Molecular mechanism of genetic DNA rearrangements in fission yeast

• Project/Area Number: 21247027
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: IWASAKI Hiroshi 東京工業大学, 大学院・生命理工学研究科, 教授 (60232659)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥45,760,000 (Direct Cost: ¥35,200,000、Indirect Cost: ¥10,560,000); Fiscal Year 2011: ¥15,470,000 (Direct Cost: ¥11,900,000、Indirect Cost: ¥3,570,000); Fiscal Year 2010: ¥15,470,000 (Direct Cost: ¥11,900,000、Indirect Cost: ¥3,570,000); Fiscal Year 2009: ¥14,820,000 (Direct Cost: ¥11,400,000、Indirect Cost: ¥3,420,000)
• Keywords: 相同組換え / DNA二重鎖切断 / DNA修復 / DNA鎖交換反応 / 分裂酵母 / Rad22 / MRN複合体 / Ctp1 / Dmc1

Research Abstract

We are very much interested in molecular mechanism of genetic DNA rearrangements(e. g. homologous recombination), and thus during this proposed research period we tried to elucidate some functions of proteins involved in genetic rearrangements in fission yeast, which was used as a model biological system. Among several approaches we took for this aim, we had a great achievement for analysis of Dmc1 recombinase that works for meiotic recombination. We demonstrated fission yeast Dmc1-mediated Holliday junction formation in vitro. The directionality is 5'-3', which is the same as that by bacterial RecA but the opposite by Rad51 recombinases. Our finding is very important to understand for molecular mechanisms of meiotic recombination that accompanies high level of crossover products.

