Host cell substrates of RNA virus proteases

双环病毒是一种主要侵染节肢动物的RNA病毒，对农业和经济都有负面影响。成员包括与蜜蜂疾病有关的蜜蜂病毒和导致泛美白对虾行业疾病爆发的陶拉综合征病毒。到目前为止，对病毒的生命周期和宿主之间的相互作用还没有完全了解，因此影响了我们对这些病毒的理解和有限的潜在抗病毒策略。然而，果蝇(果蝇)被双环病毒蟋蟀麻痹病毒(CRPV)感染在破译双环病毒如何侵占宿主细胞的关键过程中发挥了重要作用。双胞菌病毒的标志性特征之一是，该病毒表达一种名为蛋白酶的蛋白质，该蛋白质在细胞内裂解宿主蛋白以促进病毒感染。目前，病毒蛋白水解酶靶向宿主蛋白的程度尚不清楚。在这项研究中，我们将利用一种名为TILS的新的无偏倚蛋白质组学方法来全面鉴定DicistroVirus CRPV蛋白酶在果蝇细胞中的所有靶点。与其他方法不同，这种新的尾巴方法是敏感的，可以确定目标蛋白上切割位点的位置。新靶点的确定将提供在双胞菌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。这一策略的发展可以被用来从其他病毒中识别蛋白酶的宿主靶标，最终导致对一系列导致病理疾病的病毒感染的更好的治疗方案。

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.