Molecular mechanism of a recovery from stalled replication forks coupled with homologous recombination and its regulation

• Project/Area Number: 17080007
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Yokohama City University
• Principal Investigator: IWASAKI Hiroshi Yokohama City University, 大学院・生命理工学研究科, 教授 (60232659)
• Project Period (FY): 2005 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥63,500,000 (Direct Cost: ¥63,500,000); Fiscal Year 2009: ¥12,700,000 (Direct Cost: ¥12,700,000); Fiscal Year 2008: ¥12,700,000 (Direct Cost: ¥12,700,000); Fiscal Year 2007: ¥12,700,000 (Direct Cost: ¥12,700,000); Fiscal Year 2006: ¥12,700,000 (Direct Cost: ¥12,700,000); Fiscal Year 2005: ¥12,700,000 (Direct Cost: ¥12,700,000)
• Keywords: 相同組換え / 複製フォーク / チェックポイント / DNA二重鎖切断 / DNA修復 / リコンビナーゼ / Holliday構造 / 試験管内再構成系 / Dmc1 / 分裂酵母 / Rad51 / Rad22 / Rhp51 / Swi5-Sfr1 / Fbh1ヘリカーゼ / Rad51(Rhp51)リコンビナーゼ / Swi5-Sfr1複合体 / DNA鎖交換反応 / メディエーター / 接合型変換 / Rad55-Rad57(Rhp55-Rhp57)複合体

Research Abstract

Homologous recombination (HR) plays a crucial role in a recovery from stalled replication forks. Our long-term end is to elucidate molecular mechanism of a coupling reaction and a functional inter-play between DNA replication and HR. For this end, we cut into molecular mechanisms of key reactions for HR such as DNA strand exchange reaction and the Holliday intermediate formation. During the period of this study section, we established an in vitro reconstitution system of Swi5-Sfr1-Rad22-Rad51-dependent strand exchange reaction and Holliday junction formation. One of the most important findings is the 3' to 5' direction of formation and branch migration of Holliday junction.

Research Products

• [Journal Article] Nbs1 flexibly tethers Ctp1 and Mre11-Rad50 to coordinate DNA double-strand break processing and repair. (2009)
  • Author(s): Williams RS, Dodson GE, Limbo O, Yamada Y, Williams JS, Guenther G, Classen S, Glover JN, Iwasaki H, Russell P, Tainer JA.
  • Journal Title: Cell 139 Pages: 87-99
  • Peer Reviewed
• [Journal Article] RAD6-RAD18-RAD5 pathway-dependent tolerance to chronic low-dose UV light. (2009)
  • Author(s): Hishida T, Kubota Y, Carr AM, Iwasaki H.
  • Journal Title: Nature 457 Pages: 612-615
  • Peer Reviewed
• [Journal Article] RAD6-RAD18-RAD5 pathway-dependent tolerance to chronic low-dose UV light (2009)
  • Author(s): Hishida T, Kubota Y, Carr AM and Iwasaki H
  • Journal Title: Nature 457 Pages: 612-615
  • Peer Reviewed
• [Journal Article] A DNA polymeraseαaccessory protein, Mcl1, is required for propagation of centromere structures in fission yeast. (2008)
  • Author(s): Natsume T, Tsutsui Y, Toda T, Dunleavy EM, Pidoux AL, Iwasaki H, Shirahige K, Allshire RC, Yamao F.
  • Journal Title: PLoS One 3
  • Peer Reviewed
• [Journal Article] Molecular characterization of the Schizosaccharomyces pombe nip1+/ctp1+ gene in DNA double strand break repair in association with the Mre11-Rad50-Nbs1 complex. (2008)
  • Author(s): Akamatsu Y, Murayama Y, Yamada T, Nakazaki T, Tsutsui Y, Ohta K, Iwasaki H.
  • Journal Title: Mol. Cell. Biol. 28 Pages: 3639-3651
  • Peer Reviewed
• [Journal Article] Mus81 is essential for sister chromatid recombination at broken replication forks. (2008)
  • Author(s): Roseaulin L, Yamada Y, Tsutsui Y, Russell P, Iwasaki H, Arcangioli B.
