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Roles of chromatin structure in gene conversion at chicken immunoglobulin locus

Research Project

Project/Area Number 14580689
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Molecular biology
Research InstitutionRIKEN Discovery Research Institute

Principal Investigator

OHTA Kunihiro  RIKEN Discovery Research Institute, Genetic Dynamics Research Unit, Unit leader, 遺伝ダイナミクス研究ユニット, 研究ユニットリーダー (90211789)

Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywordsrecombination / transcription / DT40 / chromatin / histone / acetylation / antibody
Research Abstract

In the chicken B-cell line DT40, gene conversion, a type of homologous recombination, primarily contributes to diversification of the immunoglobulin (Ig) gene. We have studied roles of chromatin structure in the regulation of this gene conversion in the chicken DT40 cell lines. We found that recombination-active Ig locus exhibits much higher levels of histone acetylation. Treatment of DT40 cells with histone deacetylase inhibitor Tricostatin A (TSA) markedly induced gene conversion frequency and transcriptional activity at recombination-active allele, but observed elevation of histone acetylation levels in both recombination-active and inactive-alleles, suggesting links between recombination activation and transcriotional activity.
Using DT40 cells harboring an artificial chromosomal recombination marker including partial GFP sequence, complete CFP sequence combined with inducible Tet promoter, 3' enhancer, and a matrix attachment region sequence, we further examined relationship between transcriptional activity and gene conversion frequency. After transcriptional activation, gene conversion events between partial GFP and CFP sequences were induced; thereby generating GFP-type fluorescence was detected in a small portion of the DT40 cells. Thus, it is suggested that gene conversion may be activated coupled to some transcription-related mechanisms.
In addition, using the effects by TSA on gene conversion, we developed a rapid and flexible system to produce specific monoclonal antibodies using the DT40 cells undergoing activated gene conversion. The selection system (ADLib: Autonomously Diversifying Library) enables quick establishment (〜one week from a diversifying library) of various clones producing monoclonal IgMs with enough specificity and affinity for immunological assays, and is applicable to various biotechnologies including rational protein design.

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (24 results)

All 2005 2004 2003 Other

All Journal Article (11 results) Book (1 results) Patent(Industrial Property Rights) (2 results) Publications (10 results)

  • [Journal Article] Rapid generation of specific antibodies by enhanced homologous recombination2005

    • Author(s)
      Seo H.et al.
    • Journal Title

      Nature Biotechnology 23

      Pages: 731-735

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Roles of histone acetylation and chromatin ermodeling factor in a meiotic recombination2004

    • Author(s)
      Ymada T. et al.
    • Journal Title

      EMBO J. 23

      Pages: 1792-1803

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Fission yeast global repressors regulate the specificity of chromatin alteration in response to distinct environment stresses2004

    • Author(s)
      Hirota K. et al.
    • Journal Title

      Nucleic Acids Research 32

      Pages: 855-862

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Roles of histone acetylation and chromatin remodeling factor in a meiotic recombination hotspot2004

    • Author(s)
      Yamada T. et al.
    • Journal Title

      EMBO J 23

      Pages: 1972-1803

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Fission yeast global repressors regulate the specificity of chromatin alteration in response to distinct environmental stresses2004

    • Author(s)
      Hirota K. et al.
    • Journal Title

      Nucleic Acids Research 32

      Pages: 855-862

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Fission yeast Tup1-like repressors repress chromatin remodeling at the fbp1^+ promoter and the ade6-M26 recombination hotspot2003

    • Author(s)
      Hirota K. et al.
    • Journal Title

      Genetics 165

      Pages: 505-515

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Gef1p and Scd1p, the two GDP-GTP exchange factors for Cdc42p, from a ring structure that shrinks during cytokinesis in Schizosaccharomyces prombe2003

    • Author(s)
      Hirota K. et al.
    • Journal Title

      Mol.Biol.Cell 14

      Pages: 3617-3627

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Correlation between premeiotic DNA replication and chromatin transition at yeast recombination inititation sites2003

    • Author(s)
      Murakami H. et al.
    • Journal Title

      Nucleic Acids Research 31

      Pages: 4085-4090

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Gef1 p and Scd1 p, the two GDP-GTP exchange factors for Cdc42p, form a ring structure that shrinks during cytokinesis in Schizosaccharomyces pombe2003

    • Author(s)
      Hirota K. et al.
    • Journal Title

      Mol. Biol. Cell 14

      Pages: 3617-3627

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Correlation between premeiotic DNA replication and chromatin transition at yeast recombination initiation sites2003

    • Author(s)
      Murakami H. et al.
    • Journal Title

      Nucleic Acids Research 31

      Pages: 4085-4090

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Rapid generation of specific antibodies by enhanced homologous recombination

    • Author(s)
      Seo H.et al.
    • Journal Title

      Nature Biotech. (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Book] クロマチンと遺伝子機能制御(堀越正美編):相同組換えとクロマチン構造2003

    • Author(s)
      太田邦史
    • Total Pages
      270
    • Publisher
      シュプリンガーフェアラーク東京
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Patent(Industrial Property Rights)] 体細胞相同組換えの促進方法及び特異的抗体の作製方法2003

    • Inventor(s)
      太田邦史, 瀬尾秀宗, 柴田武彦
    • Industrial Property Rights Holder
      (独)理化学研究所, (独)科学技術振興機構
    • Filing Date
      2003-07-28
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Patent(Industrial Property Rights)] 体細胞相同組換えの誘発方法2003

    • Inventor(s)
      太田邦史, 瀬尾秀宗, 柴田武彦
    • Industrial Property Rights Holder
      (独)理化学研究所, (独)科学技術振興機構
    • Filing Date
      2003-12-22
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yamada T. et al.: "Roles of histone acetylation and chromatin remodeling factor in a meiotic recombination hotspot"EMBO J.. 印刷中. (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hirota K. et al.: "Fission yeast global repressors regulate the specificity of chromatin alteration in response to distinct environmental stresses"Nucleic Acids Research. 32. 855-862 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hirota K. et al.: "Fission yeast Tup1-like repressors repress chromatin remodeling at the fbp1^+ promoter and the ade6-M26 recombination hotspot"Genetics. 165. 505-515 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hirota K. et al.: "Gef1p and Scd1p, the two GDP-GTP exchange factors for Cdc42p, form a ring structure that shrinks during cytokinesis in Schizosaccharomyces pombe"Mol.Biol.Cell. 14. 3617-3627 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tomita K. et al.: "Competition between the Rad50 complex and the Ku heterodimer reveals a role for Exo1 in processing double-strand break, but not telomeres"Mol.Cell.Biol. 23. 5186-5197 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Murakami H. et al.: "Correlation between premeiotic DNA replication and chromatin transition at yeast recombination initiation sites"Nucleic Acids Research. 31. 4085-4090 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 太田邦史: "クロマチンと遺伝子機能制御(堀越正美編)における「相同組換えとクロマチン構造」"シュプリンガーフェアラーク東京. 270 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Pecina A, Smith KN, Mezard C, Murakami H, Ohta K, Nicolas A: "Target stimulation of mejotic recombination"Cell. 111. 173-184 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Marin A., Gallardo M., Kato Y., Shirahige K., Gutierrez G., Ohta K., Aguilera A.: "Relationship between G+C content, ORF-length and mRNA concentration in Saccharomyces cerevisiae"Yeast. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Farah JA, Hartsuiker E, Mizuno K, Ohta K, Smith GR: "A 160-bp palindrome is a Rad50.Rad32-dependent mitotic recombination hotspot in Schizosaccharomyces pombe"Genetics. 161. 461-468 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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