Molecular mechanisms of translesion DNA synthesis and its involvement in carcinogenesis

2009 Fiscal Year Final Research Report

• Project/Area Number: 17013053
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Gakushuin University (2008-2009); Osaka University (2005-2007)
• Principal Investigator: HANAOKA Fumio Gakushuin University, 理学部, 教授 (50012670)
• Co-Investigator(Kenkyū-buntansha): YOKOI Masayuki 大阪大学, 生命機能研究科, 助教 (00322701) ; IWAI Shigenori 大阪大学, 基礎工学研究科, 教授 (10168544)
• Co-Investigator(Renkei-kenkyūsha): YOKOI Masayuki 学習院大学, 理学部, 助教 (00322701)
• Project Period (FY): 2005 – 2009
• Keywords: がん / 遺伝子 / ゲノム安定性 / DNAポリメラーゼ / DNA修復 / 損傷乗り越え複製 / ノックアウトマウス / 紫外線

Research Abstract

We have clarified the mechanism in which human DNA polymerase η (Polη) interacts with other proteins for switching between Polη and the replicative polymerase at stalled replication forks. We have solved high-resolution crystal structures of human Polη at four consecutive steps during DNA synthesis through cis-syn cyclobutane thymine dimers. In addition, we have generated Polηknockout mice and observed high incidence of skin tumors upon UV-B irradiation in these mice.

Research Products

• [Journal Article] Simultaneous disruption of two DNA polymerases, Polη and Polζ, in Avian DT40 cells unmasks the role of Polη in cellular response to various DNA lesions. (2010)
  • Author(s): Hirota, K., Sonoda, E., Kawamoto, T., Motegi, A., Masutani, C., Hanaoka, F., Szuts, D., Iwai, S., Sale, J.E., Lehmann, A., Takeda, S.
  • Journal Title: PLoS.Genet. 6 Pages: ii:e1001151
  • Peer Reviewed
• [Journal Article] Structure and mechanism of human DNA polymeraseη. (2010)
  • Author(s): Biertampfel, C., Zhao, Y., Kondo, Y., Ramon-Maiques, S., Gregory, M., Lee, J.Y., Masutani, C., Lehmann, A.R., Hanaoka, F., Yang, W.
  • Journal Title: Nature 465 Pages: 1044-1049
  • Peer Reviewed
• [Journal Article] The molecular chaperone Hsp90 regulates accumulation of DNA polymerase eta at replication stalling sites in UV-irradiated cells. (2010)
  • Author(s): Sekimoto, T., Oda, T., Pozo, F.M., Murakumo, Y., Masutani, C., Hanaoka, F., Yamaashita, T.
  • Journal Title: Hal.Cell 37 Pages: 79-89
  • Peer Reviewed
• [Journal Article] The chromosomal association of condensin II is required by a noncatalytic function of PP2A. (2009)
  • Author(s): Takemoto, A., Maeshima, K., Ikehara, T., Yamaguchi, K., Murayama, A., Imamura, S., lamamoto, N., Yokoyama, S., Hirano, T., Watanabe, Y., Hanaoka, F., Yanagisawa, J., Kimura, K.
  • Journal Title: Nat.Struct.Aiol.Biol. 16 Pages: 1302-1308
  • Peer Reviewed
• [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., Hanaoka, F.
  • Journal Title: Alai.Cell 36 Pages: 642-653
  • Peer Reviewed
• [Journal Article] Aberrant DNA polymerase alpha is excluded from the nucleus by defective import and degradation in the nucleus. (2009)
  • Author(s): Eichinger, C.S., Mizuno, T., Mizuno, K., Miyake, Y., Yanagi, K., Imamoto, N., Hanaoka, F.
  • Journal Title: J.Biol.Chem. 284 Pages: 30604-30614
  • Peer Reviewed
• [Journal Article] Interaction with DNA polymerase eta is required for nuclear accumulation of REV1 and suppression of spontaneous mutations in human cells. (2009)
  • Author(s): Akagi, J., Masutani, C., Kataoka, Y., Kan, T., Ohashi, E., Mori, T., Ohmori, H., Hanaoka, F.
