Investigating the Role of E3 Ubiquitin Ligases in Flavivirus Infection

登革热（DENV）和寨卡病毒（ZIKV）等黄病毒构成了严重的公共卫生挑战，全球每年感染4亿多人。尽管如此，目前还没有有效的疫苗或抗病毒药物来对抗黄病毒，这突出了我们需要提高对其生物学的理解。虽然已知黄病毒劫持宿主泛素化系统以发挥其优势，但催化泛素化的e3连接酶在黄病毒生命周期中的作用受到的关注有限，对正常e3连接酶生物学的了解有限。本项目旨在鉴定和表征宿主细胞的e3连接酶，这些连接酶被选为对黄病毒生命周期有前病毒效应的连接酶。我们将采用无偏倚和基于候选的方法来鉴定寨卡病毒感染中活性改变的e3连接酶。我们将结合遗传、生化和蛋白质组学策略来详细表征这些E3-连接酶在病毒复制中的作用。最后，我们的目标是验证这些e3连接酶介导的过程在蚊子载体中的保守性，以确定人类和媒介宿主的普遍感染机制。从这项研究中获得的见解将进一步加深我们对；健康e3连接酶生物学和人类免疫功能，黄病毒生物学，并可能有助于广谱抗病毒药物的开发。这项工作属于BBSRC更广泛的主题“生物科学对健康的综合理解”，特别是优先领域“整个生命过程的健康老龄化”和“终身健康和福祉”。我们将了解e3连接酶细胞功能的基本方面，黄病毒感染使我们能够确定e3连接酶的关键相互作用和活性，这对调节更广泛的细胞生物学至关重要。在不同的黄病毒种类和不同的宿主物种中鉴定具有保守功能的e3连接酶，也将使我们能够鉴定在细胞过程（如代谢和免疫反应）中起关键作用的核心e3连接酶。总之，这将提高我们对细胞生物学的理解，并确定关键的e3连接酶，这些连接酶对细胞的健康功能很重要，在疾病中特别容易受到功能障碍的影响。

Flaviviruses such as Dengue (DENV) and Zika (ZIKV) pose serious public health challenges, infecting over 400 million people a year globally. Despite this, no effective vaccines or antivirals are currently available to combat flaviviruses, highlighting the need to improve our understanding of their biology. While it is known that flaviviruses hijack the host ubiquitylation system to their advantage, E3-ligases that catalyse ubiquitylation have received limited attention on their role in the flavivirus lifecycle, with limited understanding of normal E3-ligase biology. This project aims to identify and characterise E3-ligases of the host cell that are co-opted to have a proviral effect on the flavivirus lifecycle. We will apply both an unbiased as well as a candidate-based approach to identify E3-ligases with altered activity in ZIKV infection. We will combine genetic, biochemical and proteomics strategies to perform detailed characterization of these E3- ligases in virus replication. Finally, we aim to verify the conservation of these E3-ligase-mediated processes in the mosquito vector to define universal infection mechanisms in both human and the vector host. The insights gained from this study will further our understanding of; healthy E3-ligase biology and immune function in humans, flavivirus biology, and potentially contribute to the development of broad-spectrum antivirals.BBSRC Relevant Areas This work comes under the wider BBSRC theme of 'Bioscience for an integrated understanding of health', specifically the priority areas 'Healthy ageing across the life course' and 'Lifelong health and wellbeing'. We will gain an understanding of the fundamental aspects of E3-ligase cellular function, with flavivirus infection allowing us to identify key interactions and activities of E3-ligases which are crucial to modulating wider cellular biology. Identifying E3-ligases with conserved functions across different flavivirus species and in different host species will also allow us to identify core E3-ligases which are key in cellular processes such as metabolism and the immune response. Together, this will improve our understanding of cellular biology and also identify key E3-ligases which are important in healthy function of the cell and particularly susceptible to dysfunction in disease.