RNA-p53 interactions in vitro.
RNA-p53 interactions in vitro.
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DOI：
10.1021/bi061480v
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发表时间：
2007-02
期刊：
影响因子：
2.9
通讯作者：
K. Riley;M. Ramirez-Alvarado;L. J. Maher
中科院分区：
文献类型：
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作者：
K. Riley;M. Ramirez-Alvarado;L. J. Maher
肿瘤抑制蛋白p53在超过一半的人类癌症中发生突变。尽管经过25年的研究，这种蛋白质的复杂调控仍然不清楚。在酵母三杂交系统（Y3 H）中偶然检测到p53与RNA结合后，我们正在探索这种相互作用的特异性和功能。电泳迁移率变动分析表明，全长p53与Y3 H中强烈区分的RNA同等结合。RNA结合阻断p53与序列特异性DNA的结合。p53的C-末端是体外强RNA相互作用所必需和充分的。小鼠和人C-末端p53肽对RNA具有不同的亲和力，乙酰化的人p53 C-末端肽不结合RNA。p53肽的圆二色性光谱显示RNA结合不会诱导p53 C-末端肽的结构变化，并且C-末端肽不会可检测地影响RNA的结构。这些结果表明，p53结合RNA的序列特异性很小，RNA结合有可能调节DNA结合，和RNA-p53相互作用可以通过乙酰化的p53 C-末端调节。
The tumor suppressor protein p53 is mutated in over half of human cancers. Despite 25 years of study, the complex regulation of this protein remains unclear. After serendipitously detecting RNA binding by p53 in the yeast three-hybrid system (Y3H), we are exploring the specificity and function of this interaction. Electrophoretic mobility shift assays show that full-length p53 binds equally to RNAs that are strongly distinguished in the Y3H. RNA binding blocks sequence-specific DNA binding by p53. The C-terminus of p53 is necessary and sufficient for strong RNA interaction in vitro. Mouse and human C-terminal p53 peptides have different affinities for RNA, and an acetylated human p53 C-terminal peptide does not bind RNA. Circular dichroism spectroscopy of p53 peptides shows that RNA binding does not induce a structural change in the p53 C-terminal peptide, and C-terminal peptides do not detectably affect the structure of RNA. These results demonstrate that p53 binds RNA with little sequence specificity, RNA binding has the potential to regulate DNA binding, and RNA-p53 interactions can be regulated by acetylation of the p53 C-terminus.