PDF(Pubmed)
Abstract：
Plant viruses depend on host cellular factors for their replication and movement. There are cellular proteins that change their localization and/or expression and have a proviral role or antiviral activity and interact with or target viral proteins. Identification of those proteins and their roles during infection is crucial for understanding plant-virus interactions and to design antiviral resistance in crops. Important host proteins have been identified using approaches such as tag-dependent immunoprecipitation or yeast two hybridization that require cloning individual proteins or the entire virus. However, the number of possible interactions between host and viral proteins is immense. Therefore, an alternative method is needed for proteome-wide identification of host proteins involved in host-virus interactions. Here, we present cell fractionation coupled with mass spectrometry as an option to identify protein-protein interactions between viruses and their hosts. This approach involves separating subcellular organelles using differential and/or gradient centrifugation from virus-free and virus-infected cells (1) followed by comparative analysis of the proteomic profiles obtained for each subcellular organelle via mass spectrometry (2). After biological validation, prospect host proteins with proviral or antiviral roles can be subject to fundamental studies in the context of basic biology to shed light on both virus replication and cellular processes. They can also be targeted via gene editing to develop virus-resistant crops.
摘要：
植物病毒的复制和移动依赖于宿主细胞因子。存在改变其定位和/或表达并且具有前病毒作用或抗病毒活性并且与病毒蛋白相互作用或靶向病毒蛋白的细胞蛋白。鉴定这些蛋白质及其在感染过程中的作用对于理解植物-病毒相互作用和设计作物的抗病毒抗性至关重要。重要的宿主蛋白已使用方法如标签依赖性免疫沉淀或需要克隆单个蛋白或整个病毒的酵母2杂交来鉴定。然而,宿主和病毒蛋白之间可能的相互作用数量是巨大的。因此,对于参与宿主-病毒相互作用的宿主蛋白的全蛋白质组鉴定,需要另一种方法.这里,我们将细胞分级分离与质谱联用作为鉴定病毒及其宿主之间蛋白质-蛋白质相互作用的一种选择.该方法涉及使用差异和/或梯度离心从无病毒和病毒感染的细胞中分离亚细胞细胞器(1),然后通过质谱对每个亚细胞细胞器获得的蛋白质组图谱进行比较分析(2)。经过生物验证,具有前病毒或抗病毒作用的前景宿主蛋白可以在基础生物学的背景下进行基础研究，以阐明病毒复制和细胞过程。它们也可以通过基因编辑来开发抗病毒作物。
