SEQUENCE-SPECIFIC DNA-BINDING BY A SHORT PEPTIDE DIMER
SEQUENCE-SPECIFIC DNA-BINDING BY A SHORT PEPTIDE DIMER
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DOI：
10.1126/science.2389142
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发表时间：
1990-08-17
期刊：
影响因子：
56.9
通讯作者：
KIM, PS
中科院分区：
文献类型：
--
作者：
TALANIAN, RV;MCKNIGHT, CJ;KIM, PS
最近描述的一类DNA结合蛋白的特征在于“bZIP”基序，其由接触DNA的碱性区域和介导蛋白质二聚化的相邻“亮氨酸拉链”组成。已经开发了酵母转录激活因子GCN 4的碱性区域的肽模型，其中亮氨酸拉链已被二硫键取代。34-残基肽二聚体，而不是还原的单体，以纳摩尔亲和力在4 ° C结合DNA。C. DNA结合是序列特异性的，如通过足迹法中的脱氧核糖核酸酶所判断的。圆二色光谱表明，该肽采用螺旋结构时，结合到DNA。这些结果直接表明，GCN 4碱性区是足够的序列特异性DNA结合，并表明GCN 4亮氨酸拉链的主要功能是简单地介导蛋白质二聚化。我们的方法提供了一种策略，设计短序列特异性的DNA结合肽。
A recently described class of DNA binding proteins is characterized by the "bZIP" motif, which consists of a basic region that contacts DNA and an adjacent "leucine zipper" that mediates protein dimerization. A peptide model for the basic region of the yeast transcriptional activator GCN4 has been developed in which the leucine zipper has been replaced by a disulfide bond. The 34-residue peptide dimer, but not the reduced monomer, binds DNA with nanomolar affinity at 4.degree. C. DNA binding is sequence-specific as judged by deoxyribonuclease in footprinting. Circular dichroism spectroscopy suggests that the peptide adopts a helical structure when bound to DNA. These results demonstrate directly that the GCN4 basic region is sufficient for sequence-specific DNA binding and suggest that a major function of the GCN4 leucine zipper is simply to mediate protein dimerization. Our approach provides a strategy for the design of short sequence-specific DNA binding peptides.