Foord O, Navot N, Rotter V
Department of Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Mol Cell Biol. 1993 Mar;13(3):1378-84. doi: 10.1128/mcb.13.3.1378-1384.1993.
Wild-type p53 was shown to function as a transcription factor. The N-terminal region of the protein contains the transcription activation domain, while the C terminus is responsible for DNA binding. Localization of the DNA-binding domain of the p53 protein to the highly conserved carboxy-terminal region suggests that the interaction of p53 with DNA is important for its function. We have developed a strategy for studying the DNA sequence specificity of p53-DNA binding that is based on random sequence selection. We report here on the isolation of murine genomic DNA clones that are specifically bound by the wild-type p53 protein but are not bound by mutant p53 protein forms. The isolated p53 target gene contains the unique DNA-binding sequence GACACTGGTCACACTTGGCTGCTTAGGAAT. This fragment exhibits promoter activity as measured by its capacity to activate transcription of the chloramphenicol acetyltransferase reporter gene. Our results suggest that p53 directly binds DNA and functions as a typical transcription factor.
野生型p53被证明可作为一种转录因子发挥作用。该蛋白质的N端区域包含转录激活结构域，而C端负责DNA结合。p53蛋白的DNA结合结构域定位于高度保守的羧基末端区域，这表明p53与DNA的相互作用对其功能很重要。我们开发了一种基于随机序列选择来研究p53-DNA结合的DNA序列特异性的策略。我们在此报告分离出的小鼠基因组DNA克隆，它们能被野生型p53蛋白特异性结合，但不被突变型p53蛋白形式结合。分离出的p53靶基因包含独特的DNA结合序列GACACTGGTCACACTTGGCTGCTTAGGAAT。该片段表现出启动子活性，通过其激活氯霉素乙酰转移酶报告基因转录的能力来衡量。我们的结果表明p53直接结合DNA并作为典型的转录因子发挥作用。
