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Investigation of the effects of P1 on HC-pro-mediated gene silencing suppression through genetics and omics approaches

机译:P1对通过遗传学和常规方法的HC-Pro介导的基因沉默抑制的研究

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BACKGROUND:Posttranscriptional gene silencing (PTGS) is one of the most important mechanisms for plants during viral infection. However, viruses have also developed viral suppressors to negatively control PTGS by inhibiting microRNA (miRNA) and short-interfering RNA (siRNA) regulation in plants. The first identified viral suppressor, P1/HC-Pro, is a fusion protein that was translated from potyviral RNA. Upon infecting plants, the P1 protein itself is released from HC-Pro by the self-cleaving activity of P1. P1 has an unknown function in enhancing HC-Pro-mediated PTGS suppression. We performed proteomics to identify P1-interacting proteins. We also performed transcriptomics that were generated from Col-0 and various P1/HC-Pro-related transgenic plants to identify novel genes. The results showed several novel genes were identified through the comparative network analysis that might be involved in P1/HC-Pro-mediated PTGS suppression.RESULTS:First, we demonstrated that P1 enhances HC-Pro function and that the mechanism might work through P1 binding to VERNALIZATION INDEPENDENCE 3/SUPERKILLER 8 (VIP3/SKI8), a subunit of the exosome, to interfere with the 5'-fragment of the PTGS-cleaved RNA degradation product. Second, the AGO1 was specifically posttranslationally degraded in transgenic Arabidopsis expressing P1/HC-Pro of turnip mosaic virus (TuMV) (P1/HC Tu plant). Third, the comparative network highlighted potentially critical genes in PTGS, including miRNA targets, calcium signaling, hormone (JA, ET, and ABA) signaling, and defense response.CONCLUSION:Through these genetic and omics approaches, we revealed an overall perspective to identify many critical genes involved in PTGS. These new findings significantly impact in our understanding of P1/HC-Pro-mediated PTGS suppression.
机译:背景:后术后基因沉默(PTG)是病毒感染期间植物中最重要的机制之一。然而,病毒还通过抑制MicroRNA(miRNA)和植物中的短干扰RNA(siRNA)调节来开发病毒抑制剂以消除PTG。第一个鉴定的病毒抑制器P1 / HC-Pro是融合蛋白,其由potyViral RNA翻译。在感染植物后,通过P1的自切割活性从HC-Pro释放P1蛋白本身。 P1在增强HC-Pro介导的PTGS抑制方面具有未知功能。我们进行了蛋白质组学以鉴定P1相互作用的蛋白质。我们还进行了从COL-0和各种P1 / HC-PRO相关的转基因植物产生的转录组科以鉴定新基因。结果表明,通过比较网络分析鉴定了几种新基因,这些基因可能涉及p1 / hc-pro介导的ptgs抑制。结果:首先,我们证明了P1增强了HC-Pro功能,并且该机制可以通过P1绑定工作对于vernalizate独立性3 /超级铝偶8(VIP3 / SKI8),外出组的亚基,以干扰PTGS切割的RNA降解产物的5'-片段。其次,前一个特异性地在表达萝卜叶片病毒(Tumv)的P1 / HC-Pro(P1 / HC Tu植物)中的转基因拟南芥中的转基因拟南芥中降解。第三,比较网络突出显示PTG的潜在关键基因,包括miRNA靶,钙信号,激素(JA,ET和ABA)信号传导,防御反应。结论:通过这些遗传和常规方法,我们揭示了识别的整体视角涉及PTG的许多关键基因。这些新发现对我们对P1 / HC-Pro-Pro介导的PTGS抑制的理解显着影响。

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