Studies on novel molecular mechanisms that regulate repair of genomic DNA damage caused by environmental stresses

2011 Fiscal Year Final Research Report

• Project/Area Number: 20241013
• 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(Renkei-kenkyūsha): NISHI Ryotaro 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 (80446525)
• Project Period (FY): 2008 – 2011
• Keywords: ヌクレオチド除去修復 / XPC / UV-DDB / ユビキチン化 / SUMO化 / プロテアソーム / 紫外線

Research Abstract

This study has been aimed to elucidate novel molecular mechanisms involved in spatio-temporal regulation of mammalian nucleotide excision repair(NER), focusing on post-translational modifications(ubiquitylation and sumoylation) of the damage recognition proteins, XPC and DDB2. Concerning the UV-induced reversible ubiquitylation of XPC, knockdown of the identified deubiquitylation factor was found to result in accumulation of ubiquitylated XPC, which significantly compromised recruitment of XPC to damaged DNA sites. We also succeeded in reconstitution of in vitro sumoylation of XPC and identification of the modification sites. The cells stably expressing the non-sumoylated mutant XPC exhibited retardation of NER and impaired induction of XPC ubiquitylation after UV irradiation, possibly because the functional interaction between XPC and DDB2 may be mediated by SUMO. Finally, target sites for DDB2 self-ubiquitylation by the CRL4^<DDB2> ubiquitin ligase were identified, suggesting the possibility that DDB2 ubiquitylation may have distinct roles, protein degradation and dissociation from damaged DNA, depending on the modification sites.

