Novel molecular mechanism maintaining genomic homeostasis against environmental stresses

• Project/Area Number: 24241019
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Risk sciences of radiation/Chemicals
• Research Institution: Kobe University
• Principal Investigator: SUGASAWA Kaoru 神戸大学, 自然科学系先端融合研究環バイオシグナル研究センター, 教授 (70202124)
• Co-Investigator(Kenkyū-buntansha): IWAI Shigenori 大阪大学, 基礎工学研究科, 教授 (10168544)
• Co-Investigator(Renkei-kenkyūsha): YOKOTA Hiroaki 光産業創成大学院大学, 光産業創成研究科, 准教授 (90415547) ; YOSHINO Ken-ichi 神戸大学, 自然科学系先端融合研究環バイオシグナル研究センター, 助教 (90280792)
• Research Collaborator: AKITA Masaki ; MATSUMOTO Syota ; TONE Daisuke ; ONISHI Yuki ; KINOSHITA Minoru
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥43,030,000 (Direct Cost: ¥33,100,000、Indirect Cost: ¥9,930,000); Fiscal Year 2014: ¥13,260,000 (Direct Cost: ¥10,200,000、Indirect Cost: ¥3,060,000); Fiscal Year 2013: ¥13,130,000 (Direct Cost: ¥10,100,000、Indirect Cost: ¥3,030,000); Fiscal Year 2012: ¥16,640,000 (Direct Cost: ¥12,800,000、Indirect Cost: ¥3,840,000)
• Keywords: 遺伝子 / ゲノム / 蛋白質 / DNA損傷修復 / 一分子解析

Outline of Final Research Achievements

This research project was aimed to understand the molecular bases, which enable the global genomic nucleotide excision repair (GG-NER) pathway to efficiently survey genomic DNA for damage caused by various environmental stresses. By using the defined cell-free GG-NER system reconstituted with purified recombinant protein factors, it was shown that certain endogenous DNA lesions, such as abasic sites, facilitate the damage recognition process in GG-NER, and that DNA repair efficiency is affected by the topological state of damaged DNA as well as protein phosphorylation. We also elucidated novel roles of ubiquitination and SUMOylation of the xeroderma pigmentosum group C (XPC) protein in recognition of ultraviolet (UV)-induced DNA photolesions mediated by the UV-damaged DNA binding protein complex. Finally, single molecule imaging experiments revealed one-dimensional diffusion of the XPC protein during the search for lesions along DNA.

