Structure and Function of Proxvirus Immune Evasion Domains

项目2：痘病毒免疫逃避结构域的结构和功能。在这个项目中，我们将探索由人畜共患正痘病毒牛痘编码的一大类不同序列蛋白质的功能和机制属性。我们假设牛痘中的十种蛋白质共享一个类似的结构支架，我们称之为痘病毒免疫逃避(PIE)结构域。某些PIE蛋白已知会破坏宿主的免疫监视和效应器功能，我们的目标是系统地确定由牛痘Bright ton Red株编码的所有10种PIE蛋白的配体和受体。这些努力将得到结晶学研究和定量结合分析的补充，以确定PIE蛋白质的结构和特异性，以及开发具有改变配体/受体相互作用的变异体的基于结构的工程。我们还将对PLE进行广泛的体外功能研究，在成熟的免疫学检测的背景下评估它们的作用机制。我们的调查也将在病毒感染和宿主防御的更广泛背景下进行考虑。在与横山和Fruh实验室的密切合作下，我们将在选择性PIE蛋白被敲除或其配体/受体结合决定因素改变的情况下评估牛痘的发病机制。拟议的实验将大大改进牛痘的功能注释，并产生关于宿主-病原体相互作用的重要线索，这可能会使科学界和临床界都受益。

Project 2: Structure and function of poxvirus immune evasion domains. In this project we will explore the functional and mechanistic attributes of a large family of sequence diverse proteins encoded by the zoonotic orthopoxvirus cowpox. We hypothesize that ten proteins In cowpox share a similar structural scaffolding that we have termed the poxvirus immune evasion (PIE) domain. Certain PIE proteins are known to sabotage host Immune surveillance and effector functions, and our goal Is to systematically determine the ligands and receptors for all ten PIE proteins encoded by the Brighton Red strain of cowpox. These efforts will be complemented by crystallographic investigations and quantitative binding assays to determine PIE protein structure and specificity, and structure-based engineering to develop variants with altered ligand/receptor Interactions. We will also undertake extensive In vitro functional studies of the PlEs, evaluating their mechanisms of action In the context of well-established Immunological assays. Our investigations will also be considered In the broader context of the battlefield of viral infection and host defense. In close collaboration with the Yokoyama and Fruh labs we will evaluate the pathogenesis of cowpox with either selective PIE proteins knocked out or their ligand/receptor binding determinants altered. The proposed experiments will result In a significantly improved functional annotation of cowpox and yield important clues about host-pathogen Interactions that are likely to benefit both the scientific and clinical communities.