A PROP1-binding factor, AES cloned by yeast two-hybrid assay represses PROP1-induced Pit-1 gene expression.
A PROP1-binding factor, AES cloned by yeast two-hybrid assay represses PROP1-induced Pit-1 gene expression.
复制标题
通过酵母双杂交测定克隆的 PROP1 结合因子 AES 可抑制 PROP1 诱导的 Pit-1 基因表达。
DOI：
10.1016/j.mce.2013.05.022
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
Okimura Y.
中科院分区：
文献类型：
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作者：
Sugiyama Y;Ikeshita N;Shibahara H;Yamamoto D;Kawagishi M;Iguchi G;Iida K;Takahashi Y;Kaji H;Chihara K;Okimura Y.
PROP 1突变导致垂体激素缺乏症（CPHD）。几个突变位于Prop 1的反式激活结构域（transactivation domain，CET），CET与辅因子结合的丧失可能是CPHD的原因。PROP 1辅因子尚未确定。在本研究中，我们的目的是确定PROP 1相互作用的蛋白质从人脑cDNA文库。使用酵母双杂交试验，我们克隆了九个候选蛋白，可能会结合到PROP 1。在这九个候选者中，氨基末端分裂增强剂（AES）是最丰富的，我们分析了AES功能。AES剂量依赖性地降低PROP 1诱导的Pit-1报告基因表达。免疫沉淀试验揭示了AES和PROP 1之间的关系。在哺乳动物的双杂交试验中，AES Q结构域的亮氨酸拉链样基序被鉴定为与p53相互作用的区域。这些结果表明AES是PROP 1的辅阻遏物。
PROP1 mutation causes combined pituitary hormone deficiency (CPHD). Several mutations are located in a transactivation domain (TAD) of Prop1, and the loss of TAD binding to cofactors is likely the cause of CPHD. PROP1 cofactors have not yet been identified. In the present study, we aimed to identify the PROP1-interacting proteins from the human brain cDNA library. Using a yeast two-hybrid assay, we cloned nine candidate proteins that may bind to PROP1. Of those nine candidates, amino-terminal enhancer of split (AES) was the most abundant, and we analyzed the AES function. AES dose-dependently decreased the PROP1-induced Pit-1 reporter gene expression. An immunoprecipitation assay revealed the relationship between AES and PROP1. In a mammalian two-hybrid assay, a leucine zipper-like motif of the AES Q domain was identified as a region that interacted with TAD. These results indicated that AES was a corepressor of PROP1.