Analyses of functional roles of TLS polymerases using transgenic mice

• Project/Area Number: 22249005
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Gakushuin University
• Principal Investigator: HANAOKA Fumio 学習院大学, 理学部, 教授 (50012670)
• Co-Investigator(Renkei-kenkyūsha): YOKOI Masayuki 学習院大学, 理学部, 助教 (00322701) ; AKAGI Jun-ichi 国立医薬品食品衛生研究所, 病理部, 研究員 (60512556)
• Project Period (FY): 2010-04-01 – 2013-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥48,750,000 (Direct Cost: ¥37,500,000、Indirect Cost: ¥11,250,000); Fiscal Year 2012: ¥13,000,000 (Direct Cost: ¥10,000,000、Indirect Cost: ¥3,000,000); Fiscal Year 2011: ¥13,000,000 (Direct Cost: ¥10,000,000、Indirect Cost: ¥3,000,000); Fiscal Year 2010: ¥22,750,000 (Direct Cost: ¥17,500,000、Indirect Cost: ¥5,250,000)
• Keywords: 遺伝子ノックアウト / 損傷乗り越え複製 / 紫外線 / DNAポリメラーゼ / 突然変異 / 損傷トレランス / TLSポリメラーゼ / DNAポリメラーゼκ / REV1 / ノックアウトマウス / (6-4)光産物 / DNA損傷トレランス / DNAポリメラーゼ・イータ / 紫外線感受性

Research Abstract

In order to examine the functional roles of translesion synthesis (TLS) in mammalian cells, we have established many mouse embryonic fibroblast cell lines expressing polymerase activity-defective or Rev1 interaction-defective mutant DNA polymerase eta (Pol eta) in Pol eta, Pol iota or Pol kappa single-, double- or triple-deficient mouse embryonic fibroblasts (MEFs). Using these MEF cells, we investigated UV sensitivity as well as mutation frequency upon UV irradiation. Our results suggest that there is an error-prone TLS pathway that uses Pol kappa via Rev1 in the absence of Pol eta.

Research Products

• [Journal Article] A Cyclobutane thymine-N4- methylcytosine dimer is resistant to hydrolysis but strongly blocks DNA synthesis (2014)
  • Author(s): Yamamoto J, Oyama T, Kunishi T, Masutani C, Hanaoka F, Iwai S
  • Journal Title: Nucleic Acids Res. Volume: 42 Pages: 2075-2084
  • DOI: 10.1093/nar/gkt1039
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [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] Mechanism of somatic hyper- mutation at the WA motif by human DNA polymeraseη (2013)
  • Author(s): Zhao Y, Gregory MT, Biertümpfel C, Hua YJ, Hanaoka F, Yang W
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 110 Pages: 8146-8151
  • DOI: 10.1073/pnas.1303126110
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Trapping DNA replication origins from the human genome (2013)
  • Author(s): Eki, T., Murakami, Y., and Hanaoka, F
  • Journal Title: Genes (Basel) Volume: 4 Pages: 198-225
  • Peer Reviewed
• [Journal Article] Trapping DNA replication origins from the human genome (2013)
  • Author(s): Eki T, Murakami Y, Hanaoka F
  • Journal Title: Genes(Basel) Volume: 4 Pages: 198-225
  • DOI: 10.3390/genes4020198
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification of new scavengers for hydroxyl radicals and superoxide dismutase by utilizing ultraviolet A photoreaction of 8-methoxypsoralen and a variety of mutants of Escherichia coli : implications on certain diseases of DNA repair deficiency (2012)
  • Author(s): Ahmad, S., Yokoi, M., and Hanaoka, F
  • Journal Title: J. Photochem. Photobiol. B. Volume: 116 Pages: 30-36
  • Peer Reviewed
• [Journal Article] A DNA oligomer containing 2,2,4-triamino-5(2H)- oxazolone is incised by human NEIL1 and NTH1 (2012)
  • Author(s): Kino K
  • Journal Title: Mut. Res. Volume: 734 Pages: 73-77
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• [Journal Article] Structural basis of human DNA polymeraseη-mediated chemoresistance to cisplatin (2012)