Research Products

• [Journal Article] Mechanistic insights into the activation of Rad51-mediated strand exchange from the structure of a recombination activator, the Swi5-Sfr1 complex (2012)
  • Author(s): Kuwabara N, Murayama Y, Hashimoto H, Kokabu Y, Ikeguchi M, Sato M, Mayanagi K, Tsutsui Y, Iwasaki H, Shimizu T
  • Journal Title: Structure Volume: 20 Pages: 440-449
  • Peer Reviewed
• [Journal Article] Opposing role of condensing hinge against replication protein A in mitosis and interphase through promoting DNA annealing (2011)
  • Author(s): Yuko Akai, Yumiko Kurokawa, Norihiko Nakazawa, Yuko Tonami-Murakami, Yuki Suzuki, Shige H.Yoshimura, Hiroshi Iwasaki, Yoshiharu Shiroiwa, Takahiro Nakamura, Eri Shibata, Mitsuhiro Yanagida
  • Journal Title: Open Biology Volume: 1 Issue: 4 Pages: 110023-110023
  • DOI: 10.1098/rsob.110023
  • Peer Reviewed
• [Journal Article] The fission yeast Swi5-Sfr1 complex, an activator of Rad51 recombinase, forms an extremely elongated Dogleg-shaped structure (2011)
  • Author(s): Kokabu Y, Murayama Y, Kuwabara N, Oroguchi T, Hashimoto H, Tsutsui Y, Nozaki N, Akashi S, Unzai S, Shimizu T, Iwasaki H, Sato M, Ikeguchi M
  • Journal Title: J Biol Chem Volume: 286 Pages: 43569-76
  • Peer Reviewed
• [Journal Article] Cells polarity in Saccharomyces cerevisiae depends on proper localization of the Bud9 landmark protein by the EKC/KEOPS complex (2011)
  • Author(s): Kato Y, Kawasaki H, Ohyama Y, Morishita T, Iwasaki H, Kokubo T, Hirano H
  • Journal Title: Genetics Volume: 188 Pages: 871-882
  • Peer Reviewed
• [Journal Article] An in vitro assay for monitoring the formation and branch migration of Holliday junctions mediated by a eukaryotic recombinase (2011)
  • Author(s): Murayama Y and Iwasaki H
  • Journal Title: Methods Mol. Biol Volume: 745 Pages: 385-405
  • Peer Reviewed
• [Journal Article] The fission yeast meiosis-specific Dmc1 recombinase mediates formation and branch migration of Holliday junctions by preferentially promoting strand-exchange in a direction opposite to that of Rad51 (2011)
  • Author(s): Murayama Y, Tsutui, Y and Iwasaki H
  • Journal Title: Genes Dev Volume: 25 Pages: 516-527
  • Peer Reviewed
• [Journal Article] The fission yeast Swi5-Sfr1 complex, an activator of Rad51 recombinase, forms an extremely elongated Dogleg-shaped structure (2011)
  • Author(s): Kokabu Y, Murayama Y, Kuwabara N, Oroguchi T, Hashimoto H, Tsutsui Y, Nozaki N, Akashi S, Unzai S, Shimizu T, Iwasaki H, Sato M, Ikeguchi M
  • Journal Title: J Biol Chem. Volume: 286 Pages: 43569-76
  • Peer Reviewed
• [Journal Article] An in vitro assay for monitoring the formation and branch migration of Holliday junctions mediated by a eukaryotic recombinase (2011)
  • Author(s): Murayama Y, Iwasaki H
  • Journal Title: Methods Mol.Biol. Volume: 745 Pages: 385-405
  • Peer Reviewed
• [Journal Article] The fission yeast meiosis-specific Dmcl recombinase mediates formation and branch migration of Holliday junctions by preferentially promoting strand-exchange in a direction opposite to that of Rad51. (2011)
  • Author(s): Murayama Y., Tsutui, Y., Iwasaki H.
  • Journal Title: Genes Dev. Volume: 25 Pages: 516-527
  • Peer Reviewed
• [Journal Article] Srs2 plays a critical role in reversible G2 arrest upon chronic and low doses of UV irradiation via two distinct homologous recombination-dependent mechanisms in post-replication repair deficient cells (2010)
  • Author(s): Hishida T, Hirade Y, Haruta N, Kubota Y and Iwasaki H
  • Journal Title: Mol. Cell. Biol Volume: 30 Pages: 4840-4850
  • Peer Reviewed
• [Journal Article] Purification and crystallization of Swi5 and Swi5-Sfr1 complex from fission yeast (2010)
  • Author(s): Kuwabara N, Hashimoto H, Yamada N, Unzai S, Ikeguchi M, Sato M, Murayama Y, Iwasaki H and Shimizu T
  • Journal Title: Acta Crystallogr Sect F Struct Biol Cryst Commun Volume: F66 Pages: 1124-1126
  • Peer Reviewed
• [Journal Article] Srs2 plays a critical role in reversible G2 arrest upon chronic and low doses of UV irradiation via two distinct homologous recombination-dependent mechanisms in post-replication repair deficient cells. (2010)
  • Author(s): Hishida T, Hirade Y, Haruta N, Kubota Y, Iwasaki H
  • Journal Title: Mol.Cell.Biol Volume: 30 Pages: 4840-4850
  • Peer Reviewed
• [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 and Tainer JA.
  • Journal Title: Cell Volume: 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, and Iwasaki H
  • Journal Title: Nature Volume: 457 Pages: 612-615
