Cytomegalovirus UL80 Proteins:Interactions and Modifications that Impact Function

巨细胞病毒 UL80 蛋白：影响功能的相互作用和修饰

• 批准号: 8263745
• 负责人: D Wade Gibson
• 金额: $ 20.5万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-15 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8263745
• 关键词: Affect; Area; Automobile Driving; Binding; Biological; Biological Assay; Biological Testing; Biology; Capsid; Capsid Proteins; Cell Nucleus; Cells; Coiled-Coil Domain; Cysteine; Cytomegalovirus; DNA Packaging; Dissociation; Drug Delivery Systems; Genes; Glycogen Synthase Kinase 3; Goals; Herpesviridae; Homologous Protein; Immune system; In Vitro; Infection; Lead; MAP Kinase Gene; Methods; Modeling; Modification; Molecular; Nuclear; Nuclear Localization Signal; Open Reading Frames; Peptide Hydrolases; Phenotype; Phosphorylation; Phosphorylation Site; Pilot Projects; Play; Process; Protein Precursors; Proteins; Research; Role; Sedimentation process; Simplexvirus; Site; Solubility; Testing; Therapeutic; Two-Hybrid System Techniques; Virus; Virus Replication; Work; base; mutant; novel strategies; protein protein interaction; protein structure function; public health relevance; research study; success; vector; viral DNA

DESCRIPTION (provided by applicant): We propose to answer questions about molecular interactions guiding the earliest steps in formation of infectious cytomegalovirus. Our focus is on two genetically related proteins encoded by open reading frame UL80a, called the assembly protein precursor (pAP) and the capsid maturational protease precursor (pPR). Together these proteins interact with themselves and with other proteins to coordinate assembly and maturation of the nascent procapsid shell - a process essential to produce infectious virus. We have recently overcome a solubility problem that complicates working with the purified proteins in vitro, and will apply this new approach to pursue leads and test models based on encouraging results from pilot studies. We will do this through three Specific Aims: (i) test predicted effects of phosphorylation on key interactions of pAP and pPR; (ii) verify the differential dominance of the "carboxyl coiled-coil domain" in the self-interaction of pAP versus pPR self-interaction, and determine its biological importance and relevance; and (iii) establish whether a CMV-specific second nuclear localization signal (NLS2) in pAP and pPR is required for their interaction with the capsid portal protein (pUL104). Information gained from this work will help define the sequence of changes that drive UL80 protein associations and dissociations during capsid formation and maturation, and contribute to the longer-term goal of developing new strategies for disrupting (e.g., for therapeutics) or exploiting (e.g., gene/drug delivery vectors) herpesvirus assembly. PUBLIC HEALTH RELEVANCE: Cytomegalovirus is a herpes-group virus that is a threat to people with weakened immune systems, including the very young and the very old. This research seeks to understand how two genetically-related proteins guide the earliest steps in forming infectious virus. Information obtained will relate the molecular interactions driving capsid formation will the biological mechanism of virus formation, and will help identify new ways to interfere therapeutically with that process.

描述（由申请人提供）：我们建议回答有关指导传染性巨细胞病毒形成的最早步骤的分子相互作用的问题。我们的重点是由开放阅读框 UL80a 编码的两种遗传相关蛋白质，称为组装蛋白前体 (pAP) 和衣壳成熟蛋白酶前体 (pPR)。这些蛋白质彼此相互作用并与其他蛋白质相互作用，以协调新生衣壳的组装和成熟——这是产生传染性病毒所必需的过程。我们最近克服了溶解度问题，该问题使体外纯化的蛋白质的工作变得复杂，并将根据试点研究的令人鼓舞的结果应用这种新方法来寻找线索和测试模型。我们将通过三个具体目标来实现这一目标：(i) 测试磷酸化对 pAP 和 pPR 关键相互作用的预测影响； (ii)验证“羧基卷曲螺旋结构域”在pAP自相互作用与pPR自相互作用中的差异优势，并确定其生物学重要性和相关性； (iii) 确定 pAP 和 pPR 中的 CMV 特异性第二核定位信号 (NLS2) 是否是它们与衣壳门蛋白 (pUL104) 相互作用所必需的。从这项工作中获得的信息将有助于定义在衣壳形成和成熟过程中驱动 UL80 蛋白结合和解离的变化顺序，并有助于制定新的破坏策略（例如用于治疗）或利用（例如，基因/药物递送载体）疱疹病毒组装。 公共卫生相关性：巨细胞病毒是一种疱疹病毒，对免疫系统较弱的人（包括幼儿和老年人）构成威胁。这项研究旨在了解两种遗传相关的蛋白质如何指导形成传染性病毒的最早步骤。获得的信息将驱动衣壳形成的分子相互作用与病毒形成的生物学机制联系起来，并将有助于确定治疗干扰该过程的新方法。