Foord O S, Bhattacharya P, Reich Z, Rotter V
Department of Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Nucleic Acids Res. 1991 Oct 11;19(19):5191-8. doi: 10.1093/nar/19.19.5191.
In the present study we evaluated the DNA binding activity of wild type and mutant p53 proteins that were isolated from bacterial expression vectors. A comparison of the binding activities of the various purified p53 proteins, assessed by their ability to bind DNA cellulose columns, indicated that wild type p53 has a higher affinity to DNA than have mutant p53 forms. Furthermore, only wild type p53 was able to bind genomic DNA upon electrophoretic protein blotting. As specific deletion of the C-terminal region of wild type p53 totally abolished binding to genomic DNA, it was concluded that the 47 C-terminal amino acids contain the DNA binding region. The fact that the N-terminus contains a transcription activation region whereas the C-terminus contains a DNA binding domain places p53 in the family of typical transcription factors. Our experiments show that the topographical positioning of these domains plays an important role in the activity of wild type p53.
在本研究中，我们评估了从细菌表达载体中分离出的野生型和突变型p53蛋白的DNA结合活性。通过各种纯化的p53蛋白结合DNA纤维素柱的能力来评估其结合活性，结果表明，野生型p53对DNA的亲和力高于突变型p53。此外，只有野生型p53能够在蛋白质印迹电泳时结合基因组DNA。由于野生型p53 C末端区域的特异性缺失完全消除了与基因组DNA的结合，因此得出结论，C末端的47个氨基酸包含DNA结合区域。N末端包含转录激活区域而C末端包含DNA结合结构域这一事实，使p53属于典型转录因子家族。我们的实验表明，这些结构域的拓扑定位在野生型p53的活性中起重要作用。
