Hirata Tsuyoshi, Nomura Wataru, Imanishi Miki, Sugiura Yukio
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
Bioorg Med Chem Lett. 2005 May 2;15(9):2197-201. doi: 10.1016/j.bmcl.2005.03.040.
To assess the possibility of multi-connection of zinc finger domains for understanding of DNA binding mechanisms and gene regulation, the longest artificial zinc finger protein, Sp1ZF15, has been constructed. This zinc finger consists of 5 units of Sp1 zinc finger peptide connected by canonical short linker sequences (TGEKP). Recognition of the 50 base pairs of DNA and potential binding to shorter targets by Sp1ZF15 were determined. Sequence alterations of the GCG triplet to ATA at a target site clearly showed that Sp1ZF15 changes its DNA binding mode depending on the target sequences. Of special interest is the fact that Sp1ZF15 controls the number of finger domains active in DNA binding corresponding to the length and sequence of the target DNA. These results suggest that artificial transcription factors based upon these multi-zinc finger proteins have great potential for the regulation of a vast number of cellular processes.
为了评估锌指结构域多连接对于理解DNA结合机制和基因调控的可能性，构建了最长的人工锌指蛋白Sp1ZF15。该锌指由5个Sp1锌指肽单元通过典型的短连接序列（TGEKP）连接而成。确定了Sp1ZF15对50个碱基对DNA的识别以及与较短靶标的潜在结合。在靶位点将GCG三联体序列改变为ATA，结果清楚地表明Sp1ZF15根据靶序列改变其DNA结合模式。特别有趣的是，Sp1ZF15会根据靶DNA的长度和序列控制参与DNA结合的指状结构域数量。这些结果表明，基于这些多锌指蛋白的人工转录因子在调控大量细胞过程方面具有巨大潜力。
