An RNA aptamer that interferes with the DNA binding of the HSF transcription activator.
An RNA aptamer that interferes with the DNA binding of the HSF transcription activator.
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DOI：
10.1093/nar/gkl470
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发表时间：
2006
影响因子：
14.9
通讯作者：
Lis JT
中科院分区：
文献类型：
--
作者：
Zhao X;Shi H;Sevilimedu A;Liachko N;Nelson HC;Lis JT
热休克因子（HSF）是一种保守的高效转录激活因子。它参与各种重要的生物过程，包括应激反应和正常发育中的特定步骤。干扰HSF功能的试剂对于基础研究和实际应用都是有用的。我们选择了一个RNA适体，结合HSF具有高特异性。缺失分析确定该适体的最小结合基序为两个茎和一个茎-环，其通过三向连接连接。这种RNA适体干扰HSF与其DNA元件的正常相互作用，这是HSF功能的关键调节步骤。DNA结合结构域加上HSF（DL）上的侧翼接头区对于RNA结合是必需的。此外，这种适体在体外抑制在全细胞提取物的复杂环境中的HSF诱导的转录。与先前表征的NF-κB适体相反，HSF适体不简单地模拟DNA结合，而是以不同于DNA结合HSF的方式结合HSF。
Heat shock factor (HSF) is a conserved and highly potent transcription activator. It is involved in a wide variety of important biological processes including the stress response and specific steps in normal development. Reagents that interfere with HSF function would be useful for both basic studies and practical applications. We selected an RNA aptamer that binds to HSF with high specificity. Deletion analysis defined the minimal binding motif of this aptamer to be two stems and one stem–loop joined by a three-way junction. This RNA aptamer interferes with normal interaction of HSF with its DNA element, which is a key regulatory step for HSF function. The DNA-binding domain plus a flanking linker region on the HSF (DL) is essential for the RNA binding. Additionally, this aptamer inhibits HSF-induced transcription in vitro in the complex milieu of a whole cell extract. In contrast to the previously characterized NF-κB aptamer, the HSF aptamer does not simply mimic DNA binding, but rather binds to HSF in a manner distinct from DNA binding to HSF.