Bio-imaging of DNA repair machinery

• Project/Area Number: 17570160
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Cell biology
• Research Institution: Kyoto University
• Principal Investigator: SONODA Eiichiro KYOTO UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, ASSOCIATE PROFESSOR, 医学研究科, 助教授 (50281093)
• Co-Investigator(Kenkyū-buntansha): TANO Keizo KYOTO UNIVERSITY, RESEARCH REACTOR INSTITUTE, ASSOCIATE PROFESSOR, 原子炉実験所, 助教授 (00183468)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2006: ¥2,400,000 (Direct Cost: ¥2,400,000); Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
• Keywords: DT40 / DSB repair / UBC13 / Polymerase Q / Ubiqutine / Laser induced damage / homologous recombination / translesion synthesis

Research Abstract

To elucidate the dynamic process of DNA repair, we employed GFP-fusion technique and immmuno-cytochemistry combined with confocal laser microscope. We generated POLQ-GFP and expressed in human HeLa cells and found that PolQ accumulate at the oxidative DNA damage site induced by Laser. Further biochemical analysis revealed that PolQ bypass abasic sites in vitro. Taken together, we conclude that PolQ is a translesion polymerase involved in base excision repair. UBC13 is a ubiqutin conjugating enzyme whose function is unknown in vivo. We found that UBC13 is critical for DNA double strand break repair by facilitating the formation of DNA damage repair foci containing various repair proteins. In UBC13 deficient cells, accumulation of Rad51, RPA, Brcal to the damage sites are severely impaired, probably due to the insufficient processing of double strand break ends. We conclude that UBC13 facilitates double strand break repair mediated by homologous recombination. Ubiquitination of unknown substrate including histone proteins appear to play a critical role for generating the repair substrates for homologous recombination.

Research Products

• [Journal Article] Interplay between DNA polymerases beta and lambda in repair of oxidation DNA damage in chicken DT40 cells (2007)
  • Author(s): Tano, K., Nakamura, J., Asagoshi, K., Arakawa, H., Sonoda, E., Braithwaite, E.K., Prasad, R., Buerstedde, J.M., Takeda, S., Watanabe, M., Wilson, S.H.
  • Journal Title: DNA Repair (Amst) (In press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Collaborative roles of gammaH2AX and the Rad51 paralog Xrcc3 in homologous recombinational repair. (2007)
  • Author(s): Sonoda, E., Zhao, G.Y., Kohzaki, M., Dhar, P.K., Kikuchi, K., Redon, C., Pilch, D.R., Bonner, W.M., Nakano, A., Watanabe, M., Nakayama, T., et al.
  • Journal Title: DNA Repair (Amst) 6 Pages: 280-292
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cooperative Roles of Vertebrate Fbh1 and BIm DNA Helicases in Avoidance of Crossovers during Recombination Initiated by Replication Fork Collapse. (2007)
  • Author(s): Kohzaki, M., Hatanaka, A., Sonoda, E., Yamazoe, M., Kikuchi, K., Trung, N.V., Szuts, D., Sale, J.E., Shinagawa, H., Watanabe, M., Takeda, S.
  • Journal Title: Mol Cell Biol 27 Pages: 2812-2820
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A Critical Role for the Ubiquitin-Conjugating Enzyme Ubc13 in Initiating Homologous Recombination. (2007)
  • Author(s): Zhao, G.Y., Sonoda, E., Barber, L.J., Oka, H., Murakawa, Y., Yamada, K., Ikura, T., Wang, X., Kobayashi, M., Yamamoto et al.
  • Journal Title: Mol Cell 25 Pages: 663-675
  • NAID: 130007000910
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Critical Role for the Ubiquitin-Conjugating Enzyme Ubc13 in Initiating Homologous Recombination. (2007)
  • Author(s): Zhao, G.Y., Sonoda, E., Barber, L J., Oka, H., Murakawa, Y., Yamada, K., Ikura, T., Wang, X., Kobayashi, M., Yamamoto, K., Boulton, S.J., Takeda, S.A
  • Journal Title: Mol Cell 25 Pages: 663-675
  • NAID: 130007000910
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Interplay between DNA polymerases beta and lambda in repair of oxidation DNA damage in chicken DT40 cells. (2007)
  • Author(s): Tano, K., Nakamura, J., Asagoshi, K., Arakawa, H., Sonoda, E., Braithwaite, E.K., Prasad, R., Buerstedde, J.M., Takeda, S., Watanabe, M., Wilson, S.H.
