Regulation of protein expression using a cell cyclce-specific degron and its application to functional analysis

• Project/Area Number: 23651046
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Risk sciences of radiation/Chemicals
• Research Institution: Kyoto University
• Principal Investigator: TAKATA Minoru 京都大学, 放射線生物研究センター, 教授 (30281728)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2011: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: DNA修復 / DNA損傷チェックポイント / Treslin / Geminin / Cdt1 / 細胞周期特異的デグロン

Research Abstract

FUCCIis a recently developed cell cycle specific fluorescent maker. This is based on the cell cycle dependent degradation of Cdt1 or Geminin proteins induced by polyubiquitination. We reasoned this can be exploited if we fuse a part of Cdt1 or Geminin used in FUCCI with a protein forcontrolingexpression in a cell cycle specific manner. This would be most useful when used in combination with powerful genetics in chicken DT40 cell system. Thus we cloned chicken Cdt1 or Geminin thatcorresponds to part of human Cdt1 or Geminin used for FUCCI. However, we could not achieve high expression levels in wild type DT40 cells. We havecreated a cell line rendered conditionally deficient in the Treslin gene, which is a yeast Sld3 homologessential for intiation of DNA replication.This cell line would be useful to study Treslin function.

Research Products

• [Journal Article] NBS1 directly activates ATR independently of MRE11 and TOPBP1. (2013)
  • Author(s): Kobayashi M, Hayashi N, Takata M, Yamamoto K.
  • Journal Title: Genes Cells. Volume: 18 Issue: 3 Pages: 238-46
  • DOI: 10.1111/gtc.12031
  • Peer Reviewed
• [Journal Article] Vandetanib is effective in EGFR-mutant lung cancer cells with PTEN deficiency. (2013)
  • Author(s): Takeda H, Takigawa N, Ohashi K, Minami D, Kataoka I, Ichihara E, Ochi N, Tanimoto M, Kiura K.
  • Journal Title: Mol Cancer Res Volume: 319 Issue: 2 Pages: 417-423
  • DOI: 10.1158/1541-7786.mcr-12-0401
  • NAID: 120005359097
  • Peer Reviewed
• [Journal Article] DNA robustly stimulates FANCD2 monoubiquitylation in the complex with FANCI (2012)
  • Author(s): Koichi Sato. Kazue Toda, Masamichi Ishiai, Minora Takata, Hitoshi Kurumizaka
  • Journal Title: Nucleic Acids Research Volume: 40 Issue: 10 Pages: 4553-4561
  • DOI: 10.1093/nar/gks053
  • Peer Reviewed
• [Journal Article] Histone chaperone activity of Fanconi anemia proteins, FANCD2 and FANCI, is required for DNA crosslink repair (2012)
  • Author(s): Koichi Sato. Masamichi Ishiai, Kazue Toda, Satoshi Furukoshi, Akihisa Osakabe, Hiroaki Tachiwana, Yoshimasa Takizawa, Wataru Kagawa, Hiroyuki Kitao, Naoshi Dohmae, Chikashi Obuse, Hiroshi Kimura, Minoru Takata, Hitoshi Kurumizaka
  • Journal Title: The EMBO Journal Volume: 31 Issue: 17 Pages: 3524-3536
  • DOI: 10.1038/emboj.2012.197
  • Peer Reviewed
• [Journal Article] Mcm8 and Mcm9 form a complex that functions in homologous recombination induced by interstrand crosslinks (2012)
  • Author(s): K. Nishimura
  • Journal Title: Molecular Cell Volume: 47 Issue: 4 Pages: 511-522
  • DOI: 10.1016/j.molcel.2012.05.047
  • Peer Reviewed
• [Journal Article] A ubiquitin-binding protein, FAAP20, links RNF8-mediated ubiquitination to the Fanconi anemia DNA repair network. (2012)
  • Author(s): Yan Z, Guo R, Paramasivam M, Shen W, Ling C, Fox D 3rd, Wang Y, Oostra AB, Kuehl J, Lee DY, Takata M, Hoatlin ME, Schindler D, Joenje H, de Winter JP, Li L, Seidman MM, Wang W
  • Journal Title: Mol Cell. Volume: 47 Issue: 1 Pages: 61-75
  • DOI: 10.1016/j.molcel.2012.05.026
  • Peer Reviewed
• [Journal Article] Establishment of the DNA repair-defective mutants in DT40 cells. (2012)
  • Author(s): Ishiai M, Uchida E, Takata M.
