A TRANSCRIPTIONALLY ACTIVE DNA-BINDING SITE FOR HUMAN P53 PROTEIN COMPLEXES
A TRANSCRIPTIONALLY ACTIVE DNA-BINDING SITE FOR HUMAN P53 PROTEIN COMPLEXES
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DOI：
10.1128/mcb.12.6.2866
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发表时间：
1992-06-01
影响因子：
5.3
通讯作者：
SHAY, JW
中科院分区：
文献类型：
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作者：
FUNK, WD;PAK, DT;SHAY, JW
最近的研究表明，肿瘤抑制蛋白p53存在转录激活域，而其他研究则描述了p53的特异性dna结合位点，这意味着该蛋白可能作为转录调节因子。我们使用了重复选择程序（cast：循环扩增和目标选择）来确定p53的新的特异性结合位点，使用正常人类成纤维细胞的核提取物作为p53蛋白的来源。首选的共识是回文GGACATGC CCGGGCATGTCC。在体外翻译的p53只有在与核提取物混合时才能与该序列结合，这表明p53可能在翻译后修饰后与DNA结合，或者与其他蛋白质伙伴形成复合物。当放置在报告结构的上游时，该序列在瞬时转染试验中促进p53依赖性转录。
Recent studies have demonstrated transcriptional activation domains within the tumor suppressor protein p53, while others have described specific DNA-binding sites for p53, implying that the protein may act as a transcriptional regulatory factor. We have used a reiterative selection procedure (CASTing: cyclic amplification and selection of targets) to identify new specific binding sites for p53, using nuclear extracts from normal human fibroblasts as the source of p53 protein. The preferred consensus is the palindrome GGACATGC CCGGGCATGTCC. In vitro-translated p53 binds to this sequence only when mixed with nuclear extracts, suggesting that p53 may bind DNA after posttranslational modification or as a complex with other protein partners. When placed upstream of a reporter construct, this sequence promotes p53-dependent transcription in transient transfection assays.