Gene regulatory role of vIRF1 in KSHV infection

vIRF1在KSHV感染中的基因调控作用

• 批准号: 10464138
• 负责人: Seung Jin Jang
• 金额: $ 4.53万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-16 至 2024-08-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10464138
• 关键词: Amino Acids; Antiviral Response; Antiviral Therapy; B-Cell Lymphomas; Binding; Biological; Biological Assay; C-terminal; Cell Communication; Cell Proliferation; Cells; ChIP-seq; Chromatin; Clinical Research; Cytokine Receptors; DNA; DNA Binding; DNA Binding Domain; DNA Sequence; DNA Tumor Viruses; Data; Dentistry; Dentists; Development; EP300 gene; Environment; Epigenetic Process; Epithelial Cells; Etiology; Florida; Foundations; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic Transcription; Genomic approach; Gingiva; Goals; Growth; Herpesviridae Infections; Histone Acetylation; Human; Human Herpesvirus 8; Immune response; Individual; Infection; Interferons; Kaposi Sarcoma; Knock-out; Luciferases; Lytic; Lytic Phase; Mediating; Mentorship; Metabolism; Methods; Morphogenesis; N Domain; N-terminal; Nuclear; Nuclear Localization Signal; Oncogenic; Oncogenic Viruses; Oral; Oral cavity; Palate Kaposi' s Sarcoma; Pathogenesis; Pathway interactions; Persons; Play; Population; Proteins; Publications; Regulation; Regulator Genes; Reporter; Research; Research Training; Response Elements; Role; Route; Saliva; Salivary; Scientist; Sequence Homology; Sexual Transmission; Signal Pathway; Terminator Codon; Testing; Training; Training Programs; Transcriptional Regulation; Universities; Viral; Viral Genes; Viral Pathogenesis; Virion; Virus; base; career; chromatin immunoprecipitation; chronic infection; college; cytokine; epidemiology study; experimental study; gene induction; histone acetyltransferase; inhibitor; mutant; new therapeutic target; oral biology; oral cavity epithelium; oral infection; pathogen; prevent; promoter; protein complex; protein expression; recruit; small hairpin RNA; transcription factor; transcriptome sequencing; transmission process; tumorigenesis; viral interferon regulatory factor; viral interferon regulatory factor-1; viral transmission; virology

Abstract Infection of humans by the oncogenic Kaposi's sarcoma-associated herpesvirus (KSHV) starts in the oral cavity by replicating in oral epithelial cells. Regulation of lytic KSHV infection of the oral epithelium is still poorly understood, which hampers the development of effective antiviral therapies to block KSHV transmission. One of the unique features of KSHV is that it encodes four different viral interferon regulatory factors (vIRFs), which are homologous with cellular interferon regulatory factors (IRFs). One of them is vIRF1, which is expressed the earliest and at the highest level during KSHV infection and is used by KSHV to control many different aspects of the pathogenesis such as blocking antiviral response or stimulating oncogenesis and the growth of infected cells. While the function of vIRF1 in the inhibition of different immune response signaling pathways have been studied in detail, the transcriptional function of vIRF1 in the regulation of both KSHV and host cellular genes that are important in viral pathogenesis is still poorly understood. vIRF1 is 449 amino acid long protein. It has a putative DNA-binding domain (DBD), which also contains a nuclear localization signal (NLS), and it has a C-terminal IRF interaction domain (IAD). My preliminary experiments revealed that the DNA-binding domain (DBD) of vIRF1 is required for vIRF1-mediated host and viral gene induction, indicating vIRF1 induces both viral and host genes by binding to their promoters. Our research group has also identified the viral and host proteins interacting with vIRF1 by protein complex purification. This revealed the interaction of vIRF1 with histone acetyltransferases (HATs) that often serve as co-factors for various DNA-binding transcription factors in gene induction. I hypothesize that vIRF1 can bind to specific host and viral promoters and uses HATs to induce their corresponding genes to enhance the lytic KSHV infection in oral epithelial cells. I propose two specific aims to test my hypothesis. Aim 1 focuses on identifying the host genes that are directly targeted by vIRF1 in the absence of the other vIRFs in KSHV-infected oral epithelial cells. Aim 2 is to determine the vIRF1 response element in vIRF1 target KSHV promoters that is required for vIRF1-mediated promoter activation. The completion of this project will provide in-depth training in virology, epigenetics and genomics approaches as these methods will be used to elucidate this critical host pathogen interactions. The Comprehensive Training Program in Oral Biology at the University of Florida College of Dentistry will provide me an outstanding environment to prepare me for a career as a dentist scientist by providing personalized mentorship and excellent clinical and research training.

抽象的 致癌性卡波西肉瘤相关疱疹病毒（KSHV）对人类感染开始于口腔 通过在口腔上皮细胞中复制。口服上皮的裂解KSHV感染的调节仍然很差 理解，这阻碍了有效的抗病毒疗法的发展以阻止KSHV传播。之一 KSHV的独特特征是它编码四个不同的病毒干扰素调节因子（VIRF），它们是 与细胞干扰素调节因子（IRF）同源。其中之一是Virf1，它表示 最早，最高水平在KSHV感染期间，KSHV用于控制许多不同方面 发病机理，例如阻断抗病毒反应或刺激肿瘤发生和感染细胞的生长。 虽然已经研究了VIRF1在抑制不同免疫反应信号通路中的功能 详细介绍地，VIRF1在KSHV和宿主细胞基因的调节中的转录功能是 在病毒发病机理中很重要。 VIRF1是449个氨基酸长蛋白。它有一个假定的 DNA结合结构域（DBD），其中还包含核定位信号（NLS），并且具有C端IRF 交互域（IAD）。我的初步实验表明，VIRF1的DNA结合域（DBD）是 VIRF1介导的宿主和病毒基因诱导所必需的，表明VIRF1诱导病毒和宿主基因 通过与其启动子结合。我们的研究小组还确定了病毒蛋白和宿主蛋白与 蛋白质复合物纯化的VIRF1。这揭示了VIRF1与组蛋白乙酰基转移酶的相互作用 （帽子）通常是基因诱导中各种DNA结合转录因子的共同因素。我 假设VIRF1可以与特定的宿主和病毒启动子结合，并使用帽子诱导其 相应的基因，以增强口腔上皮细胞中的裂解KSHV感染。我提出了两个具体目标 检验我的假设。 AIM 1专注于识别在不存在的情况下直接靶向的宿主基因 KSHV感染的口腔上皮细胞中的其他VIRF。 AIM 2是确定在 VIRF1靶标启动子是VIRF1介导的启动子激活所必需的。完成此完成 项目将提供有关病毒学，表观遗传学和基因组学方法的深入培训，因为这些方法将是 用于阐明这种关键的宿主病原体相互作用。口腔生物学的综合培训计划 在佛罗里达大学牙科学院将为我提供一个杰出的环境，为我做好准备 通过提供个性化指导以及出色的临床和研究培训，作为牙医科学家的职业。