HCMV US28 Signal Transduction by Betaarrestin proteins

Betaarrestin 蛋白的 HCMV US28 信号转导

• 批准号: 7371077
• 负责人: WILLIAM E MILLER
• 金额: $ 24.99万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7371077
• 关键词: Acquired Immunodeficiency Syndrome; Adenylate Cyclase; Affect; Arrestin; Arrestins; Atherosclerosis; Attenuated; Binding; Biochemical; Biological; Biology; Cell Line; Cells; Complex; Congenital Abnormality; Cytomegalovirus; Cytomegalovirus Infections; Data; Development; Disease; Docking; Event; Extracellular Signal Regulated Kinases; Family; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Genes; Genome; Goals; Herpesviridae; Immune system; Individual; Infection; Inositol Phosphates; Ligands; Mediating; Mitogen-Activated Protein Kinases; Molecular; Organ Transplantation; PTK2 gene; Pathogenesis; Pathway interactions; Phospho-Specific Antibodies; Phospholipase; Phospholipase C; Phosphorylation; Phosphotransferases; Play; Prevention; Process; Production; Property; Proteins; Public Health; Reagent; Recombinants; Recruitment Activity; Regulation; Research Personnel; Rodent; Role; Scaffolding Protein; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Signaling Protein; Simplexvirus; Site; Smooth Muscle Myocytes; System; Transplant Recipients; Vascular Diseases; Viral; Viral Physiology; Work; attenuation; base; cell motility; chemokine; desensitization; design; expectation; genetic regulatory protein; human MAPK14 protein; in vivo; inhibitor/antagonist; innovation; insight; interest; knock-down; member; mitogen-activated protein kinase p38; mutant; novel; programs; protein activation; receptor; scaffold

DESCRIPTION (provided by applicant): Human Cytomegalovirus (HCMV) infection is a leading cause of viral associated congenital birth defects and has proven to be a significant public health problem in organ transplant recipients as well as in individuals with AIDS. Accumulating evidence indicates that there may be a relationship between HCMV and the development of vascular disease. The interplay between HCMV and the immune system is a critical factor in the events leading to initiation of disease. Interestingly, HCMV encodes several genes that appear to modulate immune system function. One of these genes, termed US28, is homologous to the mammalian chemokine G-protein coupled receptors (GPCRs). While progress has recently been made in the identification of signaling pathways downstream of US28, very little is known regarding the molecular details proximal to US28 that contribute to the activation of signaling pathways. The objective of this proposal is to identify specific receptor properties and molecular events proximal to US28 that are critical for activation of known signal transduction pathways. We hypothesize that a detailed study of the molecular interactions of betaarrestin proteins with US28 will generate novel insights into the regulation of US28 signaling. The betaarrestins were originally identified in the context of termination of "traditional" G-protein signaling to effectors such as adenylyl cyclases and phospholipases. This termination process, also known as desensitization, involves receptor phosphorylation by GRKs and binding of betarrestin proteins. It is now clear that the betaarrestins simultaneously function as adapter/scaffold molecules to recruit signaling proteins to receptors in order to activate additional "non-traditional" signaling pathways. We have identified a US28 mutant that is unable to interact with betaarrestin and have generated a significant amount of preliminary data regarding signaling from this mutant. We propose to analyze the involvement of betaarrestin proteins in US28 signaling in order to gain a greater understanding of the proximal events leading to activation of US28 signaling pathways. To accomplish this goal, we propose three specific aims. First, we will characterize "traditional" signaling from this US28 mutant in the context of HCMV infection and will analyze how this mutant affects the ability of US28 to stimulate smooth muscle cell migration. Second, we will analyze "non-traditional" signaling pathways from US28 potentially mediated by betaarrestin recruitment to the US28 carboxy terminal regulatory domain. Third, we will analyze US28 signaling in cells deficient for betaarrestin expression in order to specifically define the involvement of betaarrestins. Our preliminary data indicate that US28 engages ?arrestin proteins suggesting that we will be able to make rapid progress toward our goal of defining the role of these important regulatory proteins in US28 signaling. These studies will identify mechanisms regulating US28 mediated signal transduction and will enhance our understanding of how US28 participates in pathogenesis and initiation of HCMV disease.

描述(申请人提供)：人类巨细胞病毒(HCMV)感染是病毒相关先天性出生缺陷的主要原因，已被证明是器官移植接受者和艾滋病患者的一个重大公共卫生问题。越来越多的证据表明，人巨细胞病毒可能与血管疾病的发生发展有关。巨细胞病毒与免疫系统之间的相互作用是导致疾病发生的关键因素。有趣的是，巨细胞病毒编码了几个似乎调节免疫系统功能的基因。其中一个基因被称为US28，与哺乳动物趋化因子G蛋白偶联受体(GPCRs)同源。虽然最近在鉴定US28下游的信号通路方面取得了进展，但对US28附近参与信号通路激活的分子细节知之甚少。这一建议的目的是确定US28附近对激活已知信号转导途径至关重要的特定受体特性和分子事件。我们假设，对Betaarrestin蛋白与US28的分子相互作用的详细研究将对US28信号的调控产生新的见解。Betaarrestins最初是在终止“传统的”G蛋白信号传递给诸如腺苷环化酶和磷脂酶等效应物的背景下被鉴定出来的。这种终止过程，也称为脱敏，涉及GRKs的受体磷酸化和Betarrestin蛋白的结合。现在很清楚的是，Betaarrestins同时起到适配器/支架分子的作用，将信号蛋白招募到受体，以激活额外的“非传统”信号通路。我们已经鉴定出一个US28突变体，它不能与Betaarrestin相互作用，并产生了大量关于来自该突变体的信号的初步数据。我们建议分析Betaarrestin蛋白在US28信号通路中的作用，以便更好地了解导致US28信号通路激活的近端事件。为了实现这一目标，我们提出了三个具体目标。首先，我们将在巨细胞病毒感染的背景下描述来自US28突变体的“传统”信号，并分析该突变体如何影响US28刺激平滑肌细胞迁移的能力。其次，我们将分析从US28到US28羧基末端调节域的“非传统”信号通路，这些信号通路可能由Betarestin募集介导。第三，我们将分析缺乏Betaarrestin表达的细胞中的US28信号，以明确Betaarrestins的参与。 我们的初步数据表明，US28与arrestin蛋白结合，这表明我们将能够朝着确定这些重要调控蛋白在US28信号中的作用的目标快速取得进展。这些研究将确定US28介导的信号转导的调控机制，并将加深我们对US28如何参与巨细胞病毒病的发病和启动的理解。