  • Journal Title: DNA Repair (Amst)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cooperative Roles of Vertebrate Fbh1 and Blm DNA Helicases in Avoidance of Crossovers during Recombination Initiated by Replication Fork Collapse (2007)
  • Author(s): Kohzaki, M., Hatanaka, A., Sonoda, E., Yamazoe, M., Kikuchi, K., Vu Trung, N., Szuts, D., Sale, J.E., Shinagawa, H., Watanabe, M., Takeda, S.
  • Journal Title: Mol Cell Biol 27 Pages: 2812-2820
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] naturally occurring genetic variant of human XRCC2 (R188H) confers increased resistance to cisplatin-induced DNA damage (2007)
  • Author(s): Danoy, P., Sonoda, E., Lathrop, M., Takeda, S., Matsuda, F.A
  • Journal Title: Biochem Biophys Res Commun 352 Pages: 763-768
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cooperative Roles of Vertebrate Fbh1 and Blm DNA Helicases in Avoidance of Crossovers during Recombination Initiated by Replication Fork Collapse. (2007)
  • Author(s): Kohzaki, M., Hatanaka, A., Sonoda, E., Yamazoe, M., Kikuchi, K., Trung, N.V., Szuts, D., Sale, J.E., Shinagawa, H., Watanabe, M., Takeda, S.
  • Journal Title: Mol Cell Biol 27 Pages: 2812-2820
• [Journal Article] REV1 Protein Interacts with PCNA : Significance of the REV1 BRCT Domain In Vitro and In Vivo. (2006)
  • Author(s): Guo, C., Sonoda, E., Tang, T.S., Parker, J.L., Bielen, A.B., Takeda, S., Ulrich, H.D., Friedberg, E.C.
  • Journal Title: Mol Cell 23 Pages: 265-271
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Ubiquitin-Binding Motifs in REV1 Protein Are Required For Its Role in the Tolerance of DNA Damage. (2006)
  • Author(s): Guo, C., Tang, T.S., Bienko, M., Parker, J.L., Bielen, A.B., Sonoda, E., Takeda, S., Ulrich, H.D., Dikic, I., Friedberg, E.C.
  • Journal Title: Mol Cell Biol 26 Pages: 8892-8900
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Parp-1 protects homologous recombination from interference by Ku and Ligase IV in vertebrate cells. (2006)
  • Author(s): Hochegger, H., Dejsuphong, D., Fukushima, T., Morrison, C., Sonoda, E., Schreiber, V., Zhao, G.Y., Saberi, A., Masutani, M., Adachi, N., Koyama, H., de Murcia, G., Takeda, S.
  • Journal Title: Embo J 25 Pages: 1305-1314
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Differential usage of non-homologous end-joining and homologous recombination in double strand break repair. (2006)
  • Author(s): Sonoda, E., Hochegger, H., Saberi, A., Taniguchi, Y., Takeda, S.
  • Journal Title: DNA Repair (Amst) 5 Pages: 1021-1029
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] REV3 and REV1 play major roles in recombination-independent repair of DNA interstrand cross-links mediated by monoubiquitinated proliferating cell nuclear antigen (PCNA). (2006)
  • Author(s): Shen, X., Jun, S., O'Neal, L.E., Sonoda, E., Bemark, M., Sale, J.E., Li, L.
  • Journal Title: J Biol Chem 281 Pages: 13869-13872
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Involvement of vertebrate Polkappa in translesion DNA synthesis across DNA monoalkylation damage. (2006)
  • Author(s): Takenaka, K., Ogi, T., Okada, T., Sonoda, E., Guo, C., Friedberg, E.C., Takeda, S.
