Stimulators of translation identified during a small molecule screening campaign.
Stimulators of translation identified during a small molecule screening campaign.
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在小分子筛选活动中发现的翻译刺激物。
DOI：
10.1016/j.ab.2013.10.026
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发表时间：
2014
影响因子：
2.9
通讯作者：
Pelletier,Jerry
中科院分区：
文献类型：
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作者：
Shin,Unkyung;Williams,DavidE;Kozakov,Dima;Hall,DavidR;Beglov,Dmitri;Vajda,Sandor;Andersen,RaymondJ;Pelletier,Jerry
在筛选真核翻译调节剂的天然和合成产物库时，我们鉴定了两种天然产物，异膜醇二胺和膜醇二胺，它们对翻译具有刺激作用。这些化合物的表征使我们认识到，在高通量分析设置过程中，用于编程翻译提取物的mRNA导致了eIF2α的磷酸化，这是一个由四种激酶之一介导的强有力的负调控事件。我们发现双链rna依赖蛋白激酶（PKR）是外源添加mRNA模板激活的eIF2α激酶。对其作用方式的表征表明，它通过抑制PKR的自磷酸化直接作用于PKR，扰乱PKR - eif2 α磷酸化轴，并可建模为PKR ATP结合位点。我们的研究结果确定了使用翻译摘录的高通量屏幕活动的“假阳性”来源，对这类屏幕提出了警告。
In screening a library of natural and synthetic products for eukaryotic translation modulators, we identified two natural products, isohymenialdisine and hymenialdisine, that exhibit stimulatory effects on translation. The characterization of these compounds led to the insight that mRNA used to program the translation extracts during high-throughput assay setup was leading to phosphorylation of eIF2α, a potent negative regulatory event that is mediated by one of four kinases. We identified double-stranded RNA-dependent protein kinase (PKR) as the eIF2α kinase that was being activated by exogenously added mRNA template. Characterization of the mode of action of isohymenialdisine revealed that it directly acts on PKR by inhibiting autophosphorylation, perturbs the PKR–eIF2α phosphorylation axis, and can be modeled into the PKR ATP binding site. Our results identify a source of “false positives” for high-throughput screen campaigns using translation extracts, raising a cautionary note for this type of screen.