Research Products

• [Journal Article] Functional regulation of the DNA damage-recognition factor DDB2 by ubiquitination and interaction with xeroderma pigmentosum group C protein. (2015)
  • Author(s): Syota Matsumoto, Eric S. Fischer, Takeshi Yasuda, Naoshi Dohmae, Shigenori Iwai, Toshio Mori, Ryotaro Nishi, Ken-ichi Yoshino, Wataru Sakai, Fumio Hanaoka, Nicolas H. Thoma, Kaoru Sugasawa
  • Journal Title: Nucleic Acids Res. Volume: 3 Pages: 1700-1713
  • DOI: 10.1093/nar/gkv038
  • NAID: 120005621885
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Fluorescence detection of cellular nucleotide excision repair of damaged DNA. (2014)
  • Author(s): Toga, T., Kuraoka, I., Watanabe, S., Nakano, E., Takeuchi, S., Nishigori, C., Sugasawa, K. and Iwai, S.
  • Journal Title: Sci Rep. Volume: 4 Pages: 5578-5578
  • DOI: 10.1038/srep05578
  • Peer Reviewed / Open Access
• [Journal Article] PRDM14 promotes active DNA demethylation through the Ten-eleven translocation (TET)-mediated base excision repair pathway in embryonic stem cells. (2014)
  • Author(s): Okashita, N., Kumaki, Y., Ebi, K., Nishi, M., Okamoto, Y., Nakayama, M., Hashimoto, S., Nakamura, T., Sugasawa, K., Kojima, N., Takada, T., Okano, M. and Seki, Y.
  • Journal Title: Development Volume: 141 Pages: 269-280
  • DOI: 10.1242/dev.099622
  • Peer Reviewed
• [Journal Article] Comparative analysis of interaction of human and yeast DNA damage recognition complexes with damaged DNA in nucleotide excision repair. (2013)
  • Author(s): Krasikova, Y. S., Rechkunova, N. I., Maltseva, E. A., Pestryakov, P. E., Petruseva, I. O., Sugasawa, K., Chen, X., Min, J. H. and Lavrik, O. I.
  • Journal Title: J. Biol. Chem. Volume: 288 Pages: 10936-10947
  • DOI: 10.1074/jbc.m112.444026
  • Peer Reviewed
• [Journal Article] Structure-function analysis of the EF-hand protein centrin-2 for its intracellular localization and nucleotide excision repair. (2013)
  • Author(s): Nishi R, Sakai W, Tone D, Hanaoka F, Sugasawa K
  • Journal Title: Nucleic Acids Res. Volume: 41 Pages: 4362-4371
  • DOI: 10.1093/nar/gkt434
  • NAID: 120005621886
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Structure-specific endonucleases Xpf and Mus81 play overlapping but essential roles in DNA repair by homologous recombination. (2013)
  • Author(s): Kikuchi K, Narita T, Pham 〓, Iij〓 J, Hirota K, Islam SK, Mohiuddin M, Okawa K, Bori T, Fukagama T, Essers J, Kanser R, Whithy NC, Sugasawa K, Taniguchi T, Kitagawa K. Takeda S.
  • Journal Title: Cancer Research Volume: (in press) Pages: 4362-4371
  • DOI: 10.1158/0008-5472.can-12-3154
  • Peer Reviewed
• [Journal Article] Human and yeast DNA damage recognition complexes bind with high affinity DNA structures mimicking in size transcription bubble. (2013)
  • Author(s): Krasikova, Y. S., Rechkunova, N. I., Maltseva, E. A., Anarbaev, R. O., Pestryakov, P. E., Sugasawa, K., Min, J. H. and Lavrik, O. I.
  • Journal Title: Journal of Molecular Recognition Volume: 26 Pages: 653-661
  • DOI: 10.1002/jmr.2308
  • Peer Reviewed
• [Journal Article] PARP1 promotes nucleotide excision repair through DDB2 stabilization and recruitment of ALC1 (2012)
  • Author(s): Pines, A. et al.
  • Journal Title: J. Cell Biol. Volume: 199 Pages: 235-249
  • DOI: 10.1083/jcb.201112132
  • Peer Reviewed
• [Journal Article] Two different RFC proteins, Ctf18 and RFC1 separately control PCNA-CRL4Cdt2-mediated Cdt1 proteolysis during S phase and following UV-irradiation (2012)
  • Author(s): Shiomi Y, Ishii T, Hayashi A, Shinmyozu K, Nakayama J, Sugasawa K, & Nishitani H
  • Journal Title: Mol. Cell. Biol. Volume: 32 Pages: 2279-2288
  • DOI: 10.1128/mcb.06506-11
  • Peer Reviewed
• [Presentation] In vivo regulation of mammalian nucleotide excision repair. (2015)
  • Author(s): Sugasawa, K.
