Analysis of the mechanism of HCMV cytoplasmic envelopment

HCMV胞质包膜机制分析

基本信息

批准号：
10659275
负责人：
Christopher C Norbury
金额：
$ 48.76万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-17 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10659275
关键词：
Address Antiviral Agents Biological Assay Capsid Cell Nucleus Clinical Complex Cytomegalovirus Cytoplasm DNA Packaging DNA biosynthesis Data Disease Early Endosome Endosomes Event Fluorescence Geographic Locations Human Image Immunocompromised Host Individual Infection Intervention Investigation Knowledge Learning Life Cycle Stages Link Mass Spectrum Analysis Mediating Membrane Molecular Molecular Motors Neck Newborn Infant Organelles Predisposition Pregnant Women Process Protein Analysis Proteins Publishing Role Site Structural Models Therapeutic Therapeutic immunosuppression Viral Viral Proteins Virion Virus Virus Assembly Virus Replication Work aging population antiviral drug development clinically relevant experimental study improved insight interest novel novel therapeutic intervention resistance mutation screening socioeconomics targeted treatment trafficking viral fitness

项目摘要

Although largely asymptomatic, human cytomegalovirus (HCMV) can cause severe and even fatal disease in a subset of susceptible individuals. While great progress has been made in understanding essential stages of HCMV replication, a detailed description of many of these processes is lacking. Of particular interest in this proposal is the maturation of HCMV virions, namely tegument acquisition and cytoplasmic envelopment. To provide a molecular description of these events, it is important to identify the factors involved, both viral and cellular. This proposal will focus on two viral proteins, UL88 and UL71. We have previously published a role for UL88 in packaging a subset of tegument proteins into the virion tegument layer and the absence of UL88 decreases viral fitness. Previous work has identified UL71 as an envelopment factor that potentially mediates membrane scission, as viruses lacking UL71 are trapped at various stages of budding. The experiments in this proposal will seek to elucidate the molecular details of how UL88 and UL71 drive tegument acquisition and envelopment, respectively. This includes a detailed analysis of functional regions on each protein as well as an investigation into additional factors that potentially contribute to each process. We will investigate the role of EEA1+ endosomes in tegument acquisition and for the membrane scission factor DNM1 in cytoplasmic envelopment. This proposal will utilize a novel fluorescence-based envelopment to identify additional cellular proteins that participate in envelopment. Taken together, these studies will further our understanding of the molecular events that drive the late stages of HCMV maturation and identify novel ways in which HCMV assembly can be targeted as a potential intervention.

尽管大部分无症状，但人类巨细胞病毒 (HCMV) 可以在一部分易感人群中引起严重甚至致命的疾病。虽然在了解 HCMV 复制的基本阶段方面已经取得了巨大进展，但仍缺乏对其中许多过程的详细描述。该提议特别令人感兴趣的是 HCMV 病毒颗粒的成熟，即外皮获得和细胞质包膜。为了提供这些事件的分子描述，重要的是确定所涉及的病毒和细胞因素。该提案将重点关注两种病毒蛋白：UL88 和 UL71。我们之前发表过 UL88 在将一部分外皮蛋白包装到病毒体外皮层中的作用，并且 UL88 的缺失会降低病毒的适应性。先前的研究已确定 UL71 是一种可能介导膜断裂的包膜因子，因为缺乏 UL71 的病毒会在出芽的各个阶段被捕获。本提案中的实验将寻求阐明 UL88 和 UL71 如何分别驱动外皮获取和包封的分子细节。这包括对每种蛋白质功能区域的详细分析，以及对可能影响每个过程的其他因素的调查。我们将研究 EEA1+ 内体在外皮获取中的作用以及膜断裂因子 DNM1 在细胞质包膜中的作用。该提案将利用一种新型的基于荧光的包膜来识别参与包膜的其他细胞蛋白。总而言之，这些研究将进一步加深我们对驱动 HCMV 成熟后期的分子事件的理解，并确定将 HCMV 组装作为潜在干预措施的新方法。