Bortolamiol Diane, Pazhouhandeh Maghsoud, Marrocco Katia, Genschik Pascal, Ziegler-Graff Véronique
Institut de Biologie Moléculaire des Plantes du CNRS, 12 rue du Général Zimmer, 67084 Strasbourg, France.
Curr Biol. 2007 Sep 18;17(18):1615-21. doi: 10.1016/j.cub.2007.07.061.
Plants employ post-transcriptional gene silencing (PTGS) as an antiviral defense response. In this mechanism, viral-derived small RNAs are incorporated into the RNA-induced silencing complex (RISC) to guide degradation of the corresponding viral RNAs. ARGONAUTE1 (AGO1) is a key component of RISC: it carries the RNA slicer activity. As a counter-defense, viruses have evolved various proteins that suppress PTGS. Recently, we showed that the Polerovirus P0 protein carries an F box motif required to form an SCF-like complex, which is also essential for P0's silencing suppressor function. Here, we investigate the molecular mechanism by which P0 impairs PTGS. First we show that P0's expression does not affect the biogenesis of primary siRNAs in an inverted repeat-PTGS assay, but it does affect their activity. Moreover, P0's expression in transformed Arabidopsis plants leads to various developmental abnormalities reminiscent of mutants affected in miRNA pathways, which is accompanied by enhanced levels of several miRNA-target transcripts, suggesting that P0 acts at the level of RISC. Interestingly, ectopic expression of P0 triggered AGO1 protein decay in planta. Finally, we provide evidence that P0 physically interacts with AGO1. Based on these results, we propose that P0 hijacks the host SCF machinery to modulate gene silencing by destabilizing AGO1.
植物利用转录后基因沉默(PTGS)作为一种抗病毒防御反应。在这种机制中,病毒衍生的小RNA被整合到RNA诱导沉默复合体(RISC)中,以指导相应病毒RNA的降解。AGO1是RISC的关键组成部分:它具有RNA切割活性。作为一种反击手段,病毒进化出了多种抑制PTGS的蛋白质。最近,我们发现马铃薯卷叶病毒P0蛋白带有一个形成类似SCF复合体所需的F盒基序,这对P0的沉默抑制功能也至关重要。在这里,我们研究了P0损害PTGS的分子机制。首先,我们发现在反向重复PTGS试验中,P0的表达不影响初级siRNA的生物合成,但会影响其活性。此外,P0在转化的拟南芥植物中的表达会导致各种发育异常,这让人联想到在miRNA途径中受影响的突变体,同时伴随着几种miRNA靶转录本水平的提高,这表明P0在RISC水平上起作用。有趣的是,P0的异位表达在植物中引发了AGO1蛋白的降解。最后,我们提供证据表明P0与AGO1存在物理相互作用。基于这些结果,我们提出P0通过劫持宿主SCF机制,使AGO1不稳定来调节基因沉默。