  • Author(s): Zhao Y
  • Journal Title: Proc. Natl. Acad. Sci. Volume: 109 Pages: 7269-7274
  • DOI: 10.1073/pnas.1202681109
  • Peer Reviewed
• [Journal Article] The vital role of polζand REV1 in mutagenic, but not correct, DNA synthesis across benzo[a]pyrene-dG and the recruitment of polζby REV1 to a replication-stalled site (2012)
  • Author(s): Hashimoto, K., Cho, Y., Yang, I. Y., Akagi, J., Ohashi, E., Tateishi, S., de Wind, N., Hanaoka, F., Ohmori, H., and Moriya, M
  • Journal Title: J. Biol. Chem. Volume: 96 Pages: 9613-9622
  • Peer Reviewed
• [Journal Article] Analysis of mice deficient in both REV1 catalytic activity and POLH reveals an unexpected role for POLH in the generation of C to G and G to C transversion during Ig gene hypermutation (2012)
  • Author(s): Kano C
  • Journal Title: Int. Immunol. Volume: 24 Pages: 169-174
  • DOI: 10.1093/intimm/dxr109
  • Peer Reviewed
• [Journal Article] Stalled Polηat its cognate substrate initiates an alternative translesion synthesis pathway via interaction with REV1 (2012)
  • Author(s): Ito W
  • Journal Title: Genes Cells Volume: 17 Pages: 98-108
  • DOI: 10.1111/j.1365-2443.2011.01576.x
  • Peer Reviewed
• [Journal Article] Identification of new scavengers for hydroxyl radicals and superoxide dismutase by utilising ultraviolet A photoreaction of 8-methoxysoralen and a variety of mutants of Escherichia coli: implications on certain diseases of DNA repair deficiency. (2012)
  • Author(s): Ahmad SI
  • Journal Title: J Photochem Photobiol B Volume: 116 Pages: 30-36
  • Peer Reviewed
• [Journal Article] The vital role of pol ζ and REV1 in mutagenic, but not correct, DNA synthesis across benzo[α]pyrene-dG and the recruitment of pol ζ by REV1 to a replication-stalled site (2012)
  • Author(s): Hashimoto K
  • Journal Title: J.Biol. Chem. Volume: 287 Pages: 9613-9622
  • DOI: 10.1074/jbc.m111.331728
  • Peer Reviewed
• [Journal Article] The architecture of the CRL4^DDB2/CSA ubiquitin ligase, targeting and release of COP9 signalsome inhibition. (2011)
  • Author(s): Fischer, E.S., et al
  • Journal Title: Cell Volume: 147 Pages: 1024-1039
  • DOI: 10.1016/j.cell.2011.10.035
  • Peer Reviewed
• [Journal Article] NBS1 recruites RAD18 via a RAD6-like domain and regulates Polη- dependent transleion 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 CM, Mori T, Zou L, Komatsu K.
  • Journal Title: Mol. Cell Volume: 43 Pages: 788-797
  • DOI: 10.1016/j.molcel.2011.07.026
  • NAID: 120003338825
  • Peer Reviewed
• [Journal Article] Molecular chaperon Hsp90 regulates REV1- mediated mutagenesis. Mol (2011)
  • Author(s): Pozo FM
  • Journal Title: Cell Biol Volume: 31 Pages: 3396-3409
  • DOI: 10.1128/mcb.05117-11
  • Peer Reviewed
• [Journal Article] Hst3 and Hst4 histone deacetylases regulate replicative lifespan by preventing genome instability in Saccharomyces cerevisiae (2011)
  • Author(s): Hachinohe, M., Hanaoka, F., and Masumoto, H
  • Journal Title: Genes Cells Volume: 16 Pages: 467-477
  • DOI: 10.1111/j.1365-2443.2011.01493.x
  • Peer Reviewed
• [Journal Article] Genetic analysis using a mouse cell cycle mutant identifies magoh as a novel gene involved in Cdk regulation (2011)
  • Author(s): Inaki, M., Kato, D., Utsugi, T., Onoda, F., Hanaoka, F., and Murakami, Y
  • Journal Title: Genes Cells Volume: 16 Pages: 166-178
  • DOI: 10.1111/j.1365-2443.2010.01479.x
  • Peer Reviewed
