Janknecht R, Nordheim A
Institute for Molecular Biology, Hannover Medical School, Germany.
Nucleic Acids Res. 1992 Jul 11;20(13):3317-24. doi: 10.1093/nar/20.13.3317.
The Ets-related Elk-1 protein can bind to purine-rich DNA target sites in a sequence specific fashion and, in addition, can form a ternary complex with the c-fos serum response element (SRE) and the serum response factor (SRF). We demonstrate that Elk-1 can readily interchange between its different interaction partners. The amino terminal ETS-domain of Elk-1 was shown to be necessary and sufficient for direct DNA-binding activity. For ternary complex formation with the SRE and SRF, both the Elk-1 ETS-domain as well as flanking sequences up to amino acid 169 were required. Removal of sequences between the ETS-domain and amino acids 137-169 did not abolish ternary complex formation. This suggests the Elk-1 region spanning amino acids 137-169 to contain a protein-protein interaction domain. Furthermore, we have shown that a single amino acid exchange introduced into the ETS-domain can drastically alter the direct DNA-binding affinity of Elk-1 without severely affecting SRF-assisted binding to the SRE. Thus, Elk-1 requires different propensities of the ETS-domain to exert its different modes of DNA sequence recognition.
Ets相关的Elk-1蛋白能够以序列特异性的方式结合富含嘌呤的DNA靶位点,此外,它还能与c-fos血清反应元件(SRE)和血清反应因子(SRF)形成三元复合物。我们证明Elk-1能够在其不同的相互作用伙伴之间轻易互换。Elk-1的氨基末端ETS结构域被证明对于直接DNA结合活性是必要且充分的。对于与SRE和SRF形成三元复合物,Elk-1的ETS结构域以及直至第169位氨基酸的侧翼序列都是必需的。去除ETS结构域与第137 - 169位氨基酸之间的序列并不会消除三元复合物的形成。这表明Elk-1中跨越第137 - 169位氨基酸的区域包含一个蛋白质 - 蛋白质相互作用结构域。此外,我们已经表明,引入到ETS结构域中的单个氨基酸交换能够极大地改变Elk-1的直接DNA结合亲和力,而不会严重影响SRF辅助的与SRE的结合。因此,Elk-1需要ETS结构域具有不同的倾向来发挥其不同的DNA序列识别模式。