  • Journal Title: J Biol Chem 281 Pages: 2000-2004
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Critical roles for polymerase zeta in cellular tolerance to nitric oxide-induced DNA damage. (2006)
  • Author(s): Wu, X., Takenaka, K., Sonoda, E., Hochegger, H., Kawanishi, S., Kawamoto, T., Takeda, S., Yamazoe, M.
  • Journal Title: Cancer Res 66 Pages: 748-754
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Vertebrate POLQ and POLbeta Cooperate in Base Excision Repair of Oxidative DNA Damage. (2006)
  • Author(s): Yoshimura, M., Kohzaki, M., Nakamura, J., Asagoshi, K., Sonoda, E., Hou, E., Prasad, R., Wilson, S.H., Tano, K., Yasui, A., Lan, L., Seki, M., Wood, R.D., Arakawa, H., Buerstedde, J.M., Hochegger, et al.
  • Journal Title: Mol Cell 24 Pages: 115-125
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A novel Rad18 function involved in protection of the vertebrate genome after exposure to camptothecin. (2006)
  • Author(s): Yoshimura, A., Nishino, K., Takezawa, J., Tada, S., Kobayashi, T., Sonoda, E., Kawamoto, T., Takeda, S., Ishii, Y., Yamada, K., Enomoto, T., Seki, M.
  • Journal Title: DNA Repair (Amst) 5 Pages: 1307-1316
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A naturally occurring genetic variant of human XRCC2 (R188H) confers increased resistance to cisplatin-induced DNA damage. (2006)
  • Author(s): Danoy, P., Sonoda, E., Lathrop, M., Takeda, S., Matsuda, F.
  • Journal Title: Biochem Biophys Res Commun 352 Pages: 763-768
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] novel Rad18 function involved in protection of the vertebrate genome after exposure to camptothecin. (2006)
  • Author(s): Yoshimura, A., Nishino, K., Takezawa, J., Tada, S., Kobayashi, T., Sonoda, E., Kawamoto, T., Takeda, S., Ishii, Y., Yamada, K., Enomoto, T., Seki, M.A
  • Journal Title: DNA Repair (Amst) 5 Pages: 1307-1316
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Vertebrate POLQ and POLbeta Cooperate in Base Excision Repair of Oxidative DNA Damage (2006)
  • Author(s): Yoshimura, M., Kohzaki, M., Nakamura, J., Asagoshi, K., Sonoda, E., Hou, E., Prasad, R., Wilson, S.H., Tano, K., Yasui, A., Lan, L, Seki, M., Wood, R.D., Arakawa, H., Buerstedde, J.M., Hochegger, H., Okada, T., Hiraoka, M., Takeda, S.
  • Journal Title: Mol Cell 24 Pages: 115-125
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] L REV3 and REV1 play major roles in recombination-independent repair of DNA interstrand cross-links mediated by monoubiquitinated proliferating cell nuclear antigen (PCNA). (2006)
  • Author(s): Shen, X., Jun, S., O'Neal, L.E., Sonoda, E., Bemark, M., Sale, J.E., Li
  • Journal Title: J Biol Chem 281 Pages: 13869-13872
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Ubiquitin-binding motifs in REV1 protein are required for its role in the tolerance of DNA damage. (2006)
  • Author(s): Guo, C., Tang, T.S., Bienko, M., Parker, J.L, Bielen, A.B., Sonoda, E., Takeda, S., Ulrich, H.D., Dikic, I., Friedberg, E.C.
  • Journal Title: Mol Cell Biol 26 Pages: 8892-8900
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Vertebrate POLQ and POLbeta Cooperate In Base Excision Repair of Oxidative DNA Damage. (2006)
  • Author(s): Yoshimura, M., Kohzaki, M., Nakamura, J., Asagoshi, K., Sonoda, E., Hou, E., Prasad, R., Wilson, S.H., Tano, K., Yasui, A., Lan, L., Seki, M., Wood, R.D., Arakawa, H., Buerstedde, J.M., Hochegger, et al.