  • Journal Title: Methods Mol Biol. Volume: 920 Pages: 39-49
  • DOI: 10.1007/978-1-61779-998-3_4
  • NAID: 120004996387
  • ISBN: 9781617799976, 9781617799983
  • Peer Reviewed
• [Journal Article] ATR-Chkl signaling pathway and homologous recombinational repair protect cells from 5-fluorouracil cytotoxicity (2012)
  • Author(s): Fujinaka Y, Matsuoka K, Iimori M, Tuul M, Sakasai R, Yoshinaga K, Saeki H, Morita M, Kakeji Y, Gillespie DA, Yamamoto KI, Takata M, Kitao H, Maehara Y
  • Journal Title: DNA Repair Volume: 1(3) Issue: 3 Pages: 247-58
  • DOI: 10.1016/j.dnarep.2011.11.005
  • Peer Reviewed
• [Journal Article] ATR-ATRIP Kinase Complex Triggers Activation of the Fanconi Anemia DNA Repair Pathway (2012)
  • Author(s): Tomoko Shigechi, Junya Tomida, Koichi Sato, Masahiko Kobayashi, John K.Eykelenboom, Fabio Pessina, Yanbin Zhang, Emi Uchida, Masamichi Ishiai, Noel F.Lowndes, Kenichi Yamamoto, Hitoshi Kurumizaka, Yoshihiko Maehara, Minoru Takata
  • Journal Title: Cancer Research Volume: 72 Issue: 5 Pages: 1149-1156
  • DOI: 10.1158/0008-5472.can-11-2904
  • Peer Reviewed
• [Journal Article] Matched sibling donor stem cell transplantation for Fanconi anemia patients with T-cell somatic mosaicism (2012)
  • Author(s): Yabe M, Shimizu T, Morimoto T, Koike T, Takakura H, Tsukamoto H, Muroi K, Oshima K, Asami K, Takata M, Yamashita T, Kato S, Yabe H
  • Journal Title: Pediatric Transplant Volume: (印刷中) Issue: 4 Pages: 340-345
  • DOI: 10.1111/j.1399-3046.2012.01669.x
  • Peer Reviewed
• [Journal Article] FancJ/Brip1 helicase protects against genomic losses and gains invertebrate cells (2011)
  • Author(s): Kitao, et al
  • Journal Title: Genes to Cells Volume: 16 Issue: 6 Pages: 717-724
  • DOI: 10.1111/j.1365-2443.2011.01523.x
  • Peer Reviewed
• [Journal Article] The involvement of SLX4 in interstrand cross-link repair is regulated by the Fanconi anemia pathway (2011)
  • Author(s): Yamamoto KN, Kobayashi S, Tsuda M, Kurumizaka H, Takata M, Kono K, Jiricny J, Takeda S, Hirota K
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 108 Issue: 16 Pages: 6492-6496
  • DOI: 10.1073/pnas.1018487108
  • NAID: 120003072120
  • Peer Reviewed
• [Journal Article] USP1 deubiquitinase maintains phosphorylated CHK1 by limiting its DDB1-dependent degradation (2011)
  • Author(s): Guervilly JH, Renaud E, Takata M, Rosselli F
  • Journal Title: Hum Mol Genet Volume: 20(11) Issue: 11 Pages: 2171-81
  • DOI: 10.1093/hmg/ddr103
  • Peer Reviewed
• [Journal Article] Predisposition to cancer caused by genetic and functional defects of Mammalian atad5 (2011)
  • Author(s): Bell DW, Sikdar N, Lee KY, Price J C, Chatterjee R, Park HD, Fox J, Ishiai M, Rudd ML, Pollock LM, Fogoros SK, Mohamed H, Hanigan CL ; NISC Comparative Sequencing Program, Zhang S, Cruz P, Renaud G, Hansen NF, Cherukuri PF, Borate B, McManus KJ, Stoepel J
  • Journal Title: PLoS Genet Volume: 7(8) Issue: 8 Pages: e1002245-e1002245
  • DOI: 10.1371/journal.pgen.1002245
  • Peer Reviewed
• [Journal Article] Direct Inhibition of TNF-α Promoter Activity by Fanconi Anemia Prote in FANCD2 (2011)
