PPiSeq: High-Throughput Protein-Protein Interaction Sequencing

PPiSeq：高通量蛋白质-蛋白质相互作用测序

• 批准号: 10294207
• 负责人: SASHA F LEVY
• 金额: $ 41.69万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-16 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10294207
• 关键词: DHFR gene; Open Reading Frames; protein protein interaction

Project Summary/Abstract Emerging and endemic viral pathogens are a persistent threat to human health, the global economy, and national readiness. Viral proteins interact with host proteins to hijack host cells and replicate transmissible viral particles. Human-viral and viral-viral protein-protein interactions (PPls) have been comprehensively characterized for a limited set of viruses, identifying a "PPI profile" for each virus screened. However, these efforts have characterized only a small fraction of the known viruses. A complete viral-human PPI map would be an invaluable resource, enabling analyses of how often interacting viral proteins converge on common human protein targets, cellular functions, or pathways, and which of these interactions are associated with transmissibility or virulence. Combining the viral-human PPI map with human population genetic analyses will enable characterization of the molecular mechanisms underlying extant and ancient epidemics and how these PPIs drive much of human adaptation. In addition, PPI profiles of human viruses could be used to aid screening of animal reservoirs to identify potential threats before a zoonotic spillover occurs. Despite its great promise, characterization of the viral-human PPI map remains a challenge given the throughput of current viral-human PPI screening assays. Current high-throughput assays also lack a quantitative output, meaning that the emergent human-viral PPI map, or viral-viral PPI maps, would be difficult to exploit for important downstream variant scanning or drug screening applications. Here we will use a quantitative sequencing-based protein-protein interaction assay platform to screen for PPls between -24 million viral-human or viral-viral protein pairs. We will further develop this technology into a massively parallel drug screening platform and use it to screen >3 million drug-PPI combinations for small-molecule compounds that promote or antagonize a PPI. The viral-human PPI and drug-PPI maps developed here will be an invaluable resource for a broad research community. In addition, this work will establish new massively parallel and quantitative PPI and drug-PPI screening technologies that will scale with advances in gene synthesis, mutagenesis and sequencing, enabling parallel screening of tens of thousands of gene and gene variants of extant, emerging, and potentially zoonotic viruses.

项目摘要/摘要 新兴和地方性的病毒病原体是对人类健康，全球经济和民族准备的持续威胁。病毒蛋白与宿主蛋白与劫持宿主细胞相互作用并复制可传播的病毒颗粒。人类病毒和病毒 - 病毒蛋白 - 蛋白质相互作用（PPLS）已针对有限的病毒进行了全面特征，为每个筛查的病毒识别一个“ PPI谱”。但是，这些努力仅特征了一小部分已知病毒。完整的病毒人类PPI图将是一个宝贵的资源，可以分析相互作用的病毒蛋白在常见的人类蛋白质靶标，细胞功能或途径上收敛，并且这些相互作用中的哪些相互作用与传播性或病毒性有关。将病毒 - 人类PPI图与人类种群遗传分析相结合，可以表征现存和古代流行病的分子机制，以及这些PPI如何驱动大部分人类适应性。此外，可以使用人类病毒的PPI特征来帮助筛查动物储层，以在发生人畜共患病之前识别潜在的威胁。尽管有巨大的希望，但鉴于当前病毒 - 人类PPI筛选测定法的吞吐量，病毒人类PPI图的表征仍然是一个挑战。当前的高通量测定法也缺乏定量输出，这意味着很难利用重要的下游变体扫描或药物筛查应用来利用新兴的人类病毒PPI图或病毒 - 病毒PPI图。在这里，我们将使用基于定量测序的蛋白质 - 蛋白质相互作用测定平台来筛选-2400万个病毒 - 人类或病毒 - 病毒蛋白对之间的PPL。我们将进一步将该技术开发到一个大规模平行的药物筛选平台中，并将其用于筛选300万个药物-PPI组合，以促进或拮抗PPI的小分子化合物。这里开发的病毒性PPI和毒品PPI地图将是广泛的研究社区的宝贵资源。此外，这项工作将建立新的平行和定量PPI和药物ppi筛选技术，这些技术将随着基因合成，诱变和测序的进步而扩展，从而可以平行筛选出现，出现的，出现的，以及潜在的动物病毒病毒的基因和基因变异。