Analysis of Homologous DNA Recombination using the chicken DT40 B cell line

• Project/Area Number: 17390076
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Kyoto University
• Principal Investigator: TAKEDA Shunichi KYOTO UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, PROFESSOR, 医学研究科, 教授 (60188191)
• Co-Investigator(Kenkyū-buntansha): 竹中 克也 京都大学, 医学研究科, 21世紀COE特任助手 (20378706)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥14,200,000 (Direct Cost: ¥14,200,000); Fiscal Year 2006: ¥11,200,000 (Direct Cost: ¥11,200,000); Fiscal Year 2005: ¥3,000,000 (Direct Cost: ¥3,000,000)
• Keywords: Homologous Recombination / Targeted integration / DT40 cell / Gene targeting / Rad18 / Ubc13 / Fbh1 / DNA polymerase / 遺伝学 / 癌 / 細胞・組織 / 放射線 / 老化

Research Abstract

In higher eukaryotic cells, transfected DNA is occasionally integrated into the genome predominantly through random integration. A novel exception of this rule is chicken B lymphocyte lines, including DT40 cells, in which random integration and targeted one occur with similar frequencies. The goal of this project is to identify the molecular mechanisms underlying the efficient gene targeting in DT40 cells. While the mechanisms have not yet clarified, we have revealed a role of some factors in Homologous DNA Recombination (HR), as delineated below.
(1) Yeast Rad18, E3 ubiquitin ligase is essential for postreplicational repair (PRR), but is not involved in HR. We found that Rad18 suppresses the toxic effect of nonhomolgous end-joining, and thereby facilitates HR (Saberi et al., 2007).
(2) Ubc13, E3 ubiquitin ligase is involved in a type of PRR, but not in HR in lower eukaryotes. We found that Ubc13 plays a critical role in an initial step of HR dependent DSB repair (Zhao et al., 2007).
(3) FBH1 seems to prevent aberrant HR by suppressing the assembly of Rad51 at DNA damage in the fission yeast. In DT40 cells, we found that FBH1 does not affect Rad51 accumulation, but acts in parallel with Bloom helicase to suppress HR at a late step (Kohzaki et al., 2007).