  • Journal Title: DM Repair 8 Pages: 585-599
  • Peer Reviewed
• [Journal Article] Cyclin A-Cdkl regulates the origin firing program in mammalian cells. (2009)
  • Author(s): Katsuno, Y., Suzuki, A., Sugimura, K., Okumura, K., Zineldeen, D.H., Shimada, M., Niida, H., Mizuno, T., Hanaoka, F., Nakanishi, M.
  • Journal Title: Proc.Natl.Acad Sci.USA 106 Pages: 3184-3189
  • Peer Reviewed
• [Journal Article] Schizosaccharomyees pombe Ddbl recruits substrate-specific adaptor proteins through a novel protein motif, the DDB-box. (2008)
  • Author(s): Fukumoto, Y., Dohmae, N., Hanaoka, F.
  • Journal Title: Mol.Cell.Biol. 28 Pages: 6746-6756
  • Peer Reviewed
• [Journal Article] In vivo destabilization and functional defects of the xeroderma pigmentosum C protein caused by a pathogenic missense mutation. (2007)
  • Author(s): Yasuda, G., Nishi, R., Watanabe, E., Mori, T., Iwai, S., Orioli, D., Stefanini, M., Hanaoka, F., Sugasawa, K.
  • Journal Title: Mol.Cell.Biol. 27 Pages: 6606-6614
  • Peer Reviewed
• [Journal Article] A genome-wide survey and systematic RNAi-based characterization of helicase-like genes in Caenorhabditis elegans. (2007)
  • Author(s): Eki, T., Ishihara, T., Katsura, I., Hanaoka, F.
  • Journal Title: DNA Res. 14 Pages: 183-199
  • Peer Reviewed
• [Journal Article] Rad54 dissociates homologous recombination intermediates by branch migration. (2007)
  • Author(s): Bugreev, D.V., Hanaoka, F., Mazin, A.V.
  • Journal Title: Nat.Struct.Mol.Biol. 14 Pages: 746-753
  • Peer Reviewed
• [Journal Article] Structure of the human GINS complex and its assembly and functional interface in replication initiation. (2007)
  • Author(s): Kamada, K., Kubota, Y., Arata, T., Shindo, Y., Hanaoka, F.
  • Journal Title: Nature Struct.Mot.Biol. 14 Pages: 388-396
  • Peer Reviewed
• [Journal Article] Negative regulation of condensin I by CK2-mediated phosphorylation. (2006)
  • Author(s): Takemoto, A., Kimura, K., Yanagisawa, J., Yokoyama, S., Hanaoka, F.
  • Journal Title: EMBO J. 25 Pages: 5339-5348
  • Peer Reviewed
• [Journal Article] Ultraviolet B radiation induces epithelial tumors in mice lacking pol η and mesenchymal tumors in mice deficient for pol (2006)
  • Author(s): Ohkumo, T., Kondo, Y., Yokoi, M., Tsukamoto, T., Yamada, A., Sugimoto, T., Kanao, R., Higashi, Y., Kondoh, H., Tatematsu, M., Masutani, C., Hanaoka, F.
  • Journal Title: ι.Mal.Cell.Biol. 26 Pages: 7696-7706
  • Peer Reviewed
• [Journal Article] A human DNA polymerase eta complex containing Rad18, Rad6 and Revl ; proteomic analysis and targeting of the complex to the chromatin-bound fraction of cells undergoing replication fork arrest. (2006)
  • Author(s): Yuasa, M.S., Masutani, C., Hirano, A., Cohn, M.A., Yamaizumi, M., Nakatani, Y., Hanaoka, F.
  • Journal Title: Genes Cells 11 Pages: 731-744
  • Peer Reviewed
• [Journal Article] Conserved XPB core structure and motifs for DNA unwinding: implications for pathway selection of transcription or excision repair. (2006)
  • Author(s): Fan, L., Arvai, A.S., Cooper, P.K., Iwai, S., Hanaoka, F., Tainer, J.A.
  • Journal Title: Mot.Cell 22 Pages: 27-37
  • Peer Reviewed
• [Journal Article] Deficiency of the Caenorhabditis elegans DNA polymerase eta homologue increases sensitivity to UV radiation during germ-line development. (2006)
  • Author(s): Ohkumo, T., Masutani, C., Eki, T., Hanaoka, F.
