Overcoming host Genetic Redundancy and Pathogen Subversion to Define new host-viral Interfaces

克服宿主遗传冗余和病原体颠覆，定义新的宿主-病毒界面

• 批准号: 10654047
• 负责人: Robert C. Orchard
• 金额: $ 40.51万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-18 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10654047
• 关键词: Area; Attenuated; Binding; Biochemical; Biology; Bypass; Cell physiology; Communicable Diseases; Complex; Complication; Conflict (Psychology); Disparate; Epigenetic Process; Family; Genes; Genetic; Genetic Screening; Genome; Histone Acetylation; Host Defense; Immune Evasion; Infection; Influenza; Integration Host Factors; Knowledge; Ligands; Maps; Molecular; Mus; Mutation; Norovirus; Orphan; Pathway interactions; Physiological; Protein Family; Proteins; System; Viral; Viral Genes; Virulent; Virus; Virus Replication; comparative; gain of function; insight; pathogen; programs; protein function; receptor; screening; tool

Project Summary/Abstract Viruses have evolved sophisticated mechanisms to hijack host cellular machinery. For their part, hosts have developed their own intricate defense systems. Defining how these opposing strategies have co-evolved increases our understanding of infectious diseases and provides new opportunities to discover unexplored areas of biology. Our understanding of host cellular defense systems has been limited due to genetic redundancy in host genomes. A further complication is that successful pathogens are able to inactivate host antiviral networks during infection. We have recently established a new genetic screening platform that overcomes these roadblocks. First, we bypass genetic redundancy in host genomes through a gain of function screen that identifies antiviral genes. Second, we use both virulent and attenuated strains of viruses in our screening pipeline. Importantly, these attenuated viruses have deletions or mutations in critical, yet poorly understood, immune evasion proteins that antagonize host antiviral restriction factors. We predict that the avirulent strains of these viruses will become sensitive to restriction factors that have otherwise defied molecular identification. Using this strategy, one project will be to identify ancient antiviral genes that reveal unappreciated areas of host- pathogen conflict. For example, we are unmasking how the influenza immune evasion protein, NS1 orchestrates a complex, multifaceted rewiring of host antiviral networks using comparative screens with wild-type and NS1-deficient viruses. We are extending these studies to other virulent-attenuated virus pairings as well. Another project takes newly identified antiviral molecules that inhibit disparate virus families and define the molecular mechanism underlying viral inhibition. Here, we focus on the JADE family of proteins that regulate histone acetylation. We aim to uncover functional redundancy in the JADE family and to determine if it assembles a concerted antiviral epigenetic program. Another project is to leverage newly identified host-pathogen conflicts to discover the physiological functions of host- proteins that are poorly understood. In this regard, we are focusing on the CD300 family of receptor proteins. CD300 genes are undergoing rapid positive selection, yet their physiological function remains largely unexplored. We previously demonstrated that CD300lf is a protein receptor for murine norovirus. We will employ the biochemical, genetic, and cellular tools we developed for studying norovirus-CD300 interactions to define the physiological ligands of CD300 receptors. Determining the ligands for these orphan receptors will provide functional insight into these rapidly evolving proteins. Taken together, we plot a road map for discovering new host-pathogen interfaces, defining their molecular interactions during host defense, and how this relates to the physiological functions of these proteins and pathways.

项目概要/摘要 病毒已经进化出复杂的机制来劫持宿主细胞机器。就他们而言， 主机开发了自己复杂的防御系统。定义这些相反的策略如何 共同进化增加了我们对传染病的了解，并提供了新的机会 发现生物学中未探索的领域。我们对宿主细胞防御系统的理解是 由于宿主基因组中的遗传冗余而受到限制。进一步的复杂化是成功的病原体 能够在感染期间灭活宿主的抗病毒网络。我们最近建立了一个新的 克服这些障碍的基因筛查平台。首先，我们绕过遗传冗余 通过识别抗病毒基因的功能获得筛选来识别宿主基因组。其次，我们同时使用 我们筛选管道中的强毒株和减毒株。重要的是，这些减弱了 病毒在关键但仍知之甚少的免疫逃避蛋白中存在缺失或突变，这些蛋白 拮抗宿主抗病毒限制因素。我们预测这些病毒的无毒株将 对限制因素变得敏感，否则无法进行分子鉴定。使用这个 作为一项战略，一个项目将是识别古老的抗病毒基因，这些基因揭示了宿主未被认识的区域 病原体冲突。例如，我们正在揭示流感免疫逃避蛋白 NS1 如何 使用比较屏幕来协调宿主抗病毒网络的复杂、多方面的重新布线 野生型和 NS1 缺陷型病毒。我们正在将这些研究扩展到其他毒力减毒病毒 配对也是如此。另一个项目采用新发现的抑制不同病毒的抗病毒分子 家族并定义病毒抑制的分子机制。在这里，我们重点关注JADE 调节组蛋白乙酰化的蛋白质家族。我们的目标是发现功能冗余 JADE家族并确定它是否组装了一个协调一致的抗病毒表观遗传程序。另一个项目 是利用新发现的宿主-病原体冲突来发现宿主-病原体的生理功能 人们对蛋白质知之甚少。在这方面，我们重点关注CD300受体家族 蛋白质。 CD300基因正在经历快速正选择，但其生理功能仍然存在 很大程度上尚未探索。我们之前证明 CD300lf 是鼠诺如病毒的蛋白质受体。 我们将利用我们为研究诺如病毒-CD300而开发的生化、遗传和细胞工具 相互作用来定义 CD300 受体的生理配体。确定这些的配体 孤儿受体将为这些快速进化的蛋白质提供功能洞察。综合起来，我们 绘制发现新宿主-病原体界面的路线图，定义它们的分子相互作用 在宿主防御过程中，以及这与这些蛋白质和途径的生理功能有何关系。