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，并且序列特异性DNA结合的分子基础是旨在合理设计人工阻遏物的研究的主题。基于一指三核苷酸编码的锌指文库已被成功构建。对新设计的含9个Cys_2His_2型基序的锌指蛋白进行了操作，并与国内三种锌指蛋白Sp1的DNA结合特性进行了比较。显然，足迹分析表明，九指蛋白结合至少27个连续碱基对的DNA序列。还设计了六种锌指蛋白诱导的序列特异性DNA弯曲，并通过电泳方法确定了DNA中的弯曲。这种多锌指蛋白有可能成为基因组特异性转录开关。

项目成果

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杉浦 幸雄: "DNA塩基配列およびDNA湾曲の認識" 有機合成化学協会誌. 55. 384-392 (1997)

Yukio Sugiura：“DNA 碱基序列和 DNA 曲率的识别”有机合成化学学会杂志 55. 384-392 (1997)。

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)。

Kamiuchi, T.et al.: "Artificial nine zinc finger peptide with 30 base pair binding sites" Biochemistry. 37(39). 13827-13834 (1998)

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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）。