A DNA-BINDING DOMAIN IS CONTAINED IN THE C-TERMINUS OF WILD-TYPE P53-PROTEIN
A DNA-BINDING DOMAIN IS CONTAINED IN THE C-TERMINUS OF WILD-TYPE P53-PROTEIN
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DOI：
10.1093/nar/19.19.5191
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发表时间：
1991-10-11
影响因子：
14.9
通讯作者：
ROTTER, V
中科院分区：
文献类型：
--
作者：
FOORD, OS;BHATTACHARYA, P;ROTTER, V
在本研究中，我们评估了从细菌表达载体中分离的野生型和突变型p53蛋白的DNA结合活性。比较各种纯化的p53蛋白的结合活性，通过它们结合DNA纤维素柱的能力进行评估，表明野生型p53比突变型p53对DNA具有更高的亲和力。此外，只有野生型p53能够结合基因组DNA电泳蛋白印迹。由于野生型p53的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，47个C-末端氨基酸含有DNA结合区域。事实上，N-末端含有一个转录激活区，而C-末端含有一个DNA结合结构域，将p53置于典型的转录因子家族中。我们的实验表明，这些结构域的地形定位在野生型p53的活性中起着重要的作用。
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.