Disabling of the Anaphase Promoting Complex by Human Cytomegalovirus

人巨细胞病毒使后期促进复合物失活

基本信息

批准号：
7908801
负责人：
DEBORAH Hye SPECTOR
金额：
$ 18.47万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-12 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7908801
关键词：
Affect Bacterial Artificial Chromosomes Binding Cell Communication Cell Cycle Cell Cycle Arrest Cell Line Cells Cellular Assay Complement Complementary DNA Complex Congenital Abnormality Cyclin-Dependent Kinases Cytomegalovirus Cytomegalovirus Infections DNA biosynthesis Degradation Pathway Disabled Persons Electroporation Fibroblasts Geminin Gene Expression Genes Goals Grant Human Immunocompromised Host Individual Infection Laboratories Libraries Measures Molecular Molecular and Cellular Biology Morbidity - disease rate Mutation Nuclear Protein Nuclear Proteins Open Reading Frames Pathogenesis Phase Plaque Assay Production Proteins Protocols documentation Red nucleus structure Regulatory Pathway Rest Risk Factors Sampling Signal Transduction Time Ubiquitin Vascular Diseases Viral Viral Genes Viral Pathogenesis Viral Proteins Virus anaphase-promoting complex base cDNA Library gene function immunosuppressed mortality multicatalytic endopeptidase complex mutant protein function public health relevance reconstitution research study ubiquitin-protein ligase viral DNA

项目摘要

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) is the leading viral cause of birth defects, causes significant morbidity and mortality in immunosuppressed individuals, and has been proposed as a risk factor in vascular disease. This virus has evolved multiple mechanisms to usurp cellular signaling and regulatory pathways to facilitate its replication. A key question has been how the virus activates the cell to a state that is optimal for DNA replication, and at the same time alters the levels and activity of selected cellular proteins so that viral replication proceeds at the expense of the host. We have found that one mechanism by which HCMV exploits the host cell involves the ubiquitin-proteasome degradation pathway. A specific target of the virus is the multisubunit Anaphase Promoting Complex (APC) E3 ubiquitin ligase. During the early phase of the infection, this complex is disabled, allowing stabilization and accumulation of its substrates. The effects on the APC include degradation of the APC4 and APC5 subunits, hyperphosphorylation of Cdh1, a loss of binding of both the APC1 subunit and Cdh1 to subcomplex core of the APC (APC subunits 3, 6, 7, 8) and relocalization of the APC subunits. We propose to determine the viral gene(s) that are responsible for this dysregulation of the APC and have developed a high throughput cellular assay for this purpose. There are two complementary approaches that we will take. The first utilizes a library of cDNA constructs corresponding to all of the HCMV open reading frames (ORFs), and the second utilizes a library of HCMV Bacterial Artificial Chromosomes (BACs) that have mutations in each of the ORFs. These results will provide the basis for more in-depth studies on the molecular and cellular mechanisms involved and the functions of these genes with respect to viral replication and pathogenesis. PUBLIC HEALTH RELEVANCE: Human Cytomegalovirus (HCMV) is the major viral cause of birth defects, poses a serious problem for immunocompromised individuals, and has been proposed to be a risk factor for vascular disease. The serious problems associated with HCMV infections have provided a major impetus for understanding the molecular and cellular biology of the virus and the regulatory pathways governing its replication and interactions with the host.

描述（由申请人提供）：人类巨细胞病毒（HCMV）是导致出生缺陷的主要病毒原因，在免疫抑制个体中引起显着的发病率和死亡率，并且已被提议作为血管疾病的危险因素。这种病毒已经进化出多种机制来篡夺细胞信号传导和调节途径以促进其复制。一个关键问题是病毒如何将细胞激活到最有利于DNA复制的状态，同时改变选定细胞蛋白的水平和活性，从而使病毒复制以牺牲宿主为代价进行。我们发现 HCMV 利用宿主细胞的一种机制涉及泛素-蛋白酶体降解途径。该病毒的一个特定靶标是多亚基后期促进复合物 (APC) E3 泛素连接酶。在感染的早期阶段，该复合物被禁用，从而使其底物稳定并积累。对 APC 的影响包括 APC4 和 APC5 亚基的降解、Cdh1 的过度磷酸化、APC1 亚基和 Cdh1 与 APC 亚基复合体核心（APC 亚基 3、6、7、8）的结合丧失以及 APC 亚基的重新定位。 APC 亚基。我们建议确定导致 APC 失调的病毒基因，并为此开发了一种高通量细胞检测方法。我们将采取两种互补的方法。第一个利用与所有 HCMV 开放阅读框 (ORF) 相对应的 cDNA 构建体文库，第二个利用每个 ORF 中都有突变的 HCMV 细菌人工染色体 (BAC) 文库。这些结果将为更深入地研究所涉及的分子和细胞机制以及这些基因在病毒复制和发病机制方面的功能提供基础。公共卫生相关性：人类巨细胞病毒 (HCMV) 是导致出生缺陷的主要病毒原因，给免疫功能低下的个体带来严重问题，并被认为是血管疾病的危险因素。与 HCMV 感染相关的严重问题为了解病毒的分子和细胞生物学以及控制其复制和与宿主相互作用的调控途径提供了主要动力。