Functional characterization of BHV-1 tegument proteins

BHV-1 被膜蛋白的功能表征

• 批准号: 90887-2010
• 负责人: VanDrunenLittelvandenHurk, Sylvia
• 金额: $ 4.08万
• 依托单位: University of Saskatchewan
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2012
• 资助国家: 加拿大
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=513561
• 关键词: Functional; characterization; BHV; tegument; proteins

Herpesviruses are composed of four concentric compartments, the nucleoprotein core surrounding the DNA, the capsid, the tegument and the envelope. The tegument contains at least 15 proteins, which play an important role in viral infection, because they are the first to interact with the intracellular environment and at a later stage they associate to create new virions. The long-term objective of this program is to perform a functional characterization of the tegument proteins of bovine herpesvirus-1 (BHV-1), one of the major pathogens in bovine respiratory disease. The specific objective of this proposal is to further characterize the functions of VP8, which is the major tegument protein in BHV-1. Additional tegument proteins, in particular those that interact with VP8 will be studied following similar approaches. A mutant virus lacking VP8 was impaired in vitro and non-virulent in vivo, which suggests that in addition to a structural role VP8 has an important early function, possibly modulating innate immune responses. Our hypothesis is that VP8 plays a role both in establishing conditions for efficient viral replication and in promoting virus assembly. We will test this as follows: i) Since VP8 has RNA binding activity, we will examine the early effects of VP8 on viral mRNA transport; ii) Interestingly, VP8 interacts with DNA damage-binding protein-1 and culllin4A, so we will determine whether VP8 hijacks the cullin-4A-DDB1 E3 ubiquitin ligase to control protein degradation in infected cells, and possibly block interferon signaling; iii) In the absence of VP8 we observed reduced numbers of mature virions, so we will assess the role of VP8 in virus assembly; iv) As VP8 is phosphorylated, we will define the impact of VP8 phosphorylation on its functional properties. By characterizing tegument proteins of BHV-1, we will gain insight into the replication and morphogenesis of a complex virus, which will have implications for prevention and control of herpesvirus infections. The availability of cattle as the natural host for in vivo testing provides an advantage for the development of vaccines and therapeutics, not only for BHV-1 but also for related herpesviruses, which will benefit human health as well as the livestock industry in Canada.

疱疹病毒由四个同心区室组成，即围绕 DNA 的核蛋白核心、衣壳、外皮和包膜。外皮含有至少 15 种蛋白质，这些蛋白质在病毒感染中发挥着重要作用，因为它们首先与细胞内环境相互作用，并在后期结合产生新的病毒粒子。该计划的长期目标是对牛疱疹病毒 1 (BHV-1)（牛呼吸道疾病的主要病原体之一）的皮膜蛋白进行功能表征。该提案的具体目标是进一步表征 VP8 的功能，VP8 是 BHV-1 中的主要外皮蛋白。其他外皮蛋白，特别是那些与 VP8 相互作用的蛋白，将按照类似的方法进行研究。缺乏 VP8 的突变病毒在体外受到损害，在体内无毒力，这表明除了结构作用外，VP8 还具有重要的早期功能，可能调节先天免疫反应。我们的假设是，VP8 在建立有效病毒复制的条件和促进病毒组装方面都发挥着作用。我们将进行如下测试： i) 由于 VP8 具有 RNA 结合活性，我们将检查 VP8 对病毒 mRNA 转运的早期影响； ii) 有趣的是，VP8 与 DNA 损伤结合蛋白-1 和 culllin4A 相互作用，因此我们将确定 VP8 是否劫持 cullin-4A-DDB1 E3 泛素连接酶来控制受感染细胞中的蛋白质降解，并可能阻断干扰素信号传导； iii) 在没有 VP8 的情况下，我们观察到成熟病毒体数量减少，因此我们将评估 VP8 在病毒组装中的作用； iv) 由于 VP8 被磷酸化，我们将定义 VP8 磷酸化对其功能特性的影响。通过表征 BHV-1 的外被蛋白，我们将深入了解复杂病毒的复制和形态发生，这将对疱疹病毒感染的预防和控制产生影响。牛作为体内测试的天然宿主为疫苗和治疗方法的开发提供了优势，不仅针对 BHV-1，而且针对相关的疱疹病毒，这将有利于人类健康以及加拿大的畜牧业。