• [Journal Article] Photosensitized [2 + 2] cycloaddition on N-acetylated cytosine affords stereoselective formation of cyclobutane pyrimidine dimer (2011)
  • Author(s): Yamamoto, J., Nishiguchi, K., Manabe, K., Masutani, C., Hanaoka, F., and Iwai, S
  • Journal Title: Nucleic Acid Res Volume: 39 Pages: 1165-1175
  • DOI: 10.1093/nar/gkq855
  • Peer Reviewed
• [Journal Article] Photosensitized [2+2] cycloaddition of N-acetylated cytosine affords stereoselective formation of cyclobutane pyrimidine dimer (2011)
  • Author(s): Yamamoto, J., et al
  • Journal Title: Nucl.Acids Res. Volume: 39 Pages: 1165-1175
  • Peer Reviewed
• [Journal Article] Simultaneous disruption of two DNA polymerases, Polηand Polζin avian DT40 cells unmasks the role of Polηin celluler 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., and Takeda, S
  • Journal Title: PLoS Genet Volume: 6, pii
  • DOI: 10.1371/journal.pgen.1001151
  • NAID: 120002661519
  • Peer Reviewed
• [Journal Article] Characterization of a Y-family DNA polymeraseηfrom the eukaryotic thermophile Alvinella pompejana (2010)
  • Author(s): Kashiwagi, S., Kuraoka, I., Fujiwara, Y., Hitomi, K., Cheng, Q. J., Shin, D. S., Masutani, C., Tainer, J. A., Hanaoka, F., and Iwai, S
  • Journal Title: J. Nucleic Acids pii Pages: 701472-701472
  • DOI: 10.4061/2010/701472
  • 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 pii Pages: 805698-805698
  • DOI: 10.4061/2010/805698
  • NAID: 120003202349
  • Peer Reviewed
• [Journal Article] Structure and mechanism of human DNA polymeraseη (2010)
  • Author(s): Biertümpfel, C., Zhao, Y., Kondo, Y., Ramon-Maiques, S., Gregory, M., Lee, J. Y., Masutani, C., Lehmann, A. R., Hanaoka, F., and Yang, W
  • Journal Title: Nature Volume: 465 Pages: 1044-1049
  • DOI: 10.1038/nature09196
  • Peer Reviewed
• [Journal Article] Structure and mutagenesis studies of the C-terminal region of licensing factor Cdt1 enable the identification of key residues for binding to replicative helicase Mcm proteins (2010)
  • Author(s): Jee, J.-G., Mizuno, T., Kamada, K., Tochio, T., Chiba, Y., Yanagi, K., Yasuda, G., Hiroaki, H., Hanaoka, F. and Shirakawa, M
  • Journal Title: J. Biol. Chem Volume: vol.285(21) Pages: 15931-15940
  • DOI: 10.1074/jbc.m109.075333
  • Peer Reviewed
• [Journal Article] Characterization of a Y-family DNA polymerase eta from the eukaryotic thermophile Alvinella pompejana (2010)
  • Author(s): Kashiwagi, S., et al
  • Journal Title: J Nucl.Acids Volume: 2010
  • Peer Reviewed
• [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., et al
  • Journal Title: PLoS Genetics Volume: 6
  • Peer Reviewed
• [Presentation] 損傷トレランスと突然変異生成に働く乗り越え複製ポリメラーゼ (2014)
  • Author(s): 花岡文雄
  • Organizer: 日本薬学会第134年会特別講演
  • Place of Presentation: 熊本
  • Invited
• [Presentation] Defective translesion DNA synthesis and links to human disease (2014)
  • Author(s): Hanaoka, F
  • Organizer: International Symposium on Xeroderma Pigmentosum and Related Diseases : Disorders of DNA Damage Response–Bench to Bedside-
  • Place of Presentation: 神戸
  • Invited
• [Presentation] (2014)
  • Author(s): Osakabe, A., Tachiwana, H., Horikoshi, N., Kagawa, W., Yasuda, T., Hanaoka, F., Sugasawa, K., Iwai, S., and Kurumizaka, H
  • Organizer: International Conference, Kyoto, 2014: Replication, Repair and Recombination; coupling mechanisms and chromatin dynamics for genome integrity
  • Place of Presentation: 京都
  • Invited
• [Presentation] All road leads to DNA-lesion bypass : mechanisms of translesion synthesis (2014)