  • Journal Title: EMBO J. 27 Pages: 1378-1387
  • Peer Reviewed
• [Journal Article] Reconstitution of DNA strand exchange mediated by Rhp51 recombinase and two mediators. (2008)
  • Author(s): Kurokawa Y, Murayama Y, Haruta- Takahashi N, Urabe I, Iwasaki H.
  • Journal Title: PLoS Biol. 6
  • Peer Reviewed
• [Journal Article] Formation and branch migration of Holliday junctions mediated by eukaryotic recombinases. (2008)
  • Author(s): Murayama Y, Kurokawa Y, Mayanagi K, Iwasaki H.
  • Journal Title: Nature 451 Pages: 1018-1021
  • Peer Reviewed
• [Journal Article] Fission yeast Swi5 protein, a novel DNA recombination mediator. (2008)
  • Author(s): Haruta N, Akamatsu Y, Tsutsui Y, Kurokawa Y, Murayama Y, Arcangioli B, Iwasaki H.
  • Journal Title: DNA Repair (Amst) 7 Pages: 1-9
  • Peer Reviewed
• [Journal Article] Reconstitution of DNA strand exchange mediated by Rhp51 recombinase and two mediators (2008)
  • Author(s): Kurokawa Y, Murayama Y, Haruta-Takahashi N, Urabe I, Iwasaki H
  • Journal Title: PLoS Biol 6
  • Peer Reviewed
• [Journal Article] Molecular characterization of the Schizosaccharomyces pombe nip1+/ ctp1+ gene in DNA double strand break repair in association with the Mre11-Rad50-Nbs1 complex (2008)
  • Author(s): Akamatsu Y, Murayama Y, Yama da T, Nakazaki T, Tsutsui Y, Ohta K, Iwasaki H
  • Journal Title: Mol Cell Biol 28 Pages: 3639-3651
  • Peer Reviewed
• [Journal Article] Reconstitution of DNA strand exchange mediated by Rhp51 recombinase and two mediators (2008)
  • Author(s): Kurokawa Y, Murayama Y, Haruta-Takahashi N, Urabe I, Iwasaki H.
  • Journal Title: PLoS Biol 6
  • Peer Reviewed
• [Journal Article] Molecular characterization of the Schizosaccharomyces pombe nip1+/ctp1+genein DNA double strand break repair in association with the Mre11-Rad50-Nbs1 complex (2008)
  • Author(s): Akamatsu Y, Murayama Y, Yamada T, Nakazaki T, Tsutsui Y, Ohta K, Iwasaki H.
  • Journal Title: Mol. Cell. Biol 28 Pages: 3639-3651
  • Peer Reviewed
• [Journal Article] Fission yeast Swi5/Sfr1 and Rhp55/Rhp57 differentially regulate Rhp51-dependent recombination outcomes. (2007)
  • Author(s): Akamatsu Y, Tsutsui Y, Morishita T, Siddique, MDP, Kurokawa, Y. Ikeguchi M, Yamao F, Arcangioli B, Iwasaki H.
  • Journal Title: EMBO J. 26 Pages: 1352-1362
  • Peer Reviewed
• [Journal Article] Fission yeast Swi5/Sfrl and Rhp55/Rhp57 differentially regulate Rhp51-dependent recombination outcomes. (2007)
  • Author(s): Akamatsu Y, Tsutsui Y, Morishita T, Siddique, MDP, Kurokawa, Y, Ikeguchi M, Fumiaki Yamao F, Arcangioli B, Iwasaki H
  • Journal Title: EMBO J. 26 Pages: 1352-1362
• [Journal Article] The Swi5-Sfr1 complex stimulates Rad51- and Dmc1-mediated DNA strand exchange in vitro. Nat. Struct. (2006)
  • Author(s): Haruta N, Kurokawa Y, Murayama Y, Akamatsu Y, Unzai S, Tsutsui Y, Iwasaki H.
  • Journal Title: Mol. Biol. 13 Pages: 823-730
  • Peer Reviewed
• [Journal Article] Direct observation of DNA rotation during branch migration of Holliday junction DNA by Escherichia coli RuvA-RuvB protein complex. (2006)
  • Author(s): Han YW, Tani T, Hayashi M, Hishida T, Iwasaki H, Shinagawa H, Harada Y.