  • Author(s): Nobuko Matsushita, Yujiro Endo, Koichi Sato, Hitoshi Kurumizaka, Takayuki Yamashita, Minoru Takata, Shigeru Yanagi
  • Journal Title: PLoS One Volume: 6(電子媒体による発表) Issue: 8 Pages: e23324-e23324
  • DOI: 10.1371/journal.pone.0023324
  • Peer Reviewed
• [Journal Article] Formaldehyde catabolism is essential in cells deficient for the Fanconi anemia DNA-repair pathway (2011)
  • Author(s): Rosado IV, Langevin F, Crossan GP, Takata M, Patel KJ
  • Journal Title: Nat Struct Mol Biol Volume: 18(12) Issue: 12 Pages: 1432-4
  • DOI: 10.1038/nsmb.2173
  • Peer Reviewed
• [Journal Article] Fanconi anemia : a disorder defective in the DNA damage response (2011)
  • Author(s): Kitao, Takata
  • Journal Title: International Journal of Hematology Volume: 93 Issue: 4 Pages: 417-424
  • DOI: 10.1007/s12185-011-0777-z
  • Peer Reviewed
• [Journal Article] Fanconi anemia : a disorder defective in the DNA damage response (2011)
  • Author(s): Kitao H, Takata M
  • Journal Title: Int J Hematol Volume: 93(4) Issue: 4 Pages: 417-24
  • DOI: 10.1007/s12185-011-0832-9
  • NAID: 10029534160
  • Peer Reviewed
• [Journal Article] The role of SNM1 family nucleases in etoposide-induced apoptosis (2011)
  • Author(s): Hosono, Y., Abe, T., Ishiai, M., Takata, M., Enomoto, T., Seki, M.
  • Journal Title: Biochem.Biophys.Res.Commun. Volume: 410 Issue: 3 Pages: 568-573
  • DOI: 10.1016/j.bbrc.2011.06.027
  • Peer Reviewed
• [Presentation] DNA損傷シグナルとファンコニ貧血経路 (2012)
  • Author(s): 高田 穣
  • Organizer: 生体調節研究所セミナー
  • Year and Date: 2012-11-15
• [Presentation] Genetic interplay betweenthe Fanconi anemia pathway and aldehyde metabolism in humans (2012)
  • Author(s): Minoru Takata, Asuka Hira, Naoya Suzuki, Akira Niwa, Tatsutoshi Nakahata, Hiromasa Yabe, Megumu K. Saito, Keitaro Matsuo, Miharu Yabe
  • Organizer: The 8th3R Symposium
  • Place of Presentation: 淡路市
• [Presentation] Genetic interplay betweenthe Fanconi anemia pathway and aldehyde metabolism in humans (2012)
  • Author(s): Minoru Takata, Asuka Hira, Naoya Suzuki, Akira Niwa, Tatsutoshi Nakahata, Hiromasa Yabe, Megumu K. Saito, Keitaro Matsuo, Miharu Yabe
  • Organizer: 28th RBC-NIRS INTERNATIONAL SYMPOSIUM
  • Place of Presentation: 京都市
• [Presentation] Molecular pathogenesis of Fanconi anemia (2012)
  • Author(s): Minoru Takata
  • Organizer: 第54回日本小児血液・がん学会学術集会
  • Place of Presentation: 横浜
• [Presentation] Crosstalk signaling between checkpoint and the Fanconi anemia DNA repair pathway (2012)
  • Author(s): Junya Unno, Junya Tomida, Tomoko Shigechi, Akiko Itaya, and Minoru Takata
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌・ホテルさっぽろ芸文館・札幌市教育文化会館（北海道）
  • Invited
• [Presentation] Genetic interplay between the Fanconi anemia pathway and aldehyde metabolism in humans” (2012)
  • Author(s): Minoru Takata, Asuka Hira, Naoya Suzuki, Akira Niwa, Tatsutoshi Nakahata, Hiromasa Yabe, Megumu K. Saito, Keitaro Matsuo, Miharu Yabe
  • Organizer: The 8th 3R Symposium
  • Place of Presentation: 淡路夢舞台国際会議場（兵庫県）
