Structural dynamics and functions of proteins involved in homologous recombination

2006 Fiscal Year Final Research Report Summary

• Project/Area Number: 17570097
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Waseda University
• Principal Investigator: KURUMIZAKA Hitoshi Faculty of Science and Engineering, Associate, Professor, 理工学術院, 助教授 (80300870)
• Project Period (FY): 2005 – 2006
• Keywords: homologous recombination / double strand break repair / genetic recombination / Rad51 / Dmc1 / meiosis / RecA / ATPase

Research Abstract

Chromosomal DNA is exposed to various DNA-damaging agents and sustains damage that induces genomic instability. A double-strand break (DSB) is caused by ionizing radiation, crosslinking reagents, oxidative stress, and DNA replication failure. If the DSB is left unrepaired, then cell death occurs. Homologous recombination is one of the major DSB repair pathways. This repair pathway is essentially error-free, since a homologous region of the undamaged sister chromatid is used as the template for repair. In contrast to the mitotic DSB repair pathway, meiotic cell division involves homologous recombination between homologous chromosomes, but not between sister chromatids. This preferential recombination between homologous chromosomes is initiated by the formation of a programmed double strand break (DSB), and ensures correct chromosomal segregation at meiosis I through the formation of chiasmata, which physically connect homologous chromosomes. Thus, homologous recombination is important to maintain the integrity of the chromosome in both mitotic and meiotic cells.
In homologous recombination, a single-stranded DNA (ssDNA) tail, produced at the DSB site, is incorporated into a nucleoprotein complex called the presynaptic filament. This presynaptic filament catalyzes homologous pairing and strand exchange with an intact homologous region of the double-stranded DNA (dsDNA) molecule. The bacterial RecA protein is known to form helical presynaptic filaments and to play central roles in homologous recombination. In eukaryotes, two homologs of RecA, the Rad51 and Dmcl proteins, which are conserved from yeast to human, are assumed to fulfill this role.
We have determined the crystal structure of the human Dmcl protein as an octameric ring form. Then, we performed the structure-based mutation analyses of the Rad51 and Dmcl proteins, and identified residues that function in DNA-binding and octameric ring formation.

Research Products

• [Journal Article] Genetic variance modifies apoptosis susceptibility in mature oocytes via alterations in DNA repair capacity and mitochondrial ultrastructure (2007)
  • Author(s): Perez GI, Acton BM, Jurisicova A, Perkins GA, White A, Brown J, Trbovich AM, Kim MR, Fissore R, Xu J, Ahmady A, D'Estaing SG, Li H, Kagawa W, Kurumizaka H, Yokoyama S, Okada H, Mak TW, Ellisman MH, Casper RF, Tilly JL
  • Journal Title: Cell Death Differ. 14(3) Pages: 524-533
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] HIV-1 Vpr induces ATM-dependent cellular signal with enhanced homologous recombination (2007)
  • Author(s): Nakai-Murakami C, Shimura M, Kinomoto M, Takizawa Y, Tokunaga K, Taguchi T, Hoshino S, Miyagawa K, Sata T, Kurumizaka H, Yuo A, Ishizaka Y
  • Journal Title: Oncogene 26(4) Pages: 477-486
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Genetic variance modifies apoptosis susceptibility in mature oocytes via alterations in DNA repair capacity and mitochondrial ultrastructure. (2007)
  • Author(s): Perez GI, Acton BM, Jurisicova A, Perkins GA, White A, Brown J, Trbovich AM, Kim MR, Fissore R, Xu J, Ahmady A, D'Estaing SG, Li H, Kagawa W, Kurumizaka H, Yokoyama S, Okada H, Mak TW, Ellisman MH, Casper RF, Tilly JL.
  • Journal Title: Cell Death Differ. 14(3) Pages: 524-533
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] HIV-1 Vpr induces ATM-dependent cellular signal with enhanced homologous recombination. (2007)
  • Author(s): Nakai-Murakami C, Shimura M, Kinomoto M, Takizawa Y, Tokunaga K, Taguchi T, Hoshino S, Miyagawa K, Sata T, Kurumizaka H, Yuo A, Ishizaka Y.
  • Journal Title: Oncogene 26(4) Pages: 477-486
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Stimulation of Dmcl-mediated DNA strand exchange by the human Rad54B protein (2006)
  • Author(s): Sarai N, Kagawa W, Kinebuchi T, Kagawa A, Tanaka K, Miyagawa K, Ikawa S, Shibata T, Kurumizaka H, Yokoyama S
  • Journal Title: Nucleic Acids Res. 34(16) Pages: 4429-4437
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Roles of the human Rad51 L1 and L2 loops in DNA binding (2006)
  • Author(s): Matsuo Y, Sakane I, Takizawa Y, Takahashi M, Kurumizaka H
  • Journal Title: FEBS Journal 273(14) Pages: 3148-3159
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The DnaA homolog of the hyperthermophilic eubacterium Thermotoga maritima forms an open complex with a minimal 149-bp origin region in an ATP-dependent manner (2006)
  • Author(s): Ozaki S, Fujimitsu K, Kurumizaka H, Katayama T
  • Journal Title: Genes Cells 11(4) Pages: 425-438
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Stimulation of Dmc1-mediated DNA strand exchange by the human Rad54B protein. (2006)
  • Author(s): Sarai N, Kagawa W, Kinebuchi T, Kagawa A, Tanaka K, Miyagawa K, Ikawa S, Shibata T, Kurumizaka H, Yokoyama S.
  • Journal Title: Nucleic Acids Res. 34(16) Pages: 4429-4437
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Roles of the human Rad51 L1 and L2 loops in DNA binding. (2006)
  • Author(s): Matsuo Y, Sakane I, Takizawa Y, Takahashi M, Kurumizaka H.
  • Journal Title: FEBS J. 273(14) Pages: 3148-3159
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The DnaA homolog of the hyperthermophilic eubacterium Thermotoga maritima forms an open complex with a minimal 149-bp origin region in an ATP-dependent manner. (2006)
  • Author(s): Ozaki S, Fujimitsu K, Kurumizaka H, Katayama T.
  • Journal Title: Genes Cells 11(4) Pages: 425-438
  • Description: 「研究成果報告書概要(欧文)」より