Defining the nuclear vs. cytoplasmic proteome during human rhinovirus infections

小核糖核酸病毒是人类的主要病原体，它们在世界范围内的流行是导致 是所有已知病毒中发病率最高的。由于编码容量有限， 微小核糖核酸病毒基因组RNA、宿主蛋白在病毒翻译和RNA中起着关键作用 在感染细胞的细胞质中复制。这些蛋白质中的许多通常是定位的。 未受感染的人类细胞的细胞核。本申请中提出的实验旨在 确定人类鼻病毒感染细胞中所带来的变化的蛋白质图景 通过病毒诱导的宿主细胞核质运输机制的改变。 该提案将探索鼻病毒进化利用的独特机制 在IRES介导的翻译和病毒RNA中起关键作用的宿主核蛋白 在其细胞内复制周期中的合成。通过与正常的核居住者互动 蛋白质在其细胞质复制周期中，鼻病毒扩大宿主细胞的保留范围 它可以为自己的目的劫持功能。使用一种公正的蛋白质组学方法，这 应用程序将解决人类鼻病毒诱导宿主重新分区的假设 在细胞核和细胞质之间的细胞蛋白质，以加强其复制周期中的步骤。 拟议的实验将扩大最近的一项发现，即鼻病毒感染 人肺成纤维细胞导致了一套不同的病毒-宿主相互作用 在鼻病毒或脊髓灰质炎病毒感染HeLa细胞期间观察到。两个实验目标将 以发现人类鼻病毒是否使用一组不同的宿主蛋白和 肺细胞复制过程中的机制，部分是由新的病毒诱导的 驻留的核蛋白重新分布到感染细胞的细胞质：(1)产生一个 人鼻病毒感染过程中核质与细胞质蛋白质组的比较，以及(2) 人鼻病毒候选宿主蛋白的鉴定及其功能意义的确定 复制。总体而言，确定鼻病毒和其他小核糖核酸病毒如何利用 改变的核质运输应该揭示对RNA病毒的新的机制洞察 复制并在病毒-宿主界面确定抗病毒治疗的新靶点。

Picornaviruses are major human pathogens, and their worldwide prevalence is responsible for some of the highest morbidities of all known viruses. Due to the limited coding capacity of picornavirus genomic RNAs, host proteins play critical roles during viral translation and RNA replication in the cytoplasm of infected cells. A number of these proteins are normally localized to the nucleus of uninfected human cells. The experiments proposed in this application aim to define the changing protein landscape in human rhinovirus infected cells that is brought about by viral-induced modifications of the nucleo-cytoplasmic trafficking machinery of the host cell. The proposal will explore the unique mechanisms that rhinovirus has evolved to take advantage of host nuclear proteins which play key roles in IRES-mediated translation and viral RNA synthesis during its intracellular replication cycle. By interacting with normally nuclear-resident proteins during its cytoplasmic replication cycle, rhinovirus expands the repertoire of host cell functions it can hijack for its own purposes. Using an unbiased proteomics approach, this application will address the hypothesis that human rhinovirus induces a re-partitioning of host cell proteins between the nucleus and the cytoplasm to potentiate steps in its replication cycle. The proposed experiments will expand upon a recent discovery that rhinovirus infection of human lung fibroblast cells results in a different set of virus-host interactions compared to what is observed during a rhinovirus or poliovirus infection of HeLa cells. Two experimental aims will be carried out to discover if human rhinovirus uses a distinct set of host proteins and mechanisms during replication in lung cells aided, in part, by the novel viral-induced redistribution of resident nuclear proteins to the cytoplasm of infected cells: (1) Generate a nuclear versus cytoplasmic human cell proteome during a human rhinovirus infection, and (2) Identify candidate host proteins and determine their functional significance for human rhinovirus replication. Overall, determining how rhinoviruses and other picornaviruses take advantage of altered nucleo-cytoplasmic trafficking should reveal new mechanistic insights into RNA virus replication and identify novel targets for antiviral therapeutics at the virus-host interface.