Functional characterization of non structural proteins of bovine adenovirus-3

成熟的腺病毒由双链病毒DNA组成，由称为结构蛋白的不同蛋白质组成的外壳包围。这种腺病毒的产生还需要非结构蛋白，其不是成熟病毒体的一部分，但对于新病毒体的产生是必需的。在牛腺病毒-3感染后的晚期，至少编码四种非结构蛋白（22 K、33 K、52 K和100 K）。其中，22 K和52 K似乎参与牛腺病毒-3复制和病毒组装。我们认为22 K和52 K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\细胞蛋白的相互作用，总体目标是鉴定和表征22 K和52 K与其他病毒\细胞蛋白的蛋白质-蛋白质相互作用，并确定它们在子代牛腺病毒3生产的不同步骤中的作用。该目标将通过以下方式实现：a）确定22 K和52 K与其他病毒蛋白的相互作用，B）确定22 K和52 K与细胞蛋白的相互作用，c）鉴定和定位22 K和52 K与其他病毒/细胞蛋白相互作用的结构域，d）确定这些蛋白质的生物学功能-通过构建和评价表达突变体（相互作用结构域缺失）22 K或52 K蛋白的重组牛腺病毒-3，在子代病毒体产生中的蛋白相互作用。本研究将有助于了解22 K和52 K蛋白在子代牛腺病毒产生中的功能，这将有助于促进对牛腺病毒-3乃至一般腺病毒的基础生物学知识的进步。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进的基于牛腺病毒-3的疫苗递送载体。因此，研究成果将有利于教学和工作与腺病毒的学者，和动物疫苗工业。

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.