Biochemical specialization within Arabidopsis RNA silencing pathways
Biochemical specialization within Arabidopsis RNA silencing pathways
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DOI：
10.1016/j.molcel.2005.06.014
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发表时间：
2005-08-05
期刊：
影响因子：
16
通讯作者：
Hannon, GJ
中科院分区：
文献类型：
--
作者：
Qi, Y;Denli, AM;Hannon, GJ
在植物中，RNA沉默机制响应于许多输入，包括病毒感染、microRNA和可能以反式和顺式两者起作用的内源性siRNA。此外，已经在植物中证明了沉默结果的全谱，范围从mRNA降解到蛋白质合成水平的抑制到染色质重塑。拟南芥的遗传研究表明，个体的反应途径在功能上是划分的。然而，到目前为止，还没有生物化学系统可用于研究沉默途径中特定蛋白质的作用或选定突变对这些组分的生物化学活性的影响。在这里，我们描述了拟南芥提取物的产生，在体外再现RNA沉默反应的许多方面。我们发现Dicer和Argonaute家族的特定成员具有不同的生化活性，这为深入了解它们在拟南芥RNA沉默途径中的作用提供了依据。
In plants, the RNA silencing machinery responds to numerous inputs, including viral infection, microRNAs, and endogenous siRNAs that may act both in trans and in cis. Additionally, the full spectrum of silencing outcomes has been demonstrated in plants, ranging from mRNA degradation to repression at the level of protein synthesis to chromatin remodeling. Genetic studies in Arabidopsis have indicated that individual response pathways are functionally compartmentalized. However, to date, no biochemical systems have been available to investigate the roles of specific proteins within silencing pathways or the effects of selected mutations on the biochemical activity of those components. Here, we describe the generation of Arabidopsis extracts that reproduce many aspects of RNA silencing reactions in vitro. We find that specific members of the Dicer and Argonaute families have distinct biochemical activities, which provides insight into their roles within RNA silencing pathways in Arabidopsis.