  • Author(s): Yang, W., Lee, Young-Sam, Zhao, Y., Nakamura, T., Biertümpfel, C., Yamagata, Y., Hua, Y., and Hanaoka, F
  • Organizer: International Conference, Kyoto, 2014: Replication, Repair and Recombination; coupling mechanisms and chromatin dynamics for genome integrity
  • Place of Presentation: 京都
  • Invited
• [Presentation] Physiological role of TLS polymerases in mouse skin upon UV irradiation (2014)
  • Author(s): Yokoi, M., Sakurai, Y., Hando, N., Morita, D., and Hanaoka, F
  • Organizer: International Conference, Kyoto, 2014: Replication, Repair and Recombination; coupling mechanisms and chromatin dynamics for genome integrity
  • Place of Presentation: 京都
  • Invited
• [Presentation] Regulation of DNA damage tolerance distinct from Polη-mediated translesion synthesis (2014)
  • Author(s): Kanao, R., Masuda, Y., Hanaoka, F., and Masutani, C
  • Organizer: International Conference, Kyoto, 2014: Replication, Repair and Recombination; coupling mechanisms and chromatin dynamics for genome integrity
  • Place of Presentation: 京都
  • Invited
• [Presentation] Role of DNA polymerase eta in somatic hypermutation (2013)
  • Author(s): Hanaoka, F
  • Organizer: The 3^<rd> Frence-Japanese Cancer Meeting
  • Place of Presentation: Toulouse, France
  • Invited
• [Presentation] Structure and function of mammalian DNA polymerase eta with special reference on translesion synthesis and somatic hypermutation (2013)
  • Author(s): Hanaoka, F
  • Organizer: 京都大学大学院医学研究科・免疫ゲノム医学特別セミナー
  • Place of Presentation: 京都
  • Year and Date: 2013-05-27
  • Invited
• [Presentation] DNAポリメラーゼ・イータ—その構造と多彩な機能— (2013)
  • Author(s): 花岡文雄
  • Organizer: 公開シンポジウム「遺伝情報場:構築を担う分子のダイナミクスと制御」
  • Place of Presentation: 東京
  • Year and Date: 2013-01-11
  • Invited
• [Presentation] Functional roles of mammalian DNA polymerase eta (2012)
  • Author(s): Hanaoka, F
  • Organizer: Second Japanese-French Cancer Workshop
  • Place of Presentation: 鳴門
  • Invited
• [Presentation] Alternative translesion synthesis pathway via interaction between stalled DNA polymeraseηand REV1 (2012)
  • Author(s): Hanaoka, F., Yokoi, M
  • Organizer: 8th 3R Symposium
  • Place of Presentation: 淡路島
  • Invited
• [Presentation] Functional roles of mammalian DNA polymeraseη, the product of the XP-V responsible gene (2012)
  • Author(s): Hanaoka, F., Yokoi, M
  • Organizer: 3rd Erling Seeberg Symposium
  • Place of Presentation: Trondheim and Orland, Norway
  • Invited
• [Presentation] Physiological role of DNA polymeraseη-REV1 interaction in mammalian cells (2012)
  • Author(s): Yokoi, M., Hanaoka, F
  • Organizer: Cantoblanco Workshops : Polymerases Involved in DNA Replication, Repair and Mutagenesis
  • Place of Presentation: Madrid, Spain
  • Invited
• [Presentation] Involvement of DNA polymeraseη-REV1 interaction in a part of error-prone translesion DNA synthesis (2012)
  • Author(s): Hanaoka, F., Yokoi, M
  • Organizer: US-Japan DNA Repair Workshop
  • Place of Presentation: Leesburg, USA
  • Invited
• [Presentation] Involvement of DNA polymerase eta-REV1 interaction in a part of error-prone translesion DNA synthesis. (2012)
  • Author(s): Hanaoka F
  • Organizer: The 4th U.S.-Japan DNA Repair Meeting
  • Place of Presentation: Virginia, USA
  • Invited
• [Presentation] Functional roles of mammalian DNA polymerase eta, the product of the XP-V responsible gene. (2012)
  • Author(s): Hanaoka F
  • Organizer: The 3rd Erling Seeberg Symposium on DNA Repair
  • Place of Presentation: Trondheim and Orland, Norway
