Comprehensive DNA recognition through concerted interactions from adjacent zinc fingers
Comprehensive DNA recognition through concerted interactions from adjacent zinc fingers
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DOI：
10.1021/bi981358z
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发表时间：
1998-09-01
期刊：
影响因子：
2.9
通讯作者：
Choo, Y
中科院分区：
文献类型：
--
作者：
Isalan, M;Klug, A;Choo, Y
锌指是一种小型的DNA结合模块，因其在大量真核转录因子中的存在以及在蛋白质工程中的应用而受到关注。尽管人们预期锌指能够结合多种多样的DNA序列，但先前使用来自Zif268（和Sp1）的锌指结构域模型所进行的研究揭示了DNA结合特异性存在潜在的局限性。例如，通过噬菌体展示选择单个锌指来识别三核苷酸DNA亚位点，得到的锌指仅特异性结合形式为GNN的三联体，即5'端为鸟嘌呤的三联体。在我们最近报道的工作[伊萨兰，M.；周，Y.；克鲁格，A.（1997）《美国国家科学院院刊》94，5617 - 5621]之后，我们现在表明，通过对指定重叠DNA亚位点的相邻锌指中某些氨基酸位置进行协同随机化，可以克服这种局限性。这阐明了锌指阵列识别DNA的一个重要机制，并为设计能结合任何给定核苷酸序列的高特异性锌指蛋白的改进策略指明了方向。
Zinc fingers are small DNA-binding modules noted for their occurrence in a large number of eukaryotic transcription factors, and their use in protein engineering. Although it was expected that zinc fingers can bind to a wide diversity of DNA sequences, previous studies using model zinc finger domains from Zif268 (and Sp1) have revealed a potential Limitation to the DNA-binding specificity. For example, phage display selection of individual zinc fingers to recognize trinucleotide DNA subsites returned fingers that bound specifically only to triplets of the form GNN, i.e., triplets with guanine at the 5' end. Following our recently reported work [Isalan, M., Choo, Y., and Klug, A. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 5617-5621], we now show that this limitation can be overcome by the concerted randomization of certain amino acid positions in adjacent zinc fingers that specify overlapping DNA subsites. This illustrates an important mechanism underlying DNA recognition by arrays of zinc fingers, and points the way to improved strategies for the design of highly specific zinc finger proteins that bind any given nucleotide sequence.