Functional Conversion and Artificial Repressor of Zinc Finger Proteins

锌指蛋白的功能转换和人工抑制

• 批准号: 09557200
• 负责人: SUGIURA Yukio
• 金额: $ 7.23万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557200/
• 关键词: Zinc finger; DNA binding; Base recognition; Artificial repressor; Transcription factor; Molecular design; Genomu information; Functional conversion; DNA認識

DNA recognition is a very important phenomenon in biology and chemistry, and the molecular basis of the sequence-specific DNA binding is the subject of investigations aimed at the rational design of artificial repressors. Zinc finger libraries with different DNA-binding specificites based on a one-finger-three-nuclestide code have been developed successfully. Newly designed zinc finger protein containing nine Cys_2His_2 type motifs were manipulated, and the DNA binding property of this multi finger protein was compared with that of nation three zinc finger protein Sp1. Evidently, the footprinting analysis demonstrated that the nine finger protein binds at least 27 contiguous base pairs of DNA sequence. Six zinc finger proteins induced sequence-specific DNA bending were also designed and a bent in the DNA was determined by electrophoretic methods. Such multi zinc finger protein may be useful as genome-specific transcriptional swiches in future.

DNA识别是生物学和化学中非常重要的现象，序列特异性DNA结合的分子基础是旨在合理设计人工阻遏物的研究主题。基于一指三核苷酸编码的具有不同DNA结合特异性的锌指文库已成功开发。对新设计的含有9个Cys_2His_2型基序的锌指蛋白进行了操作，并将这种多指蛋白的DNA结合特性与国三锌指蛋白Sp1进行了比较。显然，足迹分析表明九指蛋白结合了 DNA 序列的至少 27 个连续碱基对。还设计了六种锌指蛋白诱导的序列特异性 DNA 弯曲，并通过电泳方法测定了 DNA 的弯曲。这种多锌指蛋白将来可能可用作基因组特异性转录开关。

项目成果

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Kamiuchi, T.et al.：“具有 30 个碱基对结合位点的人工九锌指肽”生物化学。

Sugiura, Y.et al.: "Recognition of DNA se-quence and DNA bending. Molecular design for artificial repressors" J.Syn.Org.Chem.Jpn.55(5). 384-392 (1997)

Sugiura, Y.et al.：“DNA 序列和 DNA 弯曲的识别。人工阻遏物的分子设计”J.Syn.Org.Chem.Jpn.55(5)。

M.Yokono: "Unique DNA Binding Mode of the N-Terminal Zinc Finger of Transcription Factor Spl" Biochemistry. 37・19. 6824-6832 (1998)

M.Yokono：“转录因子Spl N末端锌指的独特DNA结合模式”生物化学37・19（1998）。

T.Kamiuti: "New Multi Zinc Finger Protein : Biosynthetic Design and Characteristics of DNA Recognition" Nucleic Acids Res.(Sym.Ser.). 37. 153-154 (1997)

T.Kamiuti：“新型多锌指蛋白：DNA 识别的生物合成设计和特征”核酸研究（Sym.Ser.）。

Yokono, M.et al.: "Unique DNA binding mode of the N-terminal zinc finger of transcription factor Sp1" Biochemistry. 37(19). 6824-6832 (1998)

Yokono, M.et al.：“转录因子 Sp1 N 末端锌指的独特 DNA 结合模式”生物化学。