Host cell substrates of RNA virus proteases

双顺反子病毒是RNA病毒，主要感染节肢动物，并对农业和经济重要性产生负面影响。成员包括与蜜蜂疾病有关的蜜蜂病毒和导致对虾产业疾病爆发的Taura综合征病毒。迄今为止，生命周期和病毒宿主相互作用尚未完全了解，因此，影响了我们对这些病毒的理解，并限制了潜在的抗病毒策略。然而，通过模型双顺反子病毒蟋蟀麻痹病毒（CrPV）感染果蝇（果蝇）已经有助于破译双顺反子病毒如何霸占宿主细胞的关键过程。双顺反子病毒的特征之一是病毒表达一种称为蛋白酶的蛋白质，该蛋白酶切割细胞中的宿主蛋白质以促进病毒感染。目前，病毒蛋白酶靶向宿主蛋白的程度尚不清楚。在这项研究中，我们将利用一种新的无偏见的蛋白质组学方法，称为TAILS，全面确定果蝇细胞中的双顺反子病毒CrPV蛋白酶的所有目标。与其他方法不同，这种新的TAILS方法是敏感的，可以识别靶蛋白上切割位点的位置。新靶标的鉴定将提供在双顺反子病毒感染期间受影响的宿主过程的目录。重要的是，所获得的知识将使我们能够确定抗病毒治疗的新靶点。这种策略的发展可以用于识别来自其他病毒的蛋白酶的宿主靶标的库，最终导致更好的治疗方案用于一系列引起病理性疾病的病毒感染。

Dicistroviruses are RNA viruses that mainly infect arthropods and have had a negative impact of agriculture and economic importance. Members include the honeybee viruses that have been associated with honeybee disease and the Taura syndrome virus, which has led to disease outbreaks in the panaeid shrimp industry. To date, the life cycle and virus host interactions are not fully understood and thus, has impacted our understanding of these viruses and limited potential antiviral strategies. However, infection of Drosophila (fruit fly) by the model dicistrovirus cricket paralysis virus (CrPV) has been instrumental in deciphering key processes into how dicistroviruses commandeer the host cell. One of the signature properties of dicistroviruses is that the virus expresses a protein, called a protease, which cleaves host proteins in the cell to facilitate virus infection. Currently, the extent of host proteins targeted by the viral protease is not known. In this study, we will utilize a novel unbiased proteomics approach called TAILS to comprehensively identify all of the targets of the dicistrovirus CrPV protease in Drosophila cells. Unlike other methods, this new TAILS approach is sensitive and can identify the location of the cleavage site on the target protein. The identification of novel targets will provide a catalog of the host processes that are affected during dicistrovirus infection. Importantly, the knowledge gained will allow us to identify new targets for antiviral therapy. The development of this strategy can be exploited to identify the repertoire of host targets of proteases from other viruses, ultimately leading to better treatment protocols for a range of virus infections causing pathological disease.