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.

尽管在很大程度上是无症状的，但人类巨细胞病毒（HCMV）可能会在易感人群的一部分中引起严重甚至致命的疾病。尽管在理解HCMV复制的基本阶段方面取得了巨大进展，但缺乏对许多这些过程的详细描述。在这一建议中特别感兴趣的是HCMV病毒体的成熟，即收购和细胞质包膜。为了提供这些事件的分子描述，重要的是要确定涉及的因素，包括病毒和细胞。该提案将重点介绍两种病毒蛋白UL88和UL71。我们以前曾在将Tegument蛋白的子集包装到病毒座层中，并发表了UL88的作用，并且不存在UL88会降低病毒适应性。先前的工作已经将UL71确定为可能介导膜分裂的包络因子，因为缺乏UL71的病毒被困在各个萌芽阶段。该提案中的实验将旨在阐明UL88和UL71分别驱动tegument的获取和信封的分子细节。这包括对每种蛋白质功能区域的详细分析，以及对潜在有助于每个过程的其他因素的研究。我们将研究EEA1+内体在TEGUMEMENT获得的作用以及在细胞质包膜中膜分裂因子DNM1的作用。该建议将利用一种新型的基于荧光的信封来识别参与包膜的其他细胞蛋白。综上所述，这些研究将进一步理解驱动HCMV成熟后期阶段的分子事件，并确定可以将HCMV组装作为潜在干预措施的新颖方式。