Mechanisms of ERK activation in gammaherpesvirus

伽马疱疹病毒中 ERK 激活机制

• 批准号: 8119142
• 负责人: Evonne N Woodson
• 金额: $ 2.86万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8119142
• 关键词: AIDS related neoplasm; Acquired Immunodeficiency Syndrome; Address; Biochemical; Biology; Cell Communication; Cell physiology; Cells; DNA biosynthesis; Data; Detection; Developing Countries; Disease Progression; Environment; Enzymes; Epidemic; Genes; Genetic Transcription; Genomics; Goals; Herpesviridae; Herpesviridae Infections; Highly Active Antiretroviral Therapy; Homologous Gene; Human; Human Herpesvirus 8; Immune system; Immunoblotting; Immunoelectron Microscopy; In Vitro; Incidence; Individual; Infection; Kaposi Sarcoma; Kinetics; Laboratories; Life Cycle Stages; Location; Lytic; MAPK3 gene; MEKs; Macaca mulatta; Malignant Neoplasms; Mass Spectrum Analysis; Mitogen-Activated Protein Kinases; Modeling; Pathogenesis; Pathway interactions; Patients; Phase; Phosphorylation; Phosphotransferases; Play; Production; Proteins; Public Health; Rhadinovirus; Role; Small Interfering RNA; Species Specificity; Specificity; Structure; Techniques; Testing; Translations; Viral; Viral Genes; Viral Proteins; Virion; Virus; Virus Assembly; Virus Diseases; Western Blotting; Work; cancer cell; crosslink; design; extracellular; fluorophore; gamma-2 herpesvirus; gammaherpesvirus; in vivo; inhibitor/antagonist; insight; interest; novel; particle; research study; viral DNA

DESCRIPTION (provided by applicant): Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent of three human malignancies, including Kaposi's sarcoma (KS), the most common AIDS-related neoplasm worldwide. While the widespread use of HAART therapy has resulted in a decline in the number of KS cases in the US, KS incidence has increased in developing countries where the AIDS epidemic continues. The long-term objective of this study is to broaden our understanding of the basic biology of gammaherpesviruses, including structure, assembly, and virus-host cell interactions. With the recent finding of ERK1/2 within the purified virion of Rhesus monkey rhadinovirus (RRV), a close homolog of KSHV, we began to speculate the importance of this pathway in viral infection. We hypothesize that the incorporation of these enzymes may represent a snapshot of the cellular environment during viral infection, reflecting a cellular pathway that is highly active in the vicinity of viral replication and assembly and that may play important roles during virus production. Two specific aims are proposed to test this hypothesis. The first is designed to determine the structural aspect of ERK1/2 incorporation. We will biochemically confirm mass spectrometry data, while also determining structural localization, species-specificity, and approximate abundance of these species within the particle. The second is to define the interaction between RRV and the MEK/ERK pathway during de novo infection. We will determine the kinetics of ERK activation, as well as define a mechanism for late phase activation. With the use of viral DNA replication inhibitors, as well as global transcription and translation inhibitors, we will assess role of potential viral gene (s) and cellular processes in activating ERK during RRV infection. Relevance to public health: Viruses, having evolved with the host, have found ways to manipulate the cellular environment for their benefit. This project is designed to provide insight into how gammaherpesviruses interact with and use the MEK/ERK pathway during viral infection. Due to the weakened immune system of AIDS patients, KS treatment is often limited; however, our studies may identify virus and cancer cell specific markers that may become targeted therapies for KS. This approach may enable us to interfere with infection and subsequent disease progression, even in the absence of a healthy immune system.

描述（由申请人提供）：卡波西肉瘤相关疱疹病毒（KSHV）是三种人类恶性肿瘤的病原体，其中包括卡波西肉瘤（KS），全世界最常见的艾滋病相关肿瘤。虽然HAART疗法的广泛使用导致美国KS病例数下降，但在艾滋病流行的发展中国家，KS发病率却有所增加。这项研究的长期目标是扩大我们对伽马疱疹病毒基础生物学的理解，包括结构、组装和病毒-宿主细胞相互作用。随着最近在恒河猴病毒（RRV）（KSHV 的密切同源物）的纯化病毒体中发现 ERK1/2，我们开始推测该途径在病毒感染中的重要性。我们假设这些酶的掺入可能代表了病毒感染期间细胞环境的快照，反映了在病毒复制和组装附近高度活跃的细胞途径，并且可能在病毒产生过程中发挥重要作用。提出了两个具体目标来检验这一假设。第一个旨在确定 ERK1/2 掺入的结构方面。我们将通过生化方法确认质谱数据，同时确定颗粒内这些物种的结构定位、物种特异性和大致丰度。第二个是定义从头感染过程中 RRV 和 MEK/ERK 通路之间的相互作用。我们将确定 ERK 激活的动力学，并定义后期激活的机制。通过使用病毒 DNA 复制抑制剂以及全局转录和翻译抑制剂，我们将评估潜在病毒基因和细胞过程在 RRV 感染期间激活 ERK 中的作用。与公共卫生的相关性：病毒与宿主一起进化，已经找到了操纵细胞环境以使其受益的方法。该项目旨在深入了解伽玛疱疹病毒在病毒感染过程中如何与 MEK/ERK 通路相互作用并使用 MEK/ERK 通路。由于艾滋病患者的免疫系统较弱，KS治疗往往受到限制；然而，我们的研究可能会识别出病毒和癌细胞的特异性标记物，这些标记物可能成为 KS 的靶向疗法。即使在没有健康的免疫系统的情况下，这种方法也可能使我们能够干扰感染和随后的疾病进展。