Research Products

• [Journal Article] Multiple DNA damage recognition factors involved in mammalian nucleotide excision repair (2011)
  • Author(s): Sugasawa, K
  • Journal Title: Biochemistry(Moscow) Volume: 76 Pages: 16-23
  • Peer Reviewed
• [Journal Article] The xeroderma pigmentosum pathway : decision tree analysis of DNA quality (2011)
  • Author(s): Naegeli, H. and Sugasawa, K.
  • Journal Title: DNA Repair Volume: 10 Pages: 673-683
  • Peer Reviewed
• [Journal Article] NBS1 recruits RAD18 via a RAD6-like domain and regulates Pol h-dependent translesion DNA synthesis (2011)
  • Author(s): Yanagihara, H., Kobayashi, J., Tateishi, S., Kato, A., Matsuura, S., Tauchi, H., Yamada, K., Takezawa, J., Sugasawa, K., Masutani, C., Hanaoka, F., Weemaes, C. M., Mori, T., Zou, L., and Komatsu, K.
  • Journal Title: Mol. Cell Volume: 43 Pages: 788-797
  • Peer Reviewed
• [Journal Article] The molecular basis of CRL4^<DDB2/CSA> ubiquitin ligase architecture (2011)
  • Author(s): Fischer, E. S., Scrima, A., Bohm, K., Matsumoto, S., Lingaraju, G. M., Faty, M., Yasuda, T., Cavadini, S., Wakasugi, M., Hanaoka, F., Iwai, S., Gut, H., Sugasawa, K., and Thoma, N. H.
  • Journal Title: targeting, and activation. Cell Volume: 147 Pages: 1024-1039
  • Peer Reviewed
• [Journal Article] DNA damage recognition for mammalian global genome nucleotide excision repair (2011)
  • Author(s): Sugasawa, K.
  • Journal Title: DNA Repair Pages: 453-476
  • Peer Reviewed
• [Journal Article] Fluorescent probes for the analysis of DNA strand scission in base excision repair (2010)
  • Author(s): Matsumoto, N., Toga, T., Hayashi, R., Sugasawa, K., Katayanagi, K., Ide, H., Kuraoka, I., and Iwai, S.
  • Journal Title: Nucleic Acids Res Volume: 38 Pages: e101
  • Peer Reviewed
• [Journal Article] Stimulation of DNA glycosylase activities by XPC protein complex : roles of protein-protein interactions (2010)
  • Author(s): Shimizu, Y., Uchimura, Y., Dohmae, N., Saitoh, H., Hanaoka, F., and Sugasawa, K.
  • Journal Title: J. Nucleic Acids 2010 Pages: 805698
  • Peer Reviewed
• [Journal Article] An EF-hands protein centrin-1 is an EGTA-sensitive SUMO-interacting protein in mouse testis (2010)
  • Author(s): Tanaka, N., Goto, M., Kawasaki, A., Sasano, T., Eto, K., Nishi, R., Sugasawa, K., Abe, S., and Saitoh, H.
  • Journal Title: Cell Biochem. Funct Volume: 28 Pages: 604-612
  • Peer Reviewed
• [Journal Article] UV-DDB-dependent regulation of nucleotide excision repair kinetics in living cells (2009)
  • Author(s): Nishi, R., Alekseev, S., Dinant, C., Hoogstraten, D., Houtsmuller, B. A., Hoeijmakers, J. H. J., Vermeulen, W., Hanaoka, F., and Sugasawa, K.
  • Journal Title: DNA Repair Volume: 8 Pages: 767-776
  • Peer Reviewed
• [Journal Article] 色素性乾皮症遺伝子産物による (2009)
  • Author(s): 菅澤薫
  • Journal Title: DNA損傷認識機構医学のあゆみ Volume: 228 Pages: 137-142
• [Journal Article] UV-DDB : A molecular machine linking DNA repair with ubiquitination (2009)
  • Author(s): Sugasawa, K.
  • Journal Title: DNA Repair Volume: 8 Pages: 969-972
  • Peer Reviewed
• [Journal Article] Regulation of damage recognition in mammalian global genomic nucleotide excision repair (2009)
  • Author(s): Sugasawa, K.
  • Journal Title: Mutat. Res Volume: 685 Pages: 19-37
  • Peer Reviewed
• [Journal Article] The CUL4 enigma : culling DNA repair factors (2009)
  • Author(s): Sugasawa, K.
  • Journal Title: Mol. Cell Volume: 34 Pages: 403-404
• [Journal Article] Two-step recognition of DNA damage for mammalian nucleotide excision repair : directional binding of the XPC complex and DNA strand scanning (2009)
  • Author(s): Sugasawa, K., Akagi, J., Nishi, R., Iwai, S., and Hanaoka, F.
  • Journal Title: Mol. Cell Volume: 36 Pages: 642-653
  • Peer Reviewed
• [Journal Article] Fluorescence correlation spectroscopy of the binding of nucleotide excision repair protein XPChHr23B with DNA substrates (2008)
  • Author(s): Roche, Y., Zhang, D., Segers-Nolten, G. M., Vermeulen, W., Wyman, C., Sugasawa, K., Hoeijmakers, J., and Otto, C.
  • Journal Title: J. Fluoresc Volume: 18 Pages: 987-995
  • Peer Reviewed
• [Journal Article] XPC, its product and biological roles (2008)
  • Author(s): Sugasawa, K.
  • Journal Title: Adv. Exp. Med. Biol Volume: 637 Pages: 47-56
  • Peer Reviewed
• [Presentation] Roles for sumoylation in regulation of the xeroderma pigmentosum group C protein (2011)
  • Author(s): Akita, M., Tak, Y.-S., Shimura, T., Hanaoka, F., and Sugasawa, K.
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20111213-16
• [Presentation] Functional analysis of the CRL4^<DDB2> E3 ligase in mammalian nucleotide excision repair (2011)
  • Author(s): Matsumoto, S., Yasuda, T., Dohmae, N., Fischer, E. S., Thoma, N. H., Nishi, R., Hanaoka, F., and Sugasawa, K.
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20111213-16
• [Presentation] Reconstitution of mammalian nucleotide excision repair : molecular basis for DNA damage recognition (2011)
  • Author(s): Sugasawa, K., Tone, D., Yasuda, T., and Hanaoka, F.
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20111213-16
• [Presentation] DNA damage recognition in mammalian nucleotide excision repair : molecular mechanism and involvement of chromatin structure (2011)
  • Author(s): Sugasawa, K.
  • Organizer: The 27th RBC-NIRS International Symposium on Chromatin Dynamics and Epigenetic Memory
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 20111209-10
• [Presentation] ヌクレオチド除去修復におけるDNA損傷認識の分子基盤 (2011)
  • Author(s): 菅澤薫
  • Organizer: 日本放射線影響学会第54回大会
  • Place of Presentation: 神戸
  • Year and Date: 20111117-19
• [Presentation] Regulation of DNA damage recognition in mammalian nucleotide excision repair (2011)