  • Journal Title: Cell Struct.Funct. 31 Pages: 29-37
  • Peer Reviewed
• [Journal Article] Alterations of DNA and chromatin structures at telomeres and genetic instability in mouse cells defective in DNA polymerase α. (2005)
  • Author(s): Nakamura, M., Nabetani, A., Mizuno, T., Hanaoka, F., Ishikawa, F.
  • Journal Title: Mal.Cell.Biol. 25 Pages: 11073-11088
  • Peer Reviewed
• [Journal Article] Azathioprine and UVA light generate mutagenic oxidative DNA damage. (2005)
  • Author(s): O'Donovan, P., Perrett, C.M., Zhang, X., Montaner, B., Xu, Y.Z., Harwood, C.A., McGregor, J.M., Walker, S.L., Hanaoka, F., Karran, P.
  • Journal Title: Science 309 Pages: 1871-1874
  • Peer Reviewed
• [Journal Article] Conformational change in the catalytic site of the ribonuclease YoeB toxin by YefM antitoxin. (2005)
  • Author(s): Kamada, K., Hanaoka, F.
  • Journal Title: Mal.Cell 19 Pages: 497-509
  • Peer Reviewed
• [Journal Article] Centrin 2 stimulates nucleotide excision repair by interacting with xeroderma pigmentosum group C protein. (2005)
  • Author(s): Nishi, R., Okuda, Y., Watanabe, E., Mori, T., Iwai, S., Masutani, C., Sugasawa, K., Hanaoka, F.
  • Journal Title: Cell.Biol. 25 Pages: 5664-5674
  • Peer Reviewed
• [Journal Article] Crystal structure of SUMO1-conjugated thymine DNA glycosylase. (2005)
  • Author(s): Baba, D., Maita, N., Jee, J.-G., Uchimura, Y., Saitoh, H., Sugasawa, K., Hanaoka, F., Tochio, H., Hiroaki, H., Shirakawa, M.
  • Journal Title: Nature 435 Pages: 979-982
  • Peer Reviewed
• [Journal Article] Different mutation signatures in DNA polymerase η- and MSH6-deficient mice suggest separate roles in antibody diversification. (2005)
  • Author(s): Martomo, S.A., Yang, W.W., Wersto, R.P., Ohkumo, T., Kondo, Y., Yokoi, M., Masutani, C., Hanaoka, F., Gearhart, P.J.
  • Journal Title: Proc.Natl.Acad Sci.USA 102 Pages: 8656-8661
  • Peer Reviewed
• [Journal Article] UV-induced ubiquitylation of XPC protein mediated by UV-DDB-ubiquit in ligase complex. (2005)
  • Author(s): Sugasawa, K., Okuda, Y., Sai jo, M., Nishi, R., Matsuda, N., Chu, G., Mori, T., Iwai, S., Tanaka, K., Tanaka, K., Hanaoka, F.
  • Journal Title: Cell 121 Pages: 387-400
  • Peer Reviewed
• [Journal Article] Nucleosomal structure of undamaged DNA regions suppresses the non-specific DNA binding of the XPC complex. (2005)
  • Author(s): Yasuda, T., Sugasawa, K., Shimizu, Y., Iwai, S., Shiomi, T., Hanaoka, F.
  • Journal Title: DNA Repair 4 Pages: 389-395
  • Peer Reviewed
• [Presentation] Regulation of translesion DNA synthesis with special emphasis on DNA polymerase η and PCNA ubiquitylation. (2010)
  • Author(s): Hanaoka, F., Deguchi, S., Kanao, R., Masutani, C.
  • Organizer: EMBO Workshop, The Interface between the Ubiquitin Family and the DNA Damage Response
  • Place of Presentation: Rovinj, Croatia
  • Year and Date: 20100901-20100905
• [Presentation] Tumorigenesis induced by chronic treatment with UV-B in Pol η- and Xpc-deficient mice. (2010)
  • Author(s): Hanaoka, F.
  • Organizer: Gordon Research Conference, Mutagenesis
  • Place of Presentation: Waterville, USA
  • Year and Date: 20100801-20100806
• [Presentation] UV-induced mutations in the epidermal genome of TLS polymerase-deficient mice. (2009)
  • Author(s): Hanaoka, F.
