Functional characterization of non structural proteins of bovine adenovirus-3

成熟的腺病毒由双链病毒DNA组成，周围是由不同的蛋白质组成的外壳，称为结构蛋白。这种腺病毒的产生还需要非结构蛋白，它不是成熟病毒粒子的一部分，但却是产生新病毒粒子所必需的。牛腺病毒-3感染后期至少有4种非结构蛋白（22K、33K、52K和100K）编码。其中，22K和52K似乎参与了牛腺病毒-3的复制和病毒组装。我们认为22K和52K蛋白参与病毒复制组装的不同步骤可能需要这些蛋白与其他病毒细胞蛋白相互作用。总体目标是鉴定和表征22K和52K与其他病毒细胞蛋白的蛋白-蛋白相互作用，并确定它们在后代牛腺病毒-3生产的不同步骤中的作用。目标将通过a)确定22K和52K与其他病毒蛋白的相互作用，b)确定22K和52K与细胞蛋白的相互作用，c)鉴定和绘制22K和52K与其他病毒细胞蛋白相互作用的结构域来实现。d)通过构建和评估表达突变体（在相互作用结构域缺失）22K或52K蛋白的重组牛腺病毒-3，确定这些蛋白-蛋白相互作用在子代病毒粒子产生中的生物学功能。这项研究将有助于了解22K和52K蛋白在牛腺病毒子代产生中的功能，这将有助于提高牛腺病毒-3以及其他腺病毒的基础生物学知识。此外，拟议研究的研究结果将有助于开发和评估用于动物和人类接种的改良的基于牛腺病毒-3的疫苗递送载体。因此，研究成果将使从事腺病毒教学和工作的学者以及动物疫苗行业受益。

Mature adenovirus consist of double stranded viral DNA surrounded by a shell made up of different proteins called structural proteins. Production of such adenoviruses also require non structural proteins, which are not part of mature virion but are required for the production of new virion. At least four non structural proteins (22K, 33K, 52K and 100K) are encoded at late times post bovine adenovirus-3 infection. Of these, 22K and 52K appear to be involved in bovine adenovirus-3 replication and virus assembly. We believe that involvement of 22K and 52K proteins in different steps of viral replication\assembly may require interaction of these proteins with other viral\cellular proteins.The overall goal is to identify and characterize protein-protein interactions of 22K and 52K with other viral\cellular proteins, and determine their role in different steps of progeny bovine adenovirus-3 production. The goal will be achieved by a) determining interaction of 22K and 52K with other viral proteins, b) determining interaction of 22K and 52K with cellular proteins, c) identifying and mapping the domains of 22K and 52K interacting with other viral \ cellular proteins, d) determining the biological function of these protein-protein interactions in the production of progeny virions by constructing and evaluating recombinant bovine adenovirus-3 expressing mutant (deleted in interacting domain) 22K or 52K proteins. This study will help in understanding the functions of 22K and 52K proteins in the production of progeny bovine adenovirus, which will help in contributing to the advancement of knowledge on fundamental biology of not only bovine adenovirus -3 but adenoviruses in general. Moreover, the research findings of the proposed study will help in developing and evaluating improved bovine adenovirus-3 based vaccine delivery vehicle(s) for vaccination of animals and humans. Thus, the research outcomes will benefit both academicians teaching and working with adenoviruses, and animal vaccine industry.