Research Products

• [Journal Article] A critical role for the ubiquitin-conjugating enzyme Ubc13 in initiating homologous recombination. (2007)
  • Author(s): Zhao GY, Sonoda E, Barber LJ, Oka H, Murakawa Y, Yamada K, Ikura T, Wang X, Kobayashi M, Yamamoto K, Boulton SJ, Takeda S
  • 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] 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 NV, Szuts D, Sale JE, 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] RAD18 and poly[ADP ribose]polymerase independently suppress the access of nonhomologous end joining to double strand breaks and facilitate homologous recombination mediated repair. (2007)
  • Author(s): Saberi A, Hochegger H, Szuts D, Lan L, Yasui A, Sale JE, Taniguchi Y, Murakawa Y, Hua W, Yokomori K, Helleday T, Teraoka H, Arakawa H, Buerstedde JM, Takeda S
  • Journal Title: Mol Cell Biol 27 Pages: 2562-2571
  • 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 GY, Sonoda E, Barber LJ, Oka H, Murakawa Y, Yamada K, Ikura T, Wang X, Kobayashi M, Yamamoto K, Boulton SJ, Takeda S.
  • 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] 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 NV, Szuts D, Sale JE, 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] RAD18 and poly[ADP ribose]polymerase independently suppress the access of nonhomologous end joining to double strand breaks and facilitate homologous recombination mediated repair. (2007)
  • Author(s): Saberi A, Hochegger H, Szuts D, Lan L, Yasui A, Sale JE, Taniguchi Y, Murakawa Y, Hua W, Yokomori K, Helleday T, Teraoka H, Arakawa H, Buerstedde JM, Takeda S.
  • Journal Title: Mol Cell Biol. 27 Pages: 2562-2571
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A critical role for the ubiquitin-conjugating enzyme Ubc13 in initiating homologuous recombination. (2007)
  • Author(s): Zhao GY, Sonoda E, Barber LJ, Oka H, Murakawa Y, Yamada K, Ikura T, Wang X, Kobayashi M, Yamamoto K, Boulton SJ, Takeda S
  • Journal Title: Mol Cell 25 Pages: 663-675
• [Journal Article] RAD18 and poly[ADP ribose]polymerase independently suppress the access of nonhomologous end joining to double strand breaks and facilitate homologous recombination mediated repair. (2007)
  • Author(s): Saberl A, Hochegger H, Szuts D, Lan L, Yasul A, Sale JE, Taniguchi Y, Murakawa Y, Hua W, Yokomori K, Helleday T, Teraoka H, Arakawa H, Buerstedde JM, Takeda S
  • Journal Title: Mol Cell Biol 27 Pages: 2562-2571
• [Journal Article] Involvement of vertebrate Polkappa in translesion DNA synthesis across DNA monoalkylation damage. (2006)
  • Author(s): Takenaka T, Ogi T, Okada T, Sonoda E, Guo C, Friedberg EC, 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] Genetic dissection of vertebrate 53BP1 : a major role in non-homologous end joining of DNA double strand breaks. (2006)
  • Author(s): Nakamura K, Sakai W, Kawamoto T, Bree RT, Lowndes NF, Takeda S, Taniguchi Y
  • Journal Title: DNA Repair (Amst) 5 Pages: 741-749
  • Description: 「研究成果報告書概要(和文)」より
• [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 TS, Parker JL, Bielen AB, Takeda S, Ulrich HD, Friedberg EC
  • Journal Title: Mol Cell 23 Pages: 265-271
  • 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] Ubiquitin-Binding Motifs in REV1 Protein Are Required For Its Role in the Tolerance of DNA Damage. (2006)
  • Author(s): Guo C, Tang TS, Bienko M, Parker JL, Bielen AB, Sonoda E, Takeda S, Ulrich HD, Dikic I, Friedberg EC
  • 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] Vertebrate POLQ and POL β cooperate in Base Excision Repair of Oxidative DNA Damage. (2006)
  • Author(s): Yoshimura A, Kohzaki M, Nakamura J, Asagoshi K, Sonoda E, Hou E, Prasad R, Wilson SH, Tano K, Yasui A, Lan L, Seki M, Wood RD, Arakawa H, Buerstedde JM, 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(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 EC, Takeda S.
  • Journal Title: J Biol Chem. 281 Pages: 2000-2004
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genetic dissection of vertebrate 53BP1 : a major role in non-homologous end joining of DNA double strand breaks. (2006)
  • Author(s): Nakamura K, Sakai W, Kawamoto T, Bree RT, Lowndes NF, Takeda S, Taniguchi Y.
  • Journal Title: DNA Repair (Amst) 5 Pages: 741-749
  • Description: 「研究成果報告書概要(欧文)」より
• [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 TS, Parker JL, Bielen AB, Takeda S, Ulrich HD, Friedberg EC.
  • Journal Title: Mol Cell. 23 Pages: 265-271
  • Description: 「研究成果報告書概要(欧文)」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(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 TS, Bienko M, Parker JL, Bielen AB, Sonoda E, Takeda S, Ulrich HD, Dikic I, Friedberg EC.
  • 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 POLβ cooperate in Base Excision Repair of Oxidative DNA Damage. (2006)
  • Author(s): Yoshimura A, Kohzaki M, Nakamura J, Asagoshi K, Sonoda E, Hou E, Prasad R, Wilson SH, Tano K, Yasui A, Lan L, Seki M, Wood RD, Arakawa H, Buerstedde JM, 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] Ubiquitin-Binding Motifs in REV1 Protein Are Required For Its Role in the Tolerance of DNA Damage. (2006)
  • Author(s): Guo C, Tang TS, Blenko M, Parker JL, Bielen AB, Sonoda E, Takeda S, Ulrich HD, Dikic I, Friedberg EC
  • Journal Title: Mol Cell Biol 26 Pages: 8892-8900
• [Journal Article] Vertebrate POLQ and POL β cooperate in Base Excision Repair of Oxidative DNA Damage. Mol Cell. 24 : 115-25 (2006)
  • Author(s): Yoshimura A, Kohzakl M, Nakamura J, Asagoshi K, Sonoda E, Hou E, Prasad R, Wilson SH, Tano K, Yasul A, Lan L, Sekl M, Wood RD, Arakawa H, Buerstedde JM, Hochegger H, Okada T, Hiraoka M, Takeda S
  • Journal Title: Mol Cell 24 Pages: 115-125
• [Journal Article] Involvement of vertebrate Polkappa in translesion DNA synthesis across DNA monoalkylation damage. (2006)
  • Author(s): Takenaka T, Ogi T, Okada T, Sonoda E, Guo C, Friedberg EC, Takeda S
  • Journal Title: Journal of Biological Chemistry 281 Pages: 2000-2004
• [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, Kozaki M, Takeda S
  • Journal Title: Mol Cell Biol 25 Pages: 6103-6111
  • Description: 「研究成果報告書概要(和文)」より
• [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, Kozaki M, Takeda S.
  • Journal Title: Mol Cell Biol. 25 Pages: 6103-6111
  • Description: 「研究成果報告書概要(欧文)」より
• [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, Kozaki M, Takeda S
  • Journal Title: Molecular Cell Biology 25 Pages: 6103-6111