  • Organizer: The 40^<th> International Symposium of the Princess Takamatsu Cancer Research Fund
  • Place of Presentation: 東京
  • Year and Date: 20091110-20091112
• [Presentation] UV-induced mutations in the epidermal genome of DNA polymerase η- and ι-deficient mice occurring in vivo. (2009)
  • Author(s): Hanaoka, F.
  • Organizer: Second Erling Seeberg Symposium on DNA Repair, Keynote Address
  • Place of Presentation: Alesund and Geiranger, Nnrwav
  • Year and Date: 20090620-20090625
• [Presentation] ゲノム情報維持の分子機構に関する研究 (2009)
  • Author(s): 花岡文雄
  • Organizer: 日本薬学会第129年会日本薬学会賞受賞講演
  • Place of Presentation: 京都
  • Year and Date: 20090326-20090328
• [Presentation] 損傷を乗り越えるポリメラーゼの機能と発癌の制御東北大学・加齢医学研究所・シンポジウム (2009)
  • Author(s): 花岡文雄
  • Organizer: 平成20年度・ゲノムリサーチセンター・ワークショップ「癌と老化を乗り越えるDNA修復の科学」
  • Place of Presentation: 仙台
  • Year and Date: 2009-03-13
• [Presentation] 損傷乗り越え型DNAポリメラーゼ・イータおよびイオタ欠損マウスの皮膚における紫外線誘発発がんと突然変異 (2008)
  • Author(s): 花岡文雄
  • Organizer: 日本放射線影響学会第51回大会、シンポジウム「放射線発がんの基盤を形成する遺伝子・細胞・組織の応答」
  • Place of Presentation: 北九州
  • Year and Date: 20081119-20081121
• [Presentation] Schizosaccharomyces pombe Ddbl recruits substrate-specific adaptor proteins through a novel protein motif, the DDB-box. (2008)
  • Author(s): Fukumoto, Y., Dohmae, N., Hanaoka, F.
  • Organizer: The 6th 3R Symposium (3R Symposium 2008)
  • Place of Presentation: 掛川
  • Year and Date: 20081027-20081030
• [Presentation] In vivo and in vitro studies on DNA polymerase eta. (2008)
  • Author(s): Hanaoka, F.
  • Organizer: Gordon Research Conference on Mutagenesis
  • Place of Presentation: Oxford, UK
  • Year and Date: 20080720-20080725
• [Presentation] 損傷乗り越え複製研究の最前線 (2008)
  • Author(s): 花岡文雄
  • Organizer: 東京都臨床医学総合研究所
  • Place of Presentation: 東京
  • Year and Date: 2008-12-19
• [Presentation] 損傷乗り越え複製とがん (2008)
  • Author(s): 花岡文雄
  • Organizer: 第67回日本癌学会学術総会、モーニング・レクチャー
  • Place of Presentation: 名古屋
  • Year and Date: 2008-10-29
• [Presentation] Translesion synthesis and cancer. (2008)
  • Author(s): Hanaoka, F.
  • Organizer: Clare Hall Laboratories, Cancer Research UK
  • Place of Presentation: South Mimms, UK
  • Year and Date: 2008-07-25
• [Presentation] 損傷乗り越え複製の分子機構-DNAポリメラーゼ・イータを中心に- (2007)
  • Author(s): 花岡文雄、横井雅幸、大雲剛志、金尾梨絵、赤木純一、益谷央豪
  • Organizer: 第30回日本分子生物学会年会・第80回日本生化学会大会合同大会(BMB2007)、シンポジウム「DNA修復研究の最前線」
  • Place of Presentation: 横浜
  • Year and Date: 20071211-20071215
• [Presentation] 損傷轍複製とがん (2007)
  • Author(s): 花岡文雄、益谷央豪、塚本徹哉、立松正衛
  • Organizer: 第66回日本癌学会学術総会、シンポジウム「ゲノム不安定性」
  • Place of Presentation: 横浜
  • Year and Date: 20071003-20071005
• [Presentation] TLS polymerase deficiencies and cancer. (2007)
  • Author(s): Hanaoka, F.
