Rescue of arrested replication forks by multiple ubiquitination of PCNA

• Project/Area Number: 26740017
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Nagoya University
• Principal Investigator: Kanao Rie 名古屋大学, 環境医学研究所, 助教 (30542287)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: DNA損傷トレランス / PCNA / モノユビキチン化 / 損傷乗り越え複製 / 翻訳後修飾

Outline of Final Research Achievements

DNA damage tolerance is an important mechanism to prevent DNA replication arrest by DNA damage. It is suggested that DNA damage tolerance is regulated by post-translational modifications of PCNA at K164. PCNA forms a ring-shaped structure consist of homo-trimeric PCNA molecules, which means a single PCNA ring have three modifiable K164. In this study, I investigated the mechanism of DNA damage tolerance pathway which is regulated by simultaneously mono-ubiquitinated PCNA on three K164 in a ring. I analyzed the responses of the cells expressing ectopic PCNA mutated at K164 to DNA damaging agents.

Research Products

• [Journal Article] Different types of interaction between PCNA and PIP boxes contribute to distinct cellular functions of Y-family DNA polymerases. (2015)
  • Author(s): Masuda Y, Kanao R, Kaji K, Ohmori H, Hanaoka F, Masutani C
  • Journal Title: Nucleic Acids Res Volume: 43 Issue: 16 Pages: 7898-7910
  • DOI: 10.1093/nar/gkv712
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] USP7 is a suppressor of PCNA ubiquitination and oxidative-stress-induced mutagenesis in human cells. (2015)
  • Author(s): Kashiwaba S, Kanao R, Masuda Y, Kusumoto-Matsuo R, Hanaoka F, Masutani C
  • Journal Title: Cell Rep Volume: 13 Issue: 10 Pages: 2072-2080
  • DOI: 10.1016/j.celrep.2015.11.014
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [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] 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] 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
• [Presentation] PCNAの翻訳後修飾が制御するDNA損傷トレランス機構 (2017)
  • Author(s): 金尾梨絵、 増田雄司、 益谷央豪
  • Organizer: 日本薬学会第137年会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-24
• [Presentation] PCNAの翻訳後修飾が制御するDNA損傷トレランスの解析 (2016)
  • Author(s): 金尾梨絵、 増田雄司、 益谷央豪
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-11-30
• [Presentation] Suppression of PCNA ubiquitination and oxidative stress-induced mutagenesis by USP7 (2016)
  • Author(s): Kanao R, Kashiwaba S, Masuda Y, Kusumoto-Matsuo R, Hanaoka F, Masutani C
  • Organizer: The 10th 3R (Replication, Recombination and Repair) symposium
  • Place of Presentation: Matsue
  • Year and Date: 2016-11-13
  • Int'l Joint Research
• [Presentation] PCNAのモノユビキチン化の制御によるヒト細胞での酸化的DNA損傷誘発突然変異抑制機構 (2016)
  • Author(s): 金尾梨絵、柏葉脩一郎、増田雄司、松尾（楠本）理加、花岡文雄、益谷央豪
  • Organizer: 日本薬学会 第136年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-03-26
• [Presentation] PCNAのユビキチン化を制御して過酸化水素誘発突然変異を抑制するヒト細胞のメカニズム (2015)
  • Author(s): 益谷央豪、柏葉脩一郎、金尾梨絵、松尾（楠本）理加、花岡文雄、増田雄司
  • Organizer: 第38回日本分子生物学会年会第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] PCNAとヒトDNAポリメラーゼηの相互作用 (2015)
  • Author(s): 増田雄司、金尾梨絵、益谷央豪
  • Organizer: 日本環境変異原学会第44回大会
  • Place of Presentation: 九州大学
  • Year and Date: 2015-11-27
• [Presentation] 酸化損傷によって誘導されるPCNAのモノユビキチン化を制御する新規メカニズムの解析 (2015)
  • Author(s): 金尾梨絵、柏葉脩一郎、増田雄司、松尾（楠本）理加、花岡文雄、益谷央豪
  • Organizer: 第23回DNA複製・組換え・修復ワークショップ
  • Place of Presentation: 焼津
  • Year and Date: 2015-10-19
• [Presentation] PCNAによるヒトDNAポリメラーゼηの活性促進 (2015)
  • Author(s): 増田雄司、金尾梨絵、益谷央豪
  • Organizer: 第23回DNA複製・組換え・修復ワークショップ
  • Place of Presentation: 焼津
  • Year and Date: 2015-10-19
• [Presentation] Regulation of DNA damage tolerance by simultaneous mono-ubiquitinations of multiple-units of PCNA in human cells. (2015)
  • Author(s): Kanao R, Masuda Y, Hanaoka F, Masutani C
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2015-10-08
• [Presentation] PCNAによるヒトDNAポリメラーゼηの活性促進機構 (2015)
  • Author(s): 増田雄司、金尾梨絵、益谷央豪
  • Organizer: 日本遺伝学会第87回大会
  • Place of Presentation: 仙台
  • Year and Date: 2015-09-24
• [Presentation] Disruption of Simultaneous Mono-ubiquitinations of Multiple Units of PCNA Homo-trimers Disturbs DNA Damage Tolerance in Human Cells. (2015)
  • Author(s): Masutani C, Kanao R, Masuda Y, Hanaoka F
  • Organizer: ICRR 2015 15th International Congress of Radiation Resarch
  • Place of Presentation: 京都
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [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] 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] 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] Relevance of simultaneous mono-ubiquitinations of multiple units of PCNA homo-trimers in DNA damage torelance (2014)
  • Author(s): Masutani C, Kanao R, Masuda Y, Hanaoka F
  • Organizer: US-Japan DNA Repair Meeting (日米修復会議)
  • Place of Presentation: 徳島
  • Year and Date: 2014-10-28 – 2014-10-31
  • Invited
• [Presentation] 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 (2014)
  • Author(s): 益谷央豪, 金尾梨絵, 増田雄司, 花岡文雄
  • 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
  • Invited