HHV-8 vIRF interactions in the context of infection

HHV-8 vIRF 在感染情况下的相互作用

基本信息

批准号：
9085244
负责人：
John Nicholas
金额：
$ 21.14万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9085244
关键词：
Address Affinity Antiviral Agents Apoptosis Apoptotic BCL2 gene Binding Biological Biology Cell Cycle Arrest Cell Survival Cells Clinical Cytostatics Data Development EP300 gene Endothelial Cells Evaluation Family Genes Genetic Transcription Goals Health Homologous Gene Human Herpesvirus 8 Immune Immune response Individual Infection Interferons Kaposi Sarcoma Knowledge Latent Virus Life Cycle Stages Lymphoma Macaca mulatta Malignant - descriptor Malignant Neoplasms Mass Spectrum Analysis Mediating Multicentric Angiofollicular Lymphoid Hyperplasia Natural Immunity Pathogenesis Pathway interactions Peptides Phase Precipitation Production Proteins Reagent Reporting Research Rhadinovirus Role Specific qualifier value Stress Surveys TP53 gene Therapeutic Therapeutic Effect Time Transcription Coactivator Tretinoin Viral Viral Proteins Virus Virus Diseases Virus Latency Work ataxia telangiectasia mutated protein base cellular targeting in vivo inhibitor/antagonist lytic replication member novel overexpression prevent primary effusion lymphoma response viral interferon regulatory factor viral interferon regulatory factor-1

项目摘要

DESCRIPTION (provided by applicant): Human herpesvirus 8 (HHV-8) specifies four viral interferon regulatory factor homologues (vIRFs) that represent a novel set of virus-encoded proteins, so-far identified only in HHV-8 and closely related rhesus rhadinovirus. The HHV-8 vIRFs function to inhibit cellular IRF activities and other components of cellular defense pathways that promote cell cycle arrest and apoptosis in response to virus infection. Two of the vIRFs, vIRF-1 and vIRF-3, have been demonstrated to contribute significantly to either virus productive replication (in endothelial cells) or to latently infected cell viability [in primary efusion lymphoma (PEL) cells]; each vIRF is expressed during both latent (PEL) and lytic replication. Both phases of the virus life cycle are important for HHV-8 malignant pathogenesis: Kaposi's sarcoma, PEL, and multicentric Castleman's disease. Known targeted cellular proteins of vIRF-1 include IRFs, p300/CBP transcriptional co-activators required for IRF-mediated responses, IFN/retinoic acid-induced GRIM19, p53, p53-activating ATM kinase, and certain BH3-only (pro-apoptotic) members of the Bcl-2 family; vIRF-3 targets IRFs, p300/CBP, and p53. However, the full repertoire of inhibitory interactions of these pleiotropic proteins is far from clear and the interactions have not been adequately surveyed in the context of latent and lytic replication. Whilst we have determined that vIRF-1 interactions with BH3-only proteins are critical for successful productive replication in endothelial cells, the roles of other vIRF-1 interactions in lytic replication in these and other cells and the contributions of vIRF-1 and its interactions to PEL latency remain unknown. Similarly, the roles of vIRF-3 and its interactions in productive replication and of specific vIRF-3 interactions in latency remain to be determined. To address these issues, we propose to: (1) undertake affinity precipitations of vIRFs 1 and 3 from latently infected PEL and lytically infected PEL and endothelial cells followed by mass spectrometry analysis of co-precipitated cellular proteins; (2) use suppression, overexpression, and binding-competition analyses to identify the contributions of vIRFs 1 and 3, their cellular targets, and particular vIRF-protein interactions to latently infected cell viability and productive replication and to develop inhibitory agents. The project comprises a broadly-visioned but conceptually and experimentally focused analysis of unique viral innate immune evasion proteins whose functional and mechanistic contributions to virus biology are presently unclear. As these proteins are centrally important to both latency and productive replication, information generated from this study could provide the basis for development of novel antiviral and clinical therapies.

描述（由申请人提供）：人类疱疹病毒 8 (HHV-8) 指定了四种病毒干扰素调节因子同源物 (vIRF)，它们代表一组新型病毒编码蛋白，迄今为止仅在 HHV-8 和密切相关的恒河猴病毒中发现HHV-8 vIRF 的功能是抑制细胞 IRF 活性和细胞防御途径的其他成分，从而促进细胞周期停滞和细胞凋亡以应对病毒感染。 vIRF、vIRF-1 和 vIRF-3 已被证明对病毒的有效复制（在内皮细胞中）或潜伏感染的细胞活力（在原发性渗出性淋巴瘤 (PEL) 细胞中）有显着贡献；每个 vIRF 在潜伏期间均表达； (PEL) 和裂解复制对于 HHV-8 恶性发病机制都很重要：卡波西肉瘤、PEL 和多中心 Castleman 肉瘤。 vIRF-1 的已知靶细胞蛋白包括 IRF、IRF 介导的反应所需的 p300/CBP 转录共激活因子、IFN/视黄酸诱导的 GRIM19、p53、p53 激活 ATM 激酶和某些 BH3-only（pro） -凋亡）Bcl-2 家族成员；vIRF-3 靶向 IRF、p300/CBP 和 p53。这些多效性蛋白的抑制性相互作用的全部内容尚不清楚，并且在潜伏和裂解复制的背景下尚未充分研究这些相互作用，尽管我们已经确定 vIRF-1 与 BH3-only 蛋白的相互作用对于成功进行高效复制至关重要。内皮细胞中，其他 vIRF-1 相互作用在这些细胞和其他细胞的裂解复制中的作用以及 vIRF-1 及其相互作用对 PEL 潜伏期的贡献仍然未知。 vIRF-3 及其在生产性复制中的相互作用以及潜伏期中特定 vIRF-3 的相互作用仍有待确定。为了解决这些问题，我们建议：（1）对潜伏感染的 PEL 和裂解感染的 vIRF 1 和 3 进行亲和沉淀。 PEL 和内皮细胞，然后对共沉淀的细胞蛋白进行质谱分析；(2) 使用抑制、过度表达和结合竞争分析来确定 vIRF 的贡献 1 3、它们的细胞靶标，以及特定的 vIRF 蛋白与潜伏感染细胞活力和高效复制的相互作用该项目包括对独特的病毒先天免疫逃避蛋白进行广泛但在概念和实验上集中的分析，这些蛋白对病毒生物学的功能和机制贡献目前尚不清楚，因为这些蛋白对于潜伏和有效复制都至关重要。，这项研究产生的信息可以为开发新型抗病毒和临床疗法提供基础。