Elucidating the mechanism of UL138 mediated suppression of lytic phase genes during human cytomegalovirus latency.

阐明 UL138 介导的人巨细胞病毒潜伏期裂解期基因抑制的机制。

• 批准号: 9753697
• 负责人: Christopher Gelbmann
• 金额: $ 4.95万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9753697
• 关键词: ABCC1 gene; AIDS/HIV problem; Adult; Architecture; Cell Nucleus; Cell surface; Clinical; Cytomegalovirus; Disease; Early Promoters; Epidermal Growth Factor Receptor; Fibroblasts; Genes; Goals; Golgi Apparatus; Histone Deacetylase Inhibitor; Human; Immune; Immune system; Immunocompromised Host; Infection; Lead; Life; Lysine; Lysosomes; Lytic Phase; Maintenance; Mediating; Modeling; Molecular; Pathway interactions; Patients; Play; Population; Proteins; Repression; Research; Role; Serial Passage; Signal Transduction; Sorting - Cell Movement; Testing; Transmembrane Domain; Transplantation; Tumor Necrosis Factor Receptor; Valproic Acid; Viral Proteins; Virus; cellular targeting; combat; histone demethylase; histone modification; human pathogen; latent infection; pathogen; prevent; recruit; success; trafficking

Project summary Human cytomegalovirus (HCMV) is a major human pathogen infecting 60-90% of the population. Its ability to establish latency and persist for the life of the host contributes to its widespread success as a pathogen. While infection in healthy adults is relatively asymptomatic HCMV can cause severe disease in immunocompromised patients such as transplant and HIV/AIDS patients and those with undeveloped immune systems. Importantly disease can be caused by virus reactivating after decades of latency when immune defenses are compromised. The viral protein UL138 is required to establish and maintain latency and is retained in all clinical isolates but lost upon serial passage in fibroblasts (25, 26, 28). UL138’s critical role during latency makes it an ideal target to combat latent infection. When expressed from a latency defective HCMV strain AD169, UL138 prevents histone deacetylase (HDAC) inhibitor valproic acid (VPA) induced expression of the viral protein IE1 from the major immediate early promoter (MIEP); blocking the first step of a lytic infection (27). This critical checkpoint determines whether HCMV will undergo a lytic or latent infection. Initial studies have found UL138 prevents lysine demethylases (KDMs) from removing repressive histone modifications on the MIEP (27). Intriguingly, UL138 localizes to the Golgi apparatus and has not been found in the nucleus where its effect on the MIEP takes place. The primary goal of this study is to identify how UL138 prevents demethylases from being recruited to the MIEP allowing for the establishment of latency. A clearer understanding of how UL138 helps HCMV establish and maintain latency is needed to develop treatments to combat latent HCMV and to clear the latent reservoir. Through our studies possible cellular targets to modulate HCMV latency will be discovered. This research is driven by the hypothesis that UL138 alters cellular trafficking of an unidentified protein(s) preventing them from recruiting KDMs to the MIEP. Supporting this hypothesis UL138 contains four Golgi sorting motifs and alters the subcellular localization of several proteins (33, 34, 35, 37). The requirement for these sorting motifs and cellular sorting machinery for UL138 mediated suppression of the MIEP will be tested and the proteins UL138 must interact with to silence the MIEP will be identified.

项目总结 人巨细胞病毒(HCMV)是一种主要的人类病原体，可感染60-90%的人群。它有能力 建立潜伏期并在寄主的一生中持续存在，有助于其作为病原体的广泛成功。而当 健康成人感染相对无症状的人巨细胞病毒可引起严重疾病中的免疫功能低下 移植患者和艾滋病毒/艾滋病患者以及免疫系统不发达的患者。重要的是 疾病可能是由病毒在数十年的延迟后重新激活引起的，此时免疫防御是 妥协了。病毒蛋白UL138是建立和维持潜伏期所必需的，并保留在所有 临床分离株，但在成纤维细胞连续传代时丢失(25，26，28)。UL138‘S在潜伏期的关键作用 使其成为对抗潜伏感染的理想目标。当从潜伏缺陷的HCMV株中表达时 AD169，UL138拮抗组蛋白脱乙酰酶(HDAC)抑制剂丙戊酸(VPA)诱导的 来自主要直接早期启动子(MIEP)的病毒蛋白IE1；阻断裂解感染的第一步(27)。 这一关键关卡决定了人巨细胞病毒是裂解性感染还是潜伏性感染。初步研究表明 发现UL138可阻止赖氨酸去甲基酶(KDM)去除抑制性组蛋白修饰 Miep(27)。有趣的是，UL138定位于高尔基体，而没有在细胞核中发现 它对MIEP的影响发生了。 这项研究的主要目标是确定UL138如何阻止去甲基酶被招募到 MIEP允许建立等待时间。更清楚地了解UL138如何帮助HCMV 需要建立和维持潜伏期，以开发对抗潜伏性HCMV和清除潜伏期的治疗方法 水库。通过我们的研究，可能发现了调节HCMV潜伏期的细胞靶点。这 研究是由UL138改变一种未知蛋白质的细胞运输这一假设推动的(S) 阻止他们招募KDM加入MIEP。支持这一假设的UL138包含四个高尔基 对几种蛋白质(33、34、35、37)进行分类并改变其亚细胞定位。对以下方面的要求 这些分选基序和用于UL138介导的MIEP抑制的细胞分选机械将被测试 并将确定UL138必须与之相互作用以使MIEP沉默的蛋白质。