  • Organizer: Gordon Research Conference on Mammalian DNA Repair
  • Place of Presentation: Four Points Sheraton, Ventura, CA, U.S.A.
  • Year and Date: 2015-02-08 – 2015-02-14
  • Invited
• [Presentation] 哺乳類ヌクレオチド除去修復タンパク質XPCのDNA結合モードの1分子イメージング (2014)
  • Author(s): 横田浩章、戸根大輔、大西優貴、韓 龍雲、原田慶恵、菅澤 薫
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] ヌクレオチド除去修復におけるDNA損傷認識機構の解析 (2014)
  • Author(s): 大西優貴、戸根大輔、木下 実、岩井成憲、菅澤 薫
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] DNA損傷応答におけるDDB2の翻訳後修飾の機能解析 (2014)
  • Author(s): 松本翔太、Eric S. Fischer、Nicolas H. Thoma、菅澤 薫
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Functional interactions between XPC and DDB2 in nucleotide excision repair (2014)
  • Author(s): Matsumoto, S., Fischer, E. S., Thoma, N. H. and Sugasawa, K.
  • Organizer: The 9th 3R Symposium
  • Place of Presentation: 御殿場高原ホテル
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] ヌクレオチド除去修復におけるXPCとDDB2の機能的相互作用 (2014)
  • Author(s): 松本翔太、Eric S. Fischer、Nicolas H. Thoma、菅澤 薫
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] 紫外線誘発DNA損傷修復の細胞内制御機構 (2014)
  • Author(s): 菅澤 薫
  • Organizer: 日本放射線影響学会第57回大会
  • Place of Presentation: かごしま県民交流センター
  • Year and Date: 2014-10-01 – 2014-10-03
• [Presentation] Roles for post-translational protein modifications in regulation of DNA damage recognition for mammalian nucleotide excision repair. (2014)
  • Author(s): Akita, M., Matsumoto, S., Sakai, W. and Sugasawa, K.
  • Organizer: International Symposium on Xeroderma Pigmentosum and Related Diseases
  • Place of Presentation: 神戸国際会議場
• [Presentation] ゲノムDNA損傷の修復と細胞応答を制御するタンパク質翻訳後修飾の役割 (2013)
  • Author(s): 菅澤 薫
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 紫外線誘発DNA損傷の修復機構におけるSUMO化修飾の役割 (2013)
  • Author(s): 秋田眞季、酒井 恒、菅澤 薫
  • 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
• [Presentation] 哺乳類ヌクレオチド除去修復タンパク質XPCの1分子イメージング (2013)
  • Author(s): 横田浩章、西幹栄美、戸根大輔、韓 龍雲、原田慶恵、菅澤 薫
  • Organizer: 第51回日本生物物理学会年会
  • Place of Presentation: 国立京都国際会館
• [Presentation] ヌクレオチド除去修復における損傷認識機構とSUMO化修飾の意義 (2013)
  • Author(s): 秋田眞季、松本翔太、菅澤 薫
  • 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] 哺乳類ヌクレオチド除去修復におけるDNA損傷認識課程の制御 (2013)
  • Author(s): 菅澤 薫
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド
• [Presentation] 哺乳類ヌクレオチド除去修復タンパク質XPCの１分子イメージング：DNA上での１次元自由拡散運動の観察 (2013)
  • Author(s): 横田浩章、戸根大輔、韓 龍雲、原田慶恵、菅澤 薫
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド
• [Presentation] Post-translational protein modification regulating mammalian nucleotide excision repair (2012)
  • Author(s): Sugasawa, K.
  • Organizer: The 4th US-Japan DNA Repair Meeting
  • Place of Presentation: Leesburg, VA, USA
  • Invited
• [Presentation] 紫外線誘発DNA損傷の認識と修復の分子基盤 (2012)
  • Author(s): 菅澤 薫
  • Organizer: 第34回日本光医学・光生物学会
  • Place of Presentation: 神戸
  • Invited
• [Presentation] 紫外線によるDNA損傷の修復を促進するDDB2の構造と機能制御 (2012)
  • Author(s): 松本翔太 他
  • Organizer: 日本放射線影響学会第55回大会
  • Place of Presentation: 仙台
• [Presentation] Functional regulation of the DNA damage recognition factor DDB2 via post- translational modifications (2012)
  • Author(s): Sugasawa, K. and Hanaoka, F.
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: 札幌
• [Presentation] A novel function of centrin-2 involved in maintenance of genomic stability (2012)
  • Author(s): Sugasawa, K. et al.
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
• [Presentation] Exploration of novel regulatory factors in mammalian nucleotide excision repair (2012)
  • Author(s): Tone, D. et al.
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
• [Remarks] 神戸大学バイオシグナル研究センター 菅澤研究室
  • URL: http://www.research.kobe-u.ac.jp/brce-sugasawa/
• [Remarks] 神戸大学バイオシグナル研究センター 菅澤研究室
  • URL: http://www.research.kobe-u.ac.jp/brce-sugasawa/
• [Remarks] 神戸大学バイオシグナル研究センター 菅澤研究室
  • URL: http://www.research.kobe-u.ac.jp/brce-sugasawa/