Rad52のSUMO修飾による組換え制御機構の解析

2007 Fiscal Year Annual Research Report

• Project/Area Number: 19370073
• Research Institution: Tohoku University
• Principal Investigator: 関 政幸 Tohoku University, 大学院・薬学研究科, 准教授 (70202140)
• Keywords: 相同組換え / SUMO化 / Rad52 / チェックポイント / Rad51 / Ubc9 / 出芽酵母 / 核移行

Research Abstract

本年度の実績を下記に箇条書きする。
(1)DNA複製フォークにおけるRad52のSUMO修飾の役割の特定;複製チェックポイントが欠損したrad53やmec1変異株をMMS処理すると,複製フォークが崩壊する。この条件下、rad53やmec1変異株でRad52のSUMO化の異常な亢進が起こることを観察した。これはRad52の複製フォーク上での何らかの機能がrad53やmec1株で空回りしている可能性を示唆する。そこで,Rad52のSUMO修飾が起こらないrad52-KR変異株のMMS処理時の複製フォーク構造を2D-gel analysis法で観察したところrad52-K126Rやrad52-K200R変異株でフォーク上に生じるX-structureの形成阻害が観察され,それを報告した(DNA Repair 7,879-889,2008)。
(2)rad52-3KR変具によりMMS誘導性の相同組換えの効率が落ちる;Rad52のK10,K11,K220の3ヵ所全てをRに変換したrad52-3KR変異株ではRad52のSUMO化はほとんど起こらないが,この変異はMMS感受性などの表現型を示さない。次にMMS誘導性の相同組換えに影響が有無を調べたところ,世界ではじめてrad52-3KRにより相同組換えの効率が低下することを見いだし報告した(DNA Repair 7,879-889,2008)。
(3)Rad52は核移行後にSUMO化される;rad52-K200RにおいてもRad52のSUMO化が消失する。この原因がRad52の核移行がこの変異により低下することに起因することを立証し,それを報告した(Biochem.Biophys.Res.Commun.372,126-130,2008)。

Research Products

• [Journal Article] Nuclear localization of Rad52 is pre-requisite for its sumoylation (2008)
  • Author(s): Ohuchi T, Seki M, Enomoto T.
  • Journal Title: Biochem. Biophys. Res. Commun. 372 Pages: 126-130
  • Peer Reviewed
• [Journal Article] Rad52 sumoylation and its involvement in the efficient induction of homologous recombination (2008)
  • Author(s): Ohuchi T, Seki M, Branzei D, Maeda D, Ui A, Ogiwara H, Tada S, Enomoto T.
  • Journal Title: DNA Repair 7 Pages: 879-889
  • Peer Reviewed
• [Journal Article] Analyses of functional interaction between RECQL1, RECQL5, and BLM which physically interact with DNA topoisomerase IIIa (2008)
  • Author(s): Otsuki M, Seki M, Inoue E, Abe T, Narita Y, Yoshimura A, Tada S, Ishii Y, Enomoto T
  • Journal Title: Biochim. Biophys. Acta. 1782 Pages: 75-81
  • Peer Reviewed
• [Journal Article] Functional interactions between BLM and XRCC3 in the cell (2007)
  • Author(s): Otsuki M, Seki M, Inoue E, Yoshimura A, Kato G, Yamanouchi S, Kawabe Y, Tada S, Shinohara A, Komura J, Ono T, Takeda S, Ishii Y, Enomoto T.
  • Journal Title: J. Cell Biol. 179 Pages: 53-63
  • Peer Reviewed
• [Journal Article] Ctf18 is required for homologous recombination-mediated double-strand break repair (2007)
  • Author(s): Ogiwara. H., Ohuchi, T, Ui, A., Tada, S., Enomoto, T., Seki, M.
  • Journal Title: Nucleic Acids Res. 35 Pages: 4989-5000
  • Peer Reviewed
• [Journal Article] Rmi1, a member of the Sgs1-Top3 complex in budding yeast, contributesto sister chromatid cohesion (2007)
  • Author(s): Lai, M. S., Seki. M., Ui, A., Enomoto, T.
  • Journal Title: EMBO Rep. 8 Pages: 685-690
  • Peer Reviewed
• [Journal Article] Activation of a novel pathway involving Mms1 and Rad59 in sgs1 cells (2007)
  • Author(s): Ui A, Seki M, Ogiwara H, Lai MS, Yamamoto K, Tada S, Enomoto T.
  • Journal Title: Biochem. Biophys. Res. Commun. 356 Pages: 1031-1037
  • Peer Reviewed
• [Journal Article] Possible involvement of RecQL4 in the repair of double-strand DNA breaks in Xenopus egg extracts (2007)
  • Author(s): Kumata Y, Tada S, Yamanada Y, Tsuyama T, Kobayashi T, Dong YP, Ikegami K, Murofushi H, Seki M, Enomoto T.
  • Journal Title: Biochim. Biophys. Acta. 1773 Pages: 556-564
  • Peer Reviewed
• [Journal Article] WRN counteracts the NHEJ pathway upon camptothecin exposure (2007)
  • Author(s): Otsuki M, Seki M, Kawabe Y, Inoue E, Dong YP, Abe T, Kato G, Yoshimura A, Tada S, Enomoto T.
  • Journal Title: Biochem. Biophys. Res. Commun. 355 Pages: 477-482
  • Peer Reviewed
• [Journal Article] WRN functions in a RAD18-dependent damage avoidance pathway (2007)
  • Author(s): Dong YP, Seki M, Yoshimura A, Inoue E, Furukawa S, Tada S, Enomoto T
  • Journal Title: Biol. Pharm. Bull. 30 Pages: 1080-1083
  • Peer Reviewed
• [Journal Article] Rad50 is involved in MMS-induced recombination between homologous chromosomes in mitotic cells (2007)
  • Author(s): Tomizawa Y, Ui A, Onoda F, Ogiwara H, Tada S, Enomoto T, Seki M.
  • Journal Title: Genes Genet. Syst. 82 Pages: 157-160
  • Peer Reviewed
• [Journal Article] The INO80 chromatin remodeling complex functions in sister chromatid cohesion (2007)
  • Author(s): Ogiwara H, Enomoto T, Seki M.
  • Journal Title: Cell Cycle 6 Pages: 1090-1095
  • Peer Reviewed
• [Journal Article] Actin-related protein Arp4 functions in kinetochore assembly (2007)
  • Author(s): Ogiwara H, Ui A, Kawashima S, Kugou K, Onoda F, Iwahashi H, Harata M, Ohta K, Enomoto T, Seki M.
  • Journal Title: Nucleic Acids Res. 35 Pages: 3109-3117
  • Peer Reviewed
• [Presentation] Requirement of specific amino acids in Rad52 for sumoylation and recombination (2007)
  • Author(s): 関政幸
  • Organizer: 第三回 Japan-US DNA Repair Meeting
  • Place of Presentation: 仙台市/秋保/ホテルク レセント
  • Year and Date: 2007-05-09