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新たに見出された細胞周期関連遺伝子群の機能解析

Research Project

Project/Area Number 17770169
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Cell biology
Research InstitutionMeiji University

Principal Investigator

吉田 健一  明治大学, 農学部, 講師 (20345036)

Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2005: ¥2,700,000 (Direct Cost: ¥2,700,000)
Keywords分子生物学 / 転写因子 / 細胞周期 / 増殖・分化
Research Abstract

細胞周期関連遺伝子群(CDCA [cell-division-cycle-associated]1-5,7および8)として同定された、CDCA3のG2/M期特異的発現制御機構を、またCDCA7がc-Mycと共に転写因子E2F1により発現誘導される事実を解明した。以上の経緯より、他のCDCA遺伝子群の機能解析が、細胞周期のさらなる理解と精緻な分子機構解明に多大な貢献をし得ると実感し、本年度では、CDCA4の細胞内局在、過剰発現、RNA interference (RNAi)による発現抑制効果の検証、抗体作製などによる細胞周期における機能の解明を目指した。
CDCA4-GFPのHeLa細胞内局在から、CDCA4は核内に局在することが判明した。RNAiによるCDCA4発現抑制の結果、細胞の増殖性は亢進し、BrdU取り込みを指標としたDNA合成活性も顕著に増強した。一方、過剰発現細胞は得られなかったことから、CDCA4の発現充進が細胞の増殖に対して抑制的に作用している可能性が示唆された。CDCA4はE2F1の標的遺伝子として同定したが、興味深いことにE2F1の転写活性を抑制する転写コファクターである可能性が示唆された。CDCA4に対する抗体を作製する目的で、GST-CDCA4融合タンパクを大腸菌で産生させ、ポリクローナル抗体を作製した。本抗体により、内在性の33-kDaのCDCA4タンパクを検出し、細胞周期G1からS期にかけてタンパク発現の増加を見出した。CDCA4タンパクはHeLa以外でも、Jurkat、Daudi、293、Rh30、A375、T98G、HCT116およびHep2においても検出が確認された。今後、CDCA4遺伝子と細胞周期異常・細胞癌化との関連をさらに解明する必要がある。

Report

(2 results)
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • Research Products

    (12 results)

All 2006 2005

All Journal Article (12 results)

  • [Journal Article] Tumor necrosis factor-alpha stimulated gene-6 inhibits osteoblastic differentiation of human mesenchymal stem cells induced by OS and BMP-22006

    • Author(s)
      Tsukahara S, Ikeda R, Goto S, Yoshida K, Mitsumori R, Sakamoto Y, Tajima A, Yokoyama T, Toh S, Furukawa KI, Inoue I
    • Journal Title

      Biochemical Journal 398

      Pages: 595-603

    • Related Report
      2006 Annual Research Report
  • [Journal Article] The small heat shock protein alphaB-crystallin inhibits differentiation-induced caspase 3 activation and myogenic differentiation2006

    • Author(s)
      Ikeda R, Yoshida K, Ushiyama M, Yamaguchi T, Iwashita K, Futagawa T, Shibayama Y, Oiso S, Takeda Y, Kariyazono H, Furukawa T, Nakamura K, Akiyama S, Inoue I, Yamada K
    • Journal Title

      Biological & Pharmaceutical Bulletin 29

      Pages: 1815-1819

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Comparative genomics of MCM8 orthologous genes reveals transcriptional regulation by transcription factor E2F2006

    • Author(s)
      Hayashi R, Goto Y, Haga A, Kobayashi D, Ikeda R, Yoshida K
    • Journal Title

      Gene 367

      Pages: 126-134

    • Related Report
      2006 Annual Research Report
  • [Journal Article] JPO1/CDCA7, a novel transcription factor E2F1-induced protein, possesses intrinsic transcriptional regulator activity2006

    • Author(s)
      Goto Y, Hayashi R, Muramatsu T, Ogawa H, Eguchi I, Oshida Y, Ohtani K, Yoshida K
    • Journal Title

      Biochimica et Biophysica Acta 1759

      Pages: 60-68

    • Related Report
      2006 Annual Research Report
  • [Journal Article] A combined computational and experimental study on the structure-regulation relationships of putative mammalian DNA replication initiator GINS2006

    • Author(s)
      Hayashi R, Arauchi T, Tategu M, Goto Y, Yoshida K
    • Journal Title

      Genomics, Proteomics, & Bioinformatics 4

      Pages: 156-164

    • Related Report
      2006 Annual Research Report
  • [Journal Article] CDCA4 is an E2F transcription factor family-induced nuclear factor that regulates E2F-dependent transcriptional activation and cell proliferation2006

    • Author(s)
      Hayashi R, Goto Y, Ikeda R, Yokoyama KK, Yoshida K
    • Journal Title

      Journal of Biological Chemistry 281

      Pages: 35633-35648

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Comparative genomics of MCM8 orthologous genes reveals transcriptional regulation by transcription factor E2F2006

    • Author(s)
      Hayashi R
    • Journal Title

      Gene (in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Cell-cycle-dependent regulation of the human and mouse Tome-1 promoters2005

    • Author(s)
      Yoshida K
    • Journal Title

      FEBS Letters 579

      Pages: 1488-1492

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Identification of a novel cell-cycle-induced MCM family protein MCM92005

    • Author(s)
      Yoshida K
    • Journal Title

      Biochemical and Biophysical Research Communications 331

      Pages: 669-674

    • Related Report
      2005 Annual Research Report
  • [Journal Article] PLZF promotes osteoblastic differentiation of human mesenchymal stem cells as an upstream regulator of CBFA12005

    • Author(s)
      Ikeda R
    • Journal Title

      Journal of Biological Chemistry 280

      Pages: 8523-8530

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Identification and characterization of human ZNF18 gene in silico2005

    • Author(s)
      Yoshida K
    • Journal Title

      International Journal of Molecular Medicine 15

      Pages: 545-548

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Identification and characterization of a novel kelch-like gene KLHL15 in silico2005

    • Author(s)
      Yoshida K
    • Journal Title

      Ongology Reports 13

      Pages: 1133-1137

    • Related Report
      2005 Annual Research Report

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Published: 2005-04-01   Modified: 2016-04-21  

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