A genome-wide analysis of the spatio-temporal distribution of herpesviral proteins in the course of infection

感染过程中疱疹病毒蛋白时空分布的全基因组分析

• 批准号: 52364154
• 负责人: Privatdozentin Dr. Susanne Bailer, Ph.D.
• 金额: --
• 依托单位: Max-von-Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2012-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/52364154?language=en
• 关键词: genome; wide; analysis; spatio; temporal

Crucial aspects of the herpesviral life cycle including DNA replication, gene transcription and RNA export, capsid morphogenesis, and egress occur in the host nucleus and require that numerous herpesviral proteins are transported between the host cytosol and the nucleus. The coordinated spatio-temporal distribution of viral proteins between nucleus and cytoplasm is thus vital for viral replication. The docking of incoming herpesviral capsids to the nuclear pore complex (NPC) and the release of viral DNA into the nuclear interior represents a variant of nuclear import and another critical step of viral infection. In general, transport of proteins and some RNAs between cytoplasm and nucleus is mediated by a direct or indirect binding to transport factors of the importin ( and (-families and occurs along a gradient of the small GTPase Ran. So far the mechanism of nucleo-cytoplasmic transport is unknown for a large number of viral proteins and those viral proteins mediating capsid docking have not been identified. In this collaborative project between two groups with high expertise in nucleocytoplasmic transport (S. Bailer) and high-throughput-based screening technologies in infectious diseases (J. Haas), we will apply a novel approach to comprehensively analyse the interactions between herpesviral proteins and all known transport factors of the eukaryotic importin ( and (-families by performing large-scale yeast-two-hybrid (Y2H) screens and in vitro biochemical assays in parallel. The novel viral cargoes of these transport factors will be functionally verified and the subcellular localization of viral proteins and their nuclear transport signals determined systematically. The data received will then be combined by bioinformatical tools to create a dynamic model of the spatio-temporal distribution of viral proteins and capsids during herpesviral infection. This analysis will not only considerably increase our understanding of herpesviral biology and reveal potential novel targets for viral treatment, but also improve the in silico prediction of novel consensus sequences for cellular nuclear transport cargoes.

疱疹病毒生命周期的关键方面，包括DNA复制，基因转录和RNA输出，衣壳形态发生和出口发生在宿主细胞核中，并需要大量疱疹病毒蛋白质在宿主细胞质和细胞核之间转运。因此，病毒蛋白质在细胞核和细胞质之间的协调时空分布对于病毒复制至关重要。进入的疱疹病毒衣壳与核孔复合物（NPC）的对接和病毒DNA释放到核内部代表了核输入的变体和病毒感染的另一个关键步骤。一般来说，蛋白质和一些RNA在细胞质和细胞核之间的转运是通过直接或间接结合到输入蛋白（和）家族的转运因子介导的，并且沿着小GT受体Ran的梯度发生。迄今为止，许多病毒蛋白质的核质转运机制尚不清楚，介导衣壳对接的病毒蛋白质尚未被鉴定。在这个合作项目之间的两个小组具有很高的专业知识，在核质转运（S。Bailer）和感染性疾病高通量筛选技术（J.哈斯）的基础上，我们将采用一种新的方法，通过进行大规模酵母双杂交（Y2H）筛选和体外生化分析，全面分析疱疹病毒蛋白与真核细胞输入素（和）家族所有已知转运因子之间的相互作用。这些转运因子的新型病毒货物将在功能上得到验证，并系统地确定病毒蛋白及其核转运信号的亚细胞定位。然后将通过生物信息学工具将收到的数据结合起来，以创建疱疹病毒感染期间病毒蛋白质和衣壳时空分布的动态模型。这种分析不仅将大大增加我们对疱疹病毒生物学的理解，揭示潜在的病毒治疗新靶点，而且还将改善细胞核转运货物的新型共有序列的计算机预测。

项目成果

Comprehensive Analysis of Varicella-Zoster Virus Proteins Using a New Monoclonal Antibody Collection

使用新的单克隆抗体系列对水痘带状疱疹病毒蛋白进行综合分析

• DOI: 10.1128/jvi.00407-13
• 发表时间: 2013
• 期刊: Journal of Virology
• 影响因子: 5.4
• 作者: Bailer;S. M. ;Pothineni;Ouwendijk;W. J. D;Endesfelder;Bajdaric;Baiker;Verjans;G. M. G. M;Jonjic
• 通讯作者: Jonjic