  • Journal Title: Mol Cell 24 Pages: 115-125
• [Journal Article] A novel Rad18 function involved in protection of the vertebrate genome after exposure to camptothecin. (2006)
  • Author(s): Yoshimura, A., Nishino, K., Takezawa, J., Tada, S., Kobayashi, T., Sonoda, E., Kawamoto, T., Takeda, S., Ishii, Y., Yamada, K., Enomoto, T., Seki.M.
  • Journal Title: DNA Repair (Amst) 5 Pages: 1307-1316
• [Journal Article] Critical roles for polymerase zeta in cellular tolerance to nitric oxide-induced DNA damage (2006)
  • Author(s): Wu, X., Takenaka, K., Sonoda, E., Hochegger, H., Kawanishi, S., Kawamoto, T., Takeda, S., Yamazoe, M.
  • Journal Title: Cancer Res 66 Pages: 748-754
• [Journal Article] Involvement of Vertebrate Pol{kappa} in Translesion DNA Synthesis across DNA Monoalkylation Damage (2006)
  • Author(s): Takenaka, K., Ogi, T., Okada, T., Sonoda, E., Guo, C., Friedberg, E.C., Takeda, S.
  • Journal Title: J Biol Chem 281 Pages: 2000-2004
• [Journal Article] Parp-1 protects homologous recombination from interference by Ku and Ligase IV in vertebrate cells (2006)
  • Author(s): Hochegger, H., Dejsuphong, D., Fukushima, T., Morrison, C., Sonoda, E., Schreiber, V., Zhao, G.Y., Saberi, A., Masutani, M., Adachi, N.
  • Journal Title: EMBO J (印刷中)
• [Journal Article] Differential and collaborative actions of Rad51 paralog proteins in cellular response to DNA damage (2005)
  • Author(s): Yonetani, Y., Hochegger, H., Sonoda, E., Shinya, S., Yoshikawa, H., Takeda, S., Yamazoe, M.
  • Journal Title: Nucleic Acids Res 25 Pages: 6103-6111
• [Journal Article] Multiple roles of vertebrate REV genes in DNA repair and recombination (2005)
  • Author(s): Okada, T., Sonoda, E., Yoshimura, M., Kawano, Y., Saya, H., Kohzaki, M., Takeda, S.
  • Journal Title: Mol Cell Biol 25 Pages: 6103-6111
• [Journal Article] Multiple repair pathways mediate tolerance to chemotherapeutic cross-linking agents in vertebrate cells (2005)
  • Author(s): Nojima, K., Hochegger, et al., Sonoda, E.
  • Journal Title: Cancer Res 65 Pages: 11704-11711
• [Journal Article] Fen-1 facilitates homologous recombination by removing divergent sequences at DNA break ends (2005)
  • Author(s): Kikuchi, K., Taniguchi, Y., Hatanaka, A., Sonoda, E., Hochegger, H., Adachi, N., Matsuzaki, Y., Koyama, H., van Gent, D.C.
  • Journal Title: Mol Cell Biol 25 Pages: 6948-6955
• [Journal Article] Dual roles for DNA polymerase eta in homologous DNA recombination and translesion DNA synthesis (2005)
  • Author(s): Kawamoto, T., Araki, K., Sonoda, E., Yamashita, Y.M., Harada, K., Kikuchi, K., Masutani, C., Hanaoka, F., Nozaki, K.
  • Journal Title: Mol Cell 20 Pages: 793-799
• [Journal Article] Similar effects of Brca2 truncation and Rad51 paralog deficiency on immunoglobulin V gene diversification in DT40 cells support an early role for Rad51 paralogs in homologous recombination (2005)
  • Author(s): Hatanaka, A., Yamazoe, M., Sale, J.E., Takata, M., Yamamoto, K., Kitao, H., Sonoda, E., Kikuchi, K., Yonetani, Y., Takeda, S.
  • Journal Title: Mol Cell Biol 25 Pages: 1124-1134
• [Book] DT40 handbook (2006)
  • Author(s): Eiichiro Sonoda
  • Total Pages: 477
  • Publisher: Springer
  • Description: 「研究成果報告書概要(和文)」より
• [Book] DT40 handbook (2006)
  • Author(s): Sonoda, E.
  • Publisher: Springer(in press)