  • Author(s): Sugasawa, K.
  • Organizer: Conference on Responses to DNA damage : from molecular mechanism to human disease
  • Place of Presentation: The Netherlands
  • Year and Date: 20110403-08
• [Presentation] A 19S proteasome subunit PSMD14 is involved in deubiquitination of XPC (2010)
  • Author(s): Akagi, J., Hanaoka, F., and Sugasawa, K.
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会(BMB2010)
  • Place of Presentation: 神戸
  • Year and Date: 20101207-10
• [Presentation] Functional analysis of the N-terminal domain of the DNA damage recognition protein DDB2 (2010)
  • Author(s): Matsumoto, S., Yasuda, T., Dohmae, N., Murakami, K., Nishi, R., Hanaoka, F., and Sugasawa, K.
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会(BMB2010)
  • Place of Presentation: 神戸
  • Year and Date: 20101207-10
• [Presentation] Roles for centrin-2 in DNA damage recognition for global genome nucleotide excision repair (2010)
  • Author(s): Nishi, R., Tak, Y.-S., Sakai, W., Hanaoka, F., and Sugasawa, K.
  • Organizer: The 7th International Symposium on DNA Replication, Recombination and Repair(3R Symposium)
  • Place of Presentation: Toyama, Japan
  • Year and Date: 20101026-30
• [Presentation] Molecular basis for DNA damage recognition by xeroderma pigmentosum gene products (2010)
  • Author(s): 菅澤薫、花岡文雄
  • Organizer: 第69回日本癌学会学術総会
  • Place of Presentation: 大阪
  • Year and Date: 20100922-24
• [Presentation] ゲノム全体を監視するDNA損傷認識の分子基盤 (2010)
  • Author(s): 菅澤薫、安田武嗣、赤木純一、西良太郎、岩井成憲、花岡文雄
  • Organizer: 第9回核ダイナミクス研究会
  • Place of Presentation: 伊豆
  • Year and Date: 20100527-29
• [Presentation] DNA damage recognition mechanism for mammalian nucleotide excision repair (2010)
  • Author(s): Sugasawa, K.
  • Organizer: International Conference on Radiation and Cancer Biology
  • Place of Presentation: Nagasaki, Japan
  • Year and Date: 20100217-20
• [Presentation] Post-translational modification of XPC protein controls global genome nucleotide excision repair (2009)
  • Author(s): Tak, Y.-S., Shimura, T., Nishi, R., Okuda-Shimizu, Y., Shimizu, Y., Hanaoka, F., Sugasawa, K.
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20091209-12
• [Presentation] DNA damage recognition coordinated by the multiple xeroderma pigmentosum gene products (2009)
  • Author(s): 菅澤薫
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 20091209-12
• [Presentation] Post-translational modifications involved in cellular DNA damage response (2009)
  • Author(s): Sugasawa, K.
  • Organizer: 2009 Japan-Taiwan Joint Symposium on Cell Signaling and Gene Regulation
  • Place of Presentation: Kobe, Japan
  • Year and Date: 20091111-12
• [Presentation] Coordinated actions of multiple DNA damage detectors in nucleotide excision repair (2009)
  • Author(s): Sugasawa, K.
  • Organizer: National Conference on Medical Genomics and Proteomics
  • Place of Presentation: Novosibirsk, Russia
  • Year and Date: 20090909-10
• [Presentation] Post-translational modification of XPC protein controls global genome nucleotide excision repair (2009)
  • Author(s): Tak, Y.-S., Shimura, T., Nishi, R., Okuda-Shimizu, Y., Shimizu, Y., Hanaoka, F., and Sugasawa, K.
  • Organizer: The 3rd ASM Conference on DNA Repair and Mutagenesis
  • Place of Presentation: Whistler, Canada
  • Year and Date: 20090530-0605
• [Presentation] Molecular basis for DNA damage recognition in nucleotide excision repair (2009)
  • Author(s): Sugasawa, K.
  • Organizer: International Symposium on DNA Damage Response and Repair Mechanisms
  • Place of Presentation: Crete, Greece
  • Year and Date: 20090420-23
• [Presentation] ヌクレオチド除去修復におけるTFIIHへリカーゼ活性調節機構の解析 (2008)
  • Author(s): 赤木純一、菅澤薫
  • Organizer: 第31回日本分子生物学会年会・第81回日本生化学会大会(BMB2008)
  • Place of Presentation: 神戸
  • Year and Date: 20081209-12
• [Presentation] Centrin 2 enhances in vitro nucleotide excision repair via complex formation with xeroderma pigmentosum group C pretein through its C-terminal domain (2008)
  • Author(s): Nishi, R., Miyake, H., Murakami, K., Kitano, K., Hanaoka, F., and Sugasawa, K.
  • Organizer: 第31回日本分子生物学会年会・第81回日本生化学会大会(BMB2008)
  • Place of Presentation: 神戸
  • Year and Date: 20081209-12
• [Presentation] ヌクレオチド除去修復におけるDNA損傷認識の分子機序 (2008)
  • Author(s): 菅澤薫
  • Organizer: 第31回日本分子生物学会年会・第81回日本生化学会大会(BMB2008)
  • Place of Presentation: 神戸
  • Year and Date: 20081209-12
• [Presentation] Xeroderma pigmentosum proteins and their functions in DNA damage recognition (2008)
  • Author(s): 菅澤薫
  • Organizer: 第67回日本癌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 20081028-30
• [Presentation] Analyses of regulatory mechanisms of TFIIH helicase activity in nucleotide exicion repair (2008)
  • Author(s): Akagi, J. and Sugasawa, K.
  • Organizer: The 6th International Symposium on DNA Replication, Recombination and Repair(3R Symposium)
  • Place of Presentation: Kakegawa, Shizuoka
  • Year and Date: 20081027-30
• [Presentation] Molecular mechanisms underlying efficient DNA damage recognition for nucleotide excision repair (2008)
  • Author(s): Sugasawa, K., Nishi, R., Akagi, J., Tak, Y.-S., Kobayashi, H., and Hanaoka, F.
  • Organizer: The 6th International Symposium on DNA Replication, Recombination and Repair(3R Symposium)
  • Place of Presentation: Kakegawa, Shizuoka
  • Year and Date: 20081027-30
• [Presentation] Damage recognition mechanism for mammalian nucleotide excision repair (2008)
  • Author(s): Sugasawa, K.
  • Organizer: Russian-European Workshop on DNA Repair and Epigenetic Regulation of Genome Stability
  • Place of Presentation: St. Petersburg, Russia
  • Year and Date: 20080624-26
• [Remarks] ホームページ
  • URL: http://www.research.kobe-u.ac.jp/brce-sugasaw