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'末端的多肽组成。腺病毒编码至少三种非结构蛋白，这些蛋白被认为在不同启动子的反激活、后期mrna的成熟和翻译、DNA封装和病毒衣壳组装中起作用。在牛腺病毒(BAdV) -3中，非结构蛋白为52K、100K和33K。其中，100K和33K是磷酸核蛋白，对badv3的复制至关重要，可能参与DNA包装、病毒衣壳组装和DNA/衣壳的有效相互作用。我们认为100K和33K参与badv3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。本研究假设非结构蛋白具有多种功能，对病毒的复制和组装具有重要作用。此外，这些蛋白执行多种功能的能力可能需要与其他病毒/细胞蛋白相互作用。因此，我们的总体目标是鉴定、表征、绘制和确定33K和100K在子代病毒粒子产生中发挥不同功能的机制。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)研究33K和100K与其他病毒蛋白的相互作用，b)研究100K和33K与细胞蛋白的相互作用，c)绘制33K和100K与其他病毒/细胞蛋白相互作用所需的结构域，以及d)确定所鉴定的蛋白-蛋白相互作用对badv3复制的重要性来实现。本研究的意义在于了解badv3的100K和33K蛋白的功能机制，这可能有助于开发用于人类和动物基因传递和疫苗接种的改良载体。

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.The significance of this study lies in understanding the mechanism of functions performed by 100K and 33K proteins of BAdV-3, which may lead to the development of improved vectors for gene delivery and vaccination of humans and animals.