• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Regulation of cellular genetic function by new DNA bending fingers

Research Project

Project/Area Number 13557210
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 医薬分子機能学
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

SUGIURA Yukio  Kyoto Univ., Inst.Chem.Res., Prof., 化学研究所, 教授 (40025698)

Co-Investigator(Kenkyū-buntansha) NAGAOKA Makoto  Kyoto Univ., Inst.Chem.Res., Assistant Prof., 化学研究所, 助手 (60314275)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥14,000,000 (Direct Cost: ¥14,000,000)
Fiscal Year 2003: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2001: ¥6,500,000 (Direct Cost: ¥6,500,000)
KeywordsZinc finger / Artificial protein / DNA recognition / DNA motif / DNA bending / Gene target / Artificial restriction enzyme / Artificial repressor / マルチフィンガー / 遺伝子制御 / 転写調節 / アーキテクチャー / リンカー / 転写因子 / DNA結合
Research Abstract

DNA bending is very important for various biological reactions. In particular, many transcription factors have been known to induce DNA bending and to support formation of the transcriptional initiation complex. We crated new artificial 6-zinc finger peptides that effectively induced DNA bending. The two DNA binding domains of transcription factor Sp1 were connected through polyglycine linker. The obtained 6-zinc finger proteins Sp1ZF6(Gly)n (n=4, 7, 10) were compared with native 3-zinc finger peptide Sp1 in the respect of DNA bending ability. Gel electrophoretic methods revealed that Sp1ZF6(Gly)_7 and Sp1ZF6(Gly)_<10> bound to two distal GC boxes and resulted in DNA bending. The phasing assays strongly suggested that the induced DNA bending was directed toward the major groove and that Sp1ZF6(Gly)_7 caused the most drastic directional change in DNA bending. Of special interest is the fact that the linker length between two 3-zinc finger motifs has a crucial effect on the entire DNA bending direction. In addition, the flexible and neutral linker of Sp1ZF6(Gly)_<10> was also converted to charged linkers including arginine and glutamate residues. In order to clarify the relation between the DNA bending ability and the transcriptional activity, we also reporter gene assays. The experimental results led to the activation of gene expression by the DNA bending due to artificial zinc finger peptide. Such DNA bending fingers may be feasible for use as a gene expression regulator based on the structural change in DNA in the near future.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] M.Imanishi: "Artificial DNA-Bending Six-Zinc Finger Peptides with Different Charged Linkers : Distinct Kinetic Properties of DNA Bindings"Biochemistry. 41・4. 1328-1334 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Nagaoka: "Novel Strategy for the Design of a New Zinc Finger : Creation of a Zinc Finger for the AT-Rich Sequence by α-Helix Substitution"J.Am.Chem.Soc.. 124・23. 6526-6527 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Nagaoka: "Interconversion between Serine and Aspartic Acid in the α Helix of the N-Terminal Zinc Finger of Sp1 : Implication for General Recognition Code and for Design of Novel Zinc Finger Peptide Recognizing Complementary Strand"Biochemistry. 41・28. 8819-8825 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] A.Nomura: "Contribution of Individual Zinc Ligands to Metal Binding and Peptide Folding of Zinc Finger Peptides"Inorg.Chem.. 41・14. 3693-3698 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Nagaoka: "Influence of amino acid numbers between two ligand cysteines of zinc finger proteins"Biochem.Biophys.Res.Commun.. 296・3. 553-559 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] W.Nomura: "Effects of Length and Position of an Extended Linker on Sequence-Selective DNA Recognition of Zinc Finger Peptides"Biochemistry. 42・50. 14805-14813 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Imanishi, M.et al.: "Artificial DNA-Bending Six-Zinc Finger Peptides with Different Charged Linkers : Distinct Kinetic Properties of DNA Bindings"Biochemistry. 41(4). 1328-1334 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nagaoka, M.et al.: "Novel Strategy for the Design of a New Zinc Finger : Creation of a Zinc Finger for the AT Rich Sequence by α-Helix Substitution"J.Am.Chem.Soc.. 124(23). 6526-6527 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nagaoka, M.et al.: "Interconversion between Serine and Aspartic Acid in the α Helix of the N-Terminal Zinc Finger of Sp1 : Implication for General Recognition Code and for Design of Novel Zinc Finger Peptide Recognizing Complementary Strand"Biochemistry. 41(28). 8819-8825 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nomura, A.et al.: "Contribution of Individual Zinc Ligands to Metal Binding and Peptide Folding of Zinc Finger Peptides"Inorg.Chem.. 41(14). 3693-3698 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nagaoka, M.et al.: "Influence of Amino Acid Numbers Between Two Ligand Cystenes of Zinc Finger Proteins"Biochem.Biophys.Res.Commun.. 296(3). 553-559 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nomura, W.et al.: "Effects of Length and Position of an Extended Linker on Sequence-Selective DNA Recognition of Zinc Finger Peptides"Biochemistry. 42(50). 14805-14813 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Matsushita: "GFP-Linked Zinc Finger Protein Sp1 : Fluorescence Study and Implication for N-Terminal Zinc Finger 1 as Hinge Finger"Bioorg.Med.Chem.. 11・1. 53-58 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] W.Nomura: "Influence of TFIIIA-Type Linker at the N- or C-Terminal of Nine-Zinc Finger Protein on DNA-Binding Site"Biochem.Biophys.Res.Commun.. 300・1. 87-92 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] W.Nomura: "Effects of Length and Position of an Extended Linker on Sequence-Selective DNA Recognition of Zinc Finger Peptides"Biochemistry. 42・50. 14805-14813 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] M.Imanishi: "Artificial DNA-Bending Six-Zinc Finger Peptides with Different Charged Linkers: Distinct Kinetic Properties of DNA Bindings"Biochemistry. 41・4. 1328-1334 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Nagaoka: "Interconversion between Serine and Aspartic Acid in the α Helix of the N-Terminal Zinc Finger of Sp1 : Implication for General Recognition Code and for Design of Novel Zinc Finger Peptide Recognizing Complementary Strand"Biochemistiy. 41・28. 8819-8825 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Nagaoka: "Novel Strategy for the Design of a New Zinc Finger : Creation of a Zinc Finger for the AT-Rich Sequence by α-Helix Substitution"J.Am.Chem.Soc.. 124・23. 6526-6527 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Nagaoka: "Multiconnection of identical zinc finger : Implication for DNA binding affinity and unit modulation of the three zinc finger domain"Biochemistry. 40・9. 2932-2941 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] M.Nagaoka: "Significant effect of linker sequence on DNA recognition by multi-zinc finger protein"Biochem.Biophys.Res.Commun.. 282・4. 1001-1007 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] M.Nagaoka: "Selected base sequence outside the target binding site of zinc finger protein Sp1"Nucleic Acids Res.. 29・24. 4920-4929 (2001)

    • Related Report
      2001 Annual Research Report

URL: 

Published: 2001-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi