Functional characterization of non structural proteins of bovine adenovirus 3

腺病毒由裸露的二十面体衣壳、末端重复序列倒置的双链DNA基因组和附着在基因组5‘端的多肽组成。腺病毒至少编码三种非结构蛋白，它们被认为在不同启动子的反式激活、晚期mRNAs的成熟和翻译、DNA包膜和病毒衣壳组装中发挥作用。牛腺病毒(BAdV)-3的非结构蛋白为52K、100K和33K。其中100K和33K是磷酸核蛋白，它们似乎是BAdV-3复制所必需的，并可能参与DNA包装、病毒衣壳组装和有效的DNA/衣壳相互作用。我们认为，100K和33K参与BAdV-3病毒组装的不同方面可能需要这些蛋白与其他病毒/细胞蛋白相互作用。这项研究的假设是非结构蛋白包含多种功能，这对病毒的复制和组装非常重要。此外，这些蛋白质执行多功能的能力可能需要与其他病毒/细胞蛋白质相互作用(S)。因此，总的目标是确定、表征、定位和确定33K和100K在后代病毒粒子生产中执行不同功能的机制。这项资助的具体目标是详细研究33K和100K与其他病毒/细胞蛋白的相互作用，这将有助于确定这些非结构蛋白执行不同功能的机制。具体目标将通过a)调查33K和100K与其他病毒蛋白的相互作用，b)调查100K和33K与细胞蛋白的相互作用，c)绘制与其他病毒/细胞蛋白相互作用所需的33K和100K结构域，以及d)确定已确定的蛋白质-蛋白质相互作用对BAdV-3复制的重要性。

Adenovirus consists of a naked icosahedral capsid, double stranded DNA genome with inverted terminal repeats and a polypeptide attached to the 5' ends of the genome. At least three non structural proteins are encoded by adenoviruses, which are believed to function in transactivation of different promoters, maturation and translation of late mRNAs, DNA encapsidation and viral capsid assembly. In bovine adenovirus (BAdV) -3, the non structural proteins are 52K, 100K and 33K. Of these, 100K and 33K are phosphonuclear proteins, which appear to be essential for BAdV-3 replication and, may be involved in DNA packaging, viral capsid assembly and efficient DNA/capsid interaction. We believe that involvement of 100K and 33K in different aspects of BAdV-3 virus assembly may require interaction of these proteins with other viral/cellular proteins. The hypothesis for this study is that non structural proteins contain multi-functions, which are important for viral replication and assembly. Moreover, ability of these proteins to perform multi-functions may require interactions with other viral / cellular protein(s). So the overall goal is to identify, characterize, map and determine the mechanism of performing different functions of 33K and 100K in the production of progeny virions. The specific goal of this grant is to investigate in detail the interaction of 33K and 100K with other viral/cellular proteins, which will help in determining the mechanism of performing different functions by these non structural proteins. The specific goal will be achieved by a) investigating the interaction of 33K and 100K with other viral proteins, b) investigating the interaction of 100K and 33K with cellular proteins, c) mapping the domains of 33K and 100K required for the interaction with other viral / cellular proteins and d) determining the importance of identified protein-protein interaction for BAdV-3 replication.