  • Invited
• [Presentation] Alternative translesion synthesis pathway via interaction between stalled DNA polymerase eta and REV1 (2012)
  • Author(s): Hanaoka F
  • Organizer: The 8th 3R Symposium together with National Institute of Genetics “2012 International Symposium”: “Molecular Mechanisms and Pathology of the 3R”
  • Place of Presentation: 淡路市、兵庫
  • Invited
• [Presentation] Functional roles of mammalian DNA polymerase eta (2012)
  • Author(s): Hanaoka F
  • Organizer: 2nd Japanese-Frence Cancer Workshop
  • Place of Presentation: 鳴門市、徳島
  • Invited
• [Presentation] 突然変異、損傷乗り越え複製 (2012)
  • Author(s): 花岡文雄.能美先生
  • Organizer: 能美健彦先生ご退官記念祝賀会
  • Place of Presentation: 東京
  • Year and Date: 2012-07-07
  • Invited
• [Presentation] Structure, mechanism and functions of mammalian DNA polymerase eta (2012)
  • Author(s): Hanaoka, F
  • Organizer: Special Seminar at the University of Texas MD Anderson Cancer Center
  • Place of Presentation: Austin, USA
  • Year and Date: 2012-04-10
  • Invited
• [Presentation] Biochemical properties and structure of human DNA polymerase &eta, xeroderma pigmentosum variant- responsible gene product (2011)
  • Author(s): Hanaoka, F., Kondo, Y., Masutani, C., Biertümpfel, C., Zhao, Y., Yang, W
  • Organizer: GDRI France Japan Conference
  • Place of Presentation: Montpellier, France
  • Invited
• [Presentation] DNA修復研究における生化学的アプローチ (2011)
  • Author(s): 花岡文雄
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
  • Invited
• [Presentation] DNA修復と疾患:色素性乾皮症を中心に (2011)
  • Author(s): 花岡文雄
  • Organizer: 第28回日本医学会総会
  • Place of Presentation: 東京
  • Invited
• [Presentation] Biochemical properties and structure of human DNA polymerase η. (2011)
  • Author(s): Hanaoka, F.
  • Organizer: 6th UK-Japan Cell Cycle Workshop
  • Place of Presentation: Lake Windermere, UK
  • Invited
• [Presentation] Biochemical properties and structure of human DNA polymerase &eta, xeroderma pigmentosum variant-responsible gene product (2011)
  • Author(s): Hanaoka, F.
  • Organizer: GDRI France Japan Conference
  • Place of Presentation: Montpellier, France
  • Invited
• [Presentation] Regulation of translesion DNA synthesis with special emphasis on DNA polymerase eta and PCNA ubiquitylation (2010)
  • Author(s): Hanaoka, F., Deguchi, S., Kanao, R., and Masutani, C
  • Organizer: The EMBO Workshop on"Interface between the Ubiquitin Family and the DNA Damage Response"
  • Place of Presentation: Rovinj, Croatia
  • Invited
• [Presentation] Tumorigenesis induced by chronic treatment with UV-B in Pol eta- and Xpc-deficient mice (2010)
  • Author(s): Hanaoka, F
  • Organizer: Gordon Research Conference on Mutagenesis
  • Place of Presentation: Waterville, Maine, USA
  • Invited
• [Presentation] Regulation of translesion DNA synthesis with special emphasis on DNA polymerase eta and PCNA ubiquitylation (2010)
  • Author(s): 花岡文雄
  • Organizer: EMBOワークショップ「ユビキチンファミリーとDNA損傷応答の関わり」
  • Place of Presentation: ロビーニ、クロアチア(招待講演)
  • Year and Date: 2010-09-04
• [Presentation] 遺伝子の傷を治して発がんを抑える (2010)
  • Author(s): 花岡文雄
  • Organizer: 大切な「いのち」を守る科学学際生命科学東京コンソーシアム市民講演会平成21年度文部科学省・戦略的大学連携支援プログラム採択
  • Place of Presentation: 東京
  • Year and Date: 2010-04-10
  • Invited
• [Presentation] DNAポリメラーゼ・イータ ―その構造と多彩な機能―
  • Author(s): 花岡文雄
  • Organizer: 新学術領域研究「遺伝情報場」公開シンポジウム
  • Place of Presentation: 東京
  • Invited
• [Remarks]
  • URL: http://www-cc.gakushuin.ac.jp/~20080213/
• [Remarks]
  • URL: http://www-cc.gakushuin.ac.jp/~20080213/