Pomerantz J L, Sharp P A, Pabo C O
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Science. 1995 Jan 6;267(5194):93-6. doi: 10.1126/science.7809612.
Computer modeling suggested that transcription factors with novel sequence specificities could be designed by combining known DNA binding domains. This structure-based strategy was tested by construction of a fusion protein, ZFHD1, that contained zinc fingers 1 and 2 from Zif268, a short polypeptide linker, and the homeodomain from Oct-1. The fusion protein bound optimally to a sequence containing adjacent homeodomain (TAATTA) and zinc finger (NGGGNG) subsites. When fused to an activation domain, ZFHD1 regulated promoter activity in vivo in a sequence-specific manner. Analysis of known protein-DNA complexes suggests that many other DNA binding proteins could be designed in a similar fashion.
计算机建模表明，通过组合已知的DNA结合结构域，可以设计出具有新序列特异性的转录因子。通过构建融合蛋白ZFHD1对这种基于结构的策略进行了测试，该融合蛋白包含来自Zif268的锌指1和2、一个短多肽接头以及来自Oct-1的同源结构域。该融合蛋白与包含相邻同源结构域（TAATTA）和锌指（NGGGNG）亚位点的序列具有最佳结合。当与激活结构域融合时，ZFHD1在体内以序列特异性方式调节启动子活性。对已知蛋白质-DNA复合物的分析表明，许多其他DNA结合蛋白也可以以类似的方式设计。
