Direct interaction of p53 with the Y-box binding protein, YB-1: a mechanism for regulation of human gene expression
Direct interaction of p53 with the Y-box binding protein, YB-1: a mechanism for regulation of human gene expression
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DOI：
10.1038/sj.onc.1204029
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发表时间：
2000-12-14
期刊：
影响因子：
8
通讯作者：
Kohno, K
中科院分区：
文献类型：
--
作者：
Okamoto, T;Izumi, H;Kohno, K
Y-box结合蛋白YB-1是一个在转录和翻译水平上调节基因表达的多功能蛋白家族。肿瘤抑制基因p53通过转录调控基因表达而显示出生长抑制特性，我们现在使用体外下拉分析证明YB-1直接与p53相互作用。使用免疫化学共沉淀方法，我们还发现这两种蛋白在体内结合。缺失分析表明，YB-1的三个独立的结构域，一个在N-末端，两个在C-末端，与p53相互作用。凝胶迁移率变动分析表明，YB-1与p53的相互作用刺激了p53与其共有序列的序列特异性DNA结合，相反，这种相互作用抑制了YB-1的结合。在瞬时转染试验中，可以显示YB-1的反义表达抑制p53对该启动子的诱导。这些发现描绘了通过p53-YB-1相互作用的基因表达的直接机制。
The Y-box binding protein, YB-1, belongs to a family of multifunctional proteins which regulate gene expression on both transcriptional and translational levels. The tumor suppressor gene p53 displays growth suppressive properties by regulating gene expression through transcriptional regulation, We now demonstrate that YB-1 directly interacts with p53 using an in vitro pull-down assay. Using immunochemical co-precipitation methods, we also found that the two proteins are bound in vivo. Deletion analysis showed that three independent domains of YB-1, one at the N-terminal and two at the C-terminal, interact with p53. Conversely, a 14 amino acid sequence at the C-terminal of p53 was required for its interaction with YB-1, Gel mobility shift assays showed that the interaction of YB-1 with p53 stimulated the sequence-specific DNA binding of p53 to its consensus sequence, By contrast, this interaction inhibited the binding of YB-1, Using a p53-responsive p21 promoter linked to a reporter gene, it can be shown that antisense expression of YB-1 inhibits the induction of this promoter by p53 in transient transfection assays, These findings delineate a straightforward mechanism for gene expression through p53-YB-1 interaction.