  • Journal Title: Proc. Natl. Acad. Sci. USA. 103 Pages: 11544-11548
  • Peer Reviewed
• [Journal Article] The Swi5-Sfrl complex stimulates Rad51- and Dmcl-mediated DNA strand exchange in vitro (2006)
  • Author(s): Haruta N, Kurokawa Y, Murayama Y, Akamatsu Y, Unzai S, Tsutsui Y, Iwasaki H
  • Journal Title: Nat.Struct.Mol.Biol. 13 Pages: 823-730
• [Journal Article] Role of the Schizosaccharomyces pombe F-box DNA helicase in processing recombination intermediates. (2005)
  • Author(s): Morishita T, Furukawa F, Sakaguchi C, Toda T, Carr AM, Iwasaki H, Shinagawa H.
  • Journal Title: Mol. Cell. Biol. 25 Pages: 8074-8083
  • Peer Reviewed
• [Journal Article] Structure-function analysis of the three domains of RuvB motor protein. (2005)
  • Author(s): Ohnishi T, Hishida T, Harada Y, Iwasaki H, Shinagawa H.
  • Journal Title: J. Biol. Chem. 280 Pages: 30504-30510
  • Peer Reviewed
• [Journal Article] Genetic and physical interactions between Schizosaccharomyces pombe Mcl1 and Rad2, Dna2 and DNA polymerase alpha: evidence for a multifunctional role of Mcl1 in DNA replication and repair. (2005)
  • Author(s): Tsutsui Y, Morishita T, Natsume T, Yamashita K, Iwasaki H, Yamao F, Shinagawa H.
  • Journal Title: Curr. Genet. 48 Pages: 34-43
  • Peer Reviewed
• [Journal Article] Functional overlap between RecA and MgsA (RarA) in the rescue of stalled replication forks in Escherichia coli. (2005)
  • Author(s): Shibata T, Hishida T, Kubota Y, Han YW, Iwasaki H, Shinagawa H.
  • Journal Title: Genes Cells. 10 Pages: 181-191
  • Peer Reviewed
• [Presentation] BIOCHEMICAL ANALYSIS OF FISSION YEAST RECOMBINASES AND THEIR AUXILIARY PROTEINS (2009)
  • Author(s): Hiroshi Iwasaki
  • Organizer: The Fifth International Fission Yeast Meeting Pombe 2009
  • Place of Presentation: 国立オリンピック記念青少年総合センター
  • Year and Date: 2009-10-31
• [Presentation] Regulation of strand exchange reactions by mediator proteins (2009)
  • Author(s): Hiroshi Iwasaki
  • Organizer: FASEB SUMMER RESEARCH CONFERENCES
  • Place of Presentation: Snowmass Village, Colorado
  • Year and Date: 2009-08-04
• [Presentation] Biochemical analysis of fission yeast recombinaes and their auxiliary proteins. (2009)
  • Author(s): Iwasaki H.
  • Organizer: The Fifth International Fission Yeast Meeting (Pombe 2009).
  • Place of Presentation: Yoyogi Olympic Memorial Centre, Tokyo.
• [Presentation] Regulation of strand exchange reactions by mediator proteins. (2009)
  • Author(s): Iwasaki H.
  • Organizer: FASEB summer conference on " Genetic Recombination and Chromosome Rearrangements".
  • Place of Presentation: Snowmass, Colorado, USA.
• [Presentation] Biochemical analyses of the Swi5-Sfr1 complex on the two different recombainses in fission yeast. (2008)
  • Author(s): Iwasaki, H.
  • Organizer: The 6th international 3R symposium.
  • Place of Presentation: Tsumagoi Resort, Kakegawa, Shizuoka.
• [Presentation] 相同組換えにおけるDNA鎖交換の反応機構 (2008)
  • Author(s): 岩崎博史
  • Organizer: 第80回日本遺伝学会大会
  • Place of Presentation: 名古屋大学工学部
• [Presentation] DNA strand exchange reactions mediated by the fission yeast recombinase Rhp51 (spRad51) and its mediators, Rad22 (spRad52) and Swi5-Sfr1. (2008)
  • Author(s): Iwasaki, H.
  • Organizer: EMBO Conference Series on Recombination Mechanism.
  • Place of Presentation: Il Ciocco, Italy.