  • Organizer: The 3rd Japan-US DNA Repair Meeting
  • Place of Presentation: 仙台
  • Year and Date: 20070507-20070511
• [Presentation] Translesion synthesis and cancer. (2007)
  • Author(s): Hanaoka, F.
  • Organizer: 7th Joint Conference of the American Association for Cancer Research and the Jananese Cancer Association.
  • Place of Presentation: Waikoloa, Hawaii, USA
  • Year and Date: 20070421-20070425
• [Presentation] Recent studies on mammalian DNA polymerase eta (2007)
  • Author(s): Hanaoka, F.
  • Organizer: the product of XP variant responsible gene. Mayo Clinic
  • Place of Presentation: Rochester, Minnesota, USA
  • Year and Date: 2007-05-21
• [Presentation] Defective translesion DNA synthesis and links to human disease. (2007)
  • Author(s): Hanaoka, F.
  • Organizer: 9th Annual Midwest DNA Repair Symposium, Keynote Lecture
  • Place of Presentation: Columbus, Ohio
  • Year and Date: 2007-05-20
• [Presentation] DNA repair and ubiquitin. (2007)
  • Author(s): Hanaoka, F.
  • Organizer: Ohio State University Medical Center
  • Place of Presentation: Columbus, Ohio, USA
  • Year and Date: 2007-05-18
• [Presentation] DNA損傷乗り越え複製と発がん (2007)
  • Author(s): 花岡文雄
  • Organizer: がん特定研究否同シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2007-02-23
• [Presentation] In vivo and in vitro studies on DNA polymerase eta, the product of XP variant gene. (2006)
  • Author(s): Hanaoka, F.
  • Organizer: The First International Symposium on the MEXT Priority Research Project.-The Chromosome Cycle-.
  • Place of Presentation: 東京
  • Year and Date: 20060626-20060627
• [Presentation] In vivo and in vitro studies on mammalian DNA polymerase eta, the product of XP-V responsible gene. (2006)
  • Author(s): Hanaoka, F.
  • Organizer: Erling Seeberg Symposium on DNA Repair.
  • Place of Presentation: Bodo and Henningsver, Lofoten, Norway
  • Year and Date: 20060528-20060602
• [Presentation] 遺伝盾報を保持するしくみ (2006)
  • Author(s): 花岡文雄
  • Organizer: 第16回遺伝医学セミナー
  • Place of Presentation: 大阪
  • Year and Date: 2006-09-02
• [Presentation] 色素性乾皮症バリアント群の解析から明らかにった新規DNA損傷応答について (2006)
  • Author(s): 花岡文雄
  • Organizer: 山泉克教授追悼講演会「DNA傷害と修復応答」
  • Place of Presentation: 熊本
  • Year and Date: 2006-07-21
• [Presentation] 紫外線障害修復機構-その概要とXPV遺伝子欠損ラウスを用いた皮膚発がん実験について- (2006)
  • Author(s): 花岡文雄
  • Organizer: 第28回日本光医学・光生物学会
  • Place of Presentation: 徳島
  • Year and Date: 2006-07-07
• [Presentation] DNA修復におけるユビキチン化の役割 (2006)
  • Author(s): 花岡文雄
  • Organizer: 千里ライフサイエンスセミナー「蛋白質の修飾と機能制御」
  • Place of Presentation: 大阪
  • Year and Date: 2006-07-05
• [Remarks] ホームページ等
  • URL: http://www.gakushuin.ac.jp/univ/sci/bio/laboratory/detail_hanaoka/theme.html
• [Patent(Industrial Property Rights)] DNA損傷修復剤とスクリーニング方法 (2009)
  • Inventor(s): 菅澤薫、花岡文雄、清水祐一郎
  • Industrial Property Rights Holder: 科学技術振興機構、(独)理化学研究所
  • Industrial Property Number: 特許,特許第4402349号
  • Acquisition Date: 2009-11-06
• [Patent(Industrial Property Rights)] 新規ヒトトポイソメラーゼ2α阻害蛋白質及びその利用 (2005)
  • Inventor(s): 中西啓、花岡文雄、大海忍
  • Industrial Property Rights Holder: (独)理化学研究所
  • Industrial Property Number: 特許,特許第3665775号
  • Acquisition Date: 2005-04-08