  • Invited
• [Presentation] Genetic interplay between the Fanconi anemia pathway and aldehyde metabolism in humans” (2012)
  • Author(s): Minoru Takata, Asuka Hira1, Naoya Suzuki, Akira Niwa, Tatsutoshi Nakahata, Hiromasa Yabe, Megumu K. Saito, Keitaro Matsuo, Miharu Yabe
  • Organizer: 28th RBC-NIRS INTERNATIONAL SYMPOSIUM
  • Place of Presentation: コープイン京都（京都府）
  • Invited
• [Presentation] Molecular pathogenesis of Fanconi anemia (2012)
  • Author(s): Minoru Takata
  • Organizer: 第54回日本小児血液・がん学会学術集会
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Invited
• [Presentation] "Fanconi anemia and the DNA damage response" (2012)
  • Author(s): Minoru Takata
  • Organizer: The Sugahara Memorial International Symposium(招待講演)
  • Place of Presentation: 稲盛ホール（京都市）
• [Presentation] 「DNA損傷への応答メカニズムと疾患」 (2012)
  • Author(s): 高田穣
  • Organizer: 第11回分子予防環境医学研究会(招待講演)
  • Place of Presentation: 倉敷市市民会館（倉敷市）
• [Presentation] ファンコニ貧血コア複合体によるATR-ATRIPキナーゼのクロマチン動態制御 (2011)
  • Author(s): 高田穣、冨田純也、板谷(内田)亜希子、茂地智子、海野純也、前原善彦、石合正道
  • Organizer: 第70回日本癌学会学術総会 シンポジウム"Molecular basis of genome instability in cancer"
  • Place of Presentation: 名古屋市
• [Presentation] 「ファンコニ貧血とDNA損傷シグナリング」シンポジウム「International session for DNA repair and related subjects」 (2011)
  • Author(s): Takata,M
  • Organizer: 日 本放射線影響学会第54回大会 招待講演
  • Place of Presentation: 神戸市
• [Presentation] ファンコニ貧血コア複合体によるＡＴＲ－ＡＴＲＩＰキナーゼのクロマチン動態制御 (2011)
  • Author(s): 高田穣、冨田純也、板谷（内田）亜希子、茂地智子、海野純也、前原善彦、石合正道
  • Organizer: 第70回日本癌学会学術総会 シンポジウム"Molecular basis of genome instability in cancer"
  • Place of Presentation: 名古屋国際会議場（名古屋市）
• [Presentation] 「放射線被曝医療」司会 (2011)
  • Author(s): 高田穣
  • Organizer: 第28回日本医学会総会 講演会 特別企画プログラム(招待講演)
  • Place of Presentation: 東京ビッグサイト（東京都）
• [Presentation] An Absolute Requirement of ATRIP-ATR Kinase in Replication Stress-induced Triggering of the FA Pathway Activation (2011)
  • Author(s): T.Shigechi, J.Tomida, K.Sato, Y.Zhang, E.Uchida, J.Eykelenbo, N.Lowndes, H.Kurumizaka, Y.Maehara, M.Takata
  • Organizer: 23rd ANNUAL Fanconi Anemia Research Fund SCIENTIFIC SYMPOSIUM
  • Place of Presentation: Princesa Sofia Hotel（スペイン）
• [Presentation] International session for DNA repair and related subjects (2011)
  • Author(s): Takata, M
  • Organizer: 日本放射線影響学会第54回大会(招待講演)
  • Place of Presentation: 神戸商工会議所会館（神戸市）
• [Presentation] "Genome Dynamics: molecular network in maintenance and conversion" organizer (2011)
  • Author(s): 高田 穣
  • Organizer: 日本分子生物学会第34回年会(招待講演)
  • Place of Presentation: パシフィコ横浜（横浜市）
• [Presentation] Molecular mechanisms of the Fanconi anemia pathway (2011)
  • Author(s): Masamichi Ishiai, Junya Tomida, Tomoko Shigechi, Akiko Itaya, Emi Uchida, Junya Unno, Minoru Takata
  • Organizer: 日本分子生物学会第34回年会
  • Place of Presentation: パシフィコ横浜（横浜市）
• [Presentation] Fanconi anemia and the DNA damage signaling (2011)
  • Author(s): Minoru Takata
  • Organizer: DNA損傷応答シグナルにおけるクロマチン動態とエピジェネティクス制御, 27th RBC-NIRS International Symposium(招待講演)
  • Place of Presentation: コープイン京都（京都市）
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