• [Presentation] The fission yeast nip1+ gene, which is involved in DNA double strand break repair togehter with the Mre11-Rad50-Nbs1 complex, is a functional counterpart Saccharomyces Sae2/Com1 and mammalian CtIP. (2008)
  • Author(s): Akamatsu Y, Yasuto Murayama Y, Yamada T, Tsutsui Y, Ohta K, Iwasaki H.
  • Organizer: DNA Replication and Recombination in Keystone Symposia.
  • Place of Presentation: Eldorado Hotel & Spa, Santa Fe, New Mexico, USA.
• [Presentation] Biochemical analysis of DNA strand exchange mediated by the fission yeast recombinase Rhp51 and its auxiliary proteins (2008)
  • Author(s): 岩崎博史
  • Organizer: 日本分子生物学会日本生化学会合同年会シンポジウム4S11染色体サイクルの制御機
  • Place of Presentation: 神戸ポートアイランド
• [Presentation] 分裂酵母Rhp51リコンビナーゼによるDNA鎖交換反応のメカニズム (2007)
  • Author(s): 岩崎博史
  • Organizer: 日本分子生物学会BMB2007
  • Place of Presentation: 横浜パシフィコ
  • Year and Date: 2007-12-14
• [Presentation] 分裂酵母Rhp51リコンビナーゼによるDNA鎖交換反応のメカニズム, 4S17シンポジ (2007)
  • Author(s): 岩崎博史
  • Organizer: 日本分子生物学会日本生化学会合同年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] Biochemical analysis of DNA strand exchange reactions mediated by fission yeast recombinase Rhp51. (2007)
  • Author(s): Iwasaki, H.
  • Organizer: The 2nd International Symposium on Chromosome Dynamics.
  • Place of Presentation: Senri-life science Center, Osaka.
• [Presentation] The Swi5-Sfr1 protein complex, a novel recombination mediator in fission yeast. (2007)
  • Author(s): Iwasaki H.
  • Organizer: The 4th International Fission Yeast Meeting (Pombe 2007)
  • Place of Presentation: Copenhagen, Denmark.
• [Presentation] Reconstitution of Rhp51 (spRad51)-dependent DNA strand exchange reaction during HR repair in fission yeast. (2007)
  • Author(s): Iwasaki, H.
  • Organizer: The 3rd US-Japan DNA repair meeting.
  • Place of Presentation: Akiu, Miyagi.
• [Presentation] 試験管内再構成系を用いた相同組換え反応の解析, 2006フォーラム. 分子生物学の未来 (2006)
  • Author(s): 岩崎博史
  • Organizer: 日本分子生物学会
  • Place of Presentation: 名古屋国際会議場
• [Presentation] Two parallel sub-pathways in Rad51-dependebt recombination repair in fission yeast. (2005)
  • Author(s): Iwasaki H.
  • Organizer: The 5th international 3R symposium.
  • Place of Presentation: Awaji, Hyogo, Japan.
• [Presentation] In vivo evidence that an S. pombe Swi5/Sfr1 complex is a novel Rad51 mediator. (2005)
  • Author(s): Iwasaki H.
  • Organizer: The51st NIBB Conference, New Aspects of Gene Amplification: Mechanisms and Biological Function.
  • Place of Presentation: Okazaki, Japan.
• [Presentation] Two Rad51-dependent recombination repair pathways in fission yeast. (2005)
  • Author(s): Iwasaki H.
  • Organizer: XXIIth International conference on yeast genetics and molecular biology.
  • Place of Presentation: Bratislava, Slovakia.
• [Book] 中低圧カラムクロマトグラフィー ～タンパク質の簡便な分離精製～. 実験医学別冊生命科学のための機器分析実験ハンドブック(西村善文編) (2007)
  • Author(s): 黒川裕美子, 岩崎博史
  • Publisher: 羊土社
• [Remarks]
  • URL: http://www.iwasakilab.bio.titech.ac.jp/
• [Remarks]
  • URL: http://www.iwasakilab.bio.titech.ac.jp/
• [Remarks]
  • URL: http://www.yokohama-cu.ac.jp/res/researcher/info/080207_iwasaki.html
• [Remarks]
  • URL: http://www.yokohama-cu.ac.jp/res/researcher/info/080207_iwasaki.html