  • Peer Reviewed
• [Journal Article] Nbs 1 flexibly tethers Ctp 1 and Mre 11-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
• [Presentation] Regulation of Rad51 and Dmc1 recombinases in fission yeast (2011)
  • Author(s): Iwasaki, H
  • Organizer: New frontier of the research in Rad51 recombinase and its accessory proteins
  • Place of Presentation: Institut de Biologie Physico Chimique, Paris(招待講演)
  • Year and Date: 2011-09-28
• [Presentation] Reconstitution of two DNA strand exchange reactions during homologous recombination (2011)
  • Author(s): Iwasaki H.
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] Regulation of Rad51 recombinase in fission yeast. Japan-France conference on the RecA family recombinases : New Frontier of the research on Rad51 recombinases and its accessory proteins(Meeting organizer) (2011)
  • Author(s): Iwasaki H.
  • Organizer: Institut de Biologie Physico Chimique
  • Place of Presentation: Paris, France
• [Presentation] 相同組換えにおけるDNA鎖交換反応の制御機構 (2011)
  • Author(s): 岩崎博史
  • Organizer: 日本遺伝学会第83回大会
  • Place of Presentation: 京都大学農学部・農学研究科
• [Presentation] Differential effects of Rad22 on the Rad51-and Dmc1-mediated DNA strand exchange reactions (2011)
  • Author(s): Iwasaki H.
  • Organizer: 6^<th> international fission yeast meeting
  • Place of Presentation: Harvard Medical School, Boston, USA
• [Presentation] Molecular mechanism of DNA strand exchange reaction, a central reaction of homologous recombination mediated by recombinases (2011)
  • Author(s): Iwasaki H.
  • Organizer: China-Japan Symposium on Cancer Research
  • Place of Presentation: 深川,中国
• [Presentation] Molecular mechanism of DNA strand exchange reaction, a central reaction of homologous recombination mediated by recombinases (2011)
  • Author(s): Iwassaki, H
  • Organizer: China-Japan Symposium on Cancer Research
  • Place of Presentation: Shinzhen, China(招待講演)
• [Presentation] Differential effects of Rad22 on the Rad51- and Dmc1-mediated DNA strand exchange reactions (2011)
  • Author(s): Iwasaki, H
  • Organizer: 6^<th> international fission yeast meeting
  • Place of Presentation: Harvard Medical School, Boston, USA(招待講演)
• [Presentation] 組換え修復の分子機構 (2011)
  • Author(s): 岩崎博史
  • Organizer: 生化学若い研究者の会
  • Place of Presentation: 八王子セミナーハウス(招待講演)
• [Presentation] 相同組換えにおけるDNA鎖交換反応の制御機構 (2011)
  • Author(s): 岩崎博史
  • Organizer: 日本遺伝学会第83回大会
  • Place of Presentation: 京都大学農学部・農学研究科(招待講演)
• [Presentation] Reconstitution of two DNA strand exchange reactions during homologous recombination (2011)
  • Author(s): 岩崎博史
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(招待講演)
• [Presentation] 組換え修復とHolliday構造の分岐点移動反応の方向性 (2010)
  • Author(s): 岩崎博史
  • Organizer: 日本分子生物学会生化学会合同年会
  • Place of Presentation: 神戸ポートアイランド
• [Presentation] Biochemical differences between Rad51 and Dmc1 rcombinases from fission yeast (2010)
  • Author(s): Iwasaki H.
  • Organizer: 3R Symposium
  • Place of Presentation: 富山国際会議場
• [Presentation] 組換え修復とHolliday構造の分岐点移動反応の方向性 (2010)
  • Author(s): 岩崎博史
  • Organizer: 日本分子生物学会生化学会合同年会 BMB2010
  • Place of Presentation: 神戸ポートアイランド
• [Presentation] Characterization for in vitro sumoylation of Schizosaccharomyses pombe Rad22 protein involved in homologous recombination and recombination repair (2009)
  • Author(s): Yumiko Kurokawa, Yasuto Murayama, Yasuhiro Tsutsui, Fumiaki Yamao, Hiroshi Iwasaki
  • Organizer: 第32回 日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2009-12-10
• [Presentation] Biochemical analysis of fissionyeast recombinases and their auxiliaryproteins (2009)
  • Author(s): Iwasaki H.
  • Organizer: 5^<th> international fission yeast meeting
  • Place of Presentation: 国立オリンピック記念青少年総合センター、東京
• [Presentation] Regulation of strand exchange reactions by mediator proteins (2009)
  • Author(s): Iwasaki H.
  • Organizer: FASEB Summer Research Conference
  • Place of Presentation: Snowmass Village, Colorado, USA
• [Book] Human CtIP and its homolog : Team players in DSB resection games. In DNA repair. On the pathways to Fixing DNA Damage and Errors (2011)
  • Author(s): Tsutsui Y, Kawasaki A and Iwasaki H.
  • Total Pages: 380
  • Publisher: InTech社
• [Book] Human CtIP and its homologs : Team players in DSB resection games. in DNA Repair-On the pathways to Fixing DNA Damage and Errors(Ed Storici) (2011)
  • Author(s): Tsutsui Y, Kawasaki A, Iwasaki, H
  • Publisher: InTech
• [Remarks]
  • URL: http://www.iwasakilab.bio.titech.ac.jp/
• [Remarks]
  • URL: http://www.iwasakilab.bio.titech.ac.jp/