Functional characterization of structural protein V of bovine adenovirus-3

成熟牛腺病毒-3（BAdV-3）病毒体的产生需要双链病毒DNA和病毒蛋白（非结构和结构）。非结构性病毒蛋白在感染后的早期\晚期表达，但不形成成熟的感染性子代视野的一部分。大多数结构蛋白在感染后的晚期产生，并且是成熟的感染性后代视觉的一部分。其中，蛋白V是一种结构蛋白，其似乎参与核糖体生物发生和维持病毒衣壳的完整性以产生子代BAdV-3。我们认为，BAdV-3蛋白V参与成熟感染性子代病毒体的产生可能需要蛋白质-蛋白质相互作用。事实上，我们最初的实验表明，BAdV-3蛋白V可能与病毒（IVa 2）和细胞（核仁和DDX 21）蛋白相互作用。总体目标是确认和表征这些蛋白质-蛋白质相互作用，并阐明这些相互作用在蛋白质V的不同功能中可能发挥的作用。该目标将通过以下方式实现：a）确认和表征蛋白质V与病毒蛋白质IVa 2的相互作用，B）确认和表征BAdV-3蛋白质V与细胞蛋白质核仁素和DDX 21的相互作用，c）定位i）与IVa 2、核仁和DDX 21相互作用的蛋白V的结构域，ii）与蛋白V相互作用的IVa 2、核仁和DDX 21的结构域，d）通过构建和评估表达突变体的重组牛腺病毒-3，确定这些相互作用在产生成熟感染性子代视野中的生物学意义，本研究的结果将有助于我们理解和阐明这些相互作用在成熟感染性子代BAdV-3产生中的功能。由于蛋白V是乳腺腺病毒属特异性蛋白，因此所提出的研究结果不仅有助于进一步阐明和提高对BAdV-3生物学的认识，而且还有助于了解乳腺腺病毒属的其他成员。此外，研究结果可能为开发和评估用于动物接种的改进的基于BAdV-3的疫苗递送载体铺平道路。因此，学术界（教学\研究）和动物保健行业（包括生产者团体）都将受益。

Production of mature bovine adenovirus-3 (BAdV-3) virions require double stranded viral DNA and viral proteins (non-structural and structural). The non-structural viral proteins are expressed during early\late times post infection but do not form the part of mature infectious progeny visions. Most of the structural proteins are produced at late times post infection and are part of the mature infectious progeny visions. Of these, protein V is a structural proteins which appears to be involved in ribosome biogenesis and in maintaining the integrity of viral capsid for the production of progeny BAdV-3. We believe that involvement of BAdV-3 protein V in the production of mature infectious progeny virions may require protein-protein interactions. In fact, our initial experiments suggest that BAdV-3 protein V may interact with viral (IVa2) and cellular (nucleoli and DDX21) proteins. The overall goal is to confirm and characterize these protein-protein interactions, and elucidate the role these interactions may play in different functions of protein V.The goal will be achieved by a) confirming and characterizing the interaction of protein V with viral protein IVa2, b) confirming and characterizing the interaction of BAdV-3 protein V with cellular proteins nucleolin and DDX21, c) mapping the domains of i) protein V interacting with IVa2, nucleoli and DDX21, ii) IVa2, nucleoli and DDX21 interacting with protein V, d) determining the biological significance of of these interactions in the production of mature infectious progeny visions by constructing and evaluating recombinant bovine adenovirus-3 expressing mutant (interacting domain deleted\substituted) protein V. The results of this study will help us in understanding and elucidating the functions of these interactions in the production of mature infectious progeny BAdV-3. Since protein V is Mastadenovirus genus specific protein, the findings of the proposed research will not only help in further elucidation and advancing the knowledge on BAdV-3 biology, but also about other members of Mastadenovirus genus. In addition, the research findings may pave way for developing and evaluating improved BAdV-3 based vaccine delivery vehicles for animal vaccination. Thus, both academicians (teaching\research) and animal health industry including producer groups will be benefited.