄, 益谷央豪
  • 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
• [Presentation] Different types of interactions between PCNA and Y-family DNA polymerases for translesion DNA synthesis (2014)
  • Author(s): Masud Y, Kanao R, Ohmori H, Hanaoka F, Masutani C
  • Organizer: ZING conference, DNA Polymerases: Biology, Diseases and Biomedical Applications Conference 2014
  • Place of Presentation: Cambridge, UK
  • Year and Date: 2014-08-31 – 2014-09-04
• [Presentation] The short CCG run in Huntingtin (HTT) gene is an obstacle for replicative DNA polymerases, potentially hampering the fork progression (2014)
  • Author(s): Hang LP, Furukohri A, Masuda Y, Maki S, Katayama T, Maki H
  • Organizer: ZING conference, DNA Polymerases: Biology, Diseases and Biomedical Applications Conference 2014
  • Place of Presentation: Cambridge, UK
  • Year and Date: 2014-08-31 – 2014-09-04
• [Presentation] PCNAのユビキチン修飾による複製後修復経路の制御 (2014)
  • Author(s): 増田雄司, 益谷央豪
  • Organizer: 変異機構研究会・第27回夏の学校
  • Place of Presentation: 小牧
  • Year and Date: 2014-06-21 – 2014-06-22
• [Presentation] Two different types of interactions between PCNA and Y-family DNA polymerases have distinct roles for translesion DNA synthesis (2014)
  • Author(s): Y Masuda, R Kanao, H Ohmori, F Hanaoka, C Masutani
  • Organizer: International Symposium on Xeroderma Pigmentosum and Related Diseases: Disorders of DNA Damage Response-Bench to Bedside-
  • Place of Presentation: 神戸国際会議場（神戸）
• [Presentation] Yファミリ―DNAポリメラーゼとPCNAとの相互作用による損傷乗り越えDNA合成経路の制御 (2013)
  • Author(s): 増田雄司, 金尾梨絵, 益谷央豪
  • Organizer: 変異機構研究会・第26回夏の学校
  • Place of Presentation: アイプラザ一宮（一宮）
• [Presentation] Yファミリ―DNAポリメラーゼとPCNAとの相互作用の解析 (2013)
  • Author(s): 増田雄司, 金尾梨絵, 益谷央豪
  • Organizer: 日本遺伝学会第85回大会
  • Place of Presentation: 慶應義塾大学 日吉キャンパス（横浜）
• [Presentation] DNAポリメラーゼηとPCNAとの相互作用による損傷乗り越えDNA合成経路の制御 (2013)
  • Author(s): 増田雄司, 金尾梨絵, 益谷央豪
  • Organizer: 日本放射線影響学会第56回大会
  • Place of Presentation: ホテルクラウンパレス青森 （青森）
• [Presentation] ヒトYファミリ―DNAポリメラーゼとPCNAとの相互作用 (2013)
  • Author(s): 増田雄司, 金尾梨絵, 益谷央豪
  • Organizer: 第22回DNA複製・組換え・修復ワークショップ
  • Place of Presentation: ホテルニュー水戸屋（仙台）
• [Presentation] PCNAのユビキチン修飾による複製後修復経路の制御 (2013)
  • Author(s): 増田雄司
  • Organizer: 国立遺伝学研究所・研究会「染色体DNAの安定維持の分子メカニズム」
  • Place of Presentation: 国立遺伝学研究所（三島）
  • Invited
• [Presentation] En bloc transfer of poly-ubiquitin chains to PCNA in vitro, implication in regulation of post-replication repair pathway (2012)
  • Author(s): Masuda Y, Suzuki M, Kawai H, Hishiki A, Hashimoto H, Masutani, C, Hishida T, Suzuki F, Kamiya K
  • Organizer: The 28th Radiation Biology Center International Symposium
  • Place of Presentation: COOP Inn Kyoto (京都)
  • Year and Date: 2012-11-29
• [Presentation] En bloc transfer of poly-ubiquitin chains to PCNA in vitro is mediated by two different human E2-E3 pairs (2012)
  • Author(s): Masuda Y, Suzuki M, Kawai H, Hishiki A, Hashimoto H, Masutani C, Hishida T, Suzuki F, Kamiya K
  • Organizer: The 8th 3R (Replication, Recombination and Repair) Symposium
  • Place of Presentation: 淡路夢舞台国際会議場(淡路)
  • Year and Date: 2012-11-26
• [Presentation] PCNAのユビキチン化の分子機構と複製後修復経路の制御 (2012)
  • Author(s): 増田雄司
  • Organizer: 日本環境変異原学会-変異機構研究会・第25回夏の学校
  • Place of Presentation: 小牧勤労センター(小牧)
• [Presentation] En bloc transfer of poly-ubiquitin chains to PCNA in vitro mediated by two human E2-E3 pairs (2012)
  • Author(s): Masuda Y
  • Organizer: Mutagenesis, Gordon Research Conferences, A Delicate Balance : Cellular Mutation Pathways in Genetic Stability and Disease
  • Place of Presentation: Salve Regina University (Newport, USA)
• [Presentation] ヒトPCNAのユビキチン化酵素RAD6-RAD18複合体の構造と機能 (2012)
  • Author(s): 増田雄司、鈴木美紀、益谷央豪、神谷研二
  • Organizer: 日本放射線影響学会第55回大会
  • Place of Presentation: 東北大学川内北キャンパス(仙台)
• [Presentation] PCNAのユビキチン化の新規分子機構 (2012)
  • Author(s): 増田雄司、益谷央豪
  • Organizer: 日本遺伝学会第84回大会
  • Place of Presentation: 九州大学医学部(福岡)(招待講演)
• [Presentation] ヒトPCNAのポリュビキチン化の新規分子機構 (2012)
  • Author(s): 増田雄司、鈴木美紀、河合秀彦、菱木麻美、橋本 博、益谷央豪、神谷研二
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場(福岡)
• [Remarks] 名古屋大学環境医学研究所
  • URL: http://www.riem.nagoya-u.ac.jp
• [Remarks] 名古屋大学環境医学研究所ゲノム動態制御分野
  • URL: http://www.riem.nagoya-u.ac.jp/4/genome/home.html
• [Remarks] 名古屋大学環境医学研究所
  • URL: http://www.riem.nagoya-u.ac.jp
• [Remarks] 名古屋大学環境医学研究所ゲノム動態制御分野
  • URL: http://www.riem.nagoya-u.ac.jp/4/genome/home.html
• [Remarks]
  • URL: http://www.riem.nagoya-u.ac.jp/