Gene regulatory role of vIRF1 in KSHV infection

vIRF1在KSHV感染中的基因调控作用

基本信息

批准号：
10665555
负责人：
Seung Jin Jang
金额：
$ 4.76万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
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/10665555
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-terminal Nuclear Nuclear Localization Signal Oncogenic Oncogenic Viruses Oral Oral cavity Palate Kaposi&apos s Sarcoma Pathogenesis Pathway interactions Persons Play Population Proteins Publications Regulation Regulator Genes Reporter Repression 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 Virus Replication career chromatin immunoprecipitation chronic infection clinical training cofactor 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

项目摘要

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.

抽象的致癌卡波西肉瘤相关疱疹病毒（KSHV）对人类的感染始于口腔通过在口腔上皮细胞中复制。口腔上皮细胞溶解性 KSHV 感染的调节仍然很差这阻碍了开发有效的抗病毒疗法来阻止 KSHV 传播。之一 KSHV 的独特之处在于它编码四种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，其表达为 KSHV 感染期间最早且处于最高水平，KSHV 利用它来控制 KSHV 的许多不同方面发病机制，例如阻断抗病毒反应或刺激肿瘤发生和受感染细胞的生长。而vIRF1在抑制不同免疫反应信号通路中的功能已被研究详细而言，vIRF1 在 KSHV 和宿主细胞基因调节中的转录功能对于病毒发病机制的重要作用仍知之甚少。 vIRF1 是 449 个氨基酸长的蛋白质。它有一个推定的 DNA 结合域 (DBD)，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需，表明 vIRF1 诱导病毒和宿主基因通过与它们的启动子结合。我们的研究小组还鉴定了与病毒和宿主蛋白相互作用的病毒和宿主蛋白。 vIRF1通过蛋白质复合物纯化。这揭示了 vIRF1 与组蛋白乙酰转移酶的相互作用 (HAT) 通常作为基因诱导中各种 DNA 结合转录因子的辅助因子。我假设 vIRF1 可以与特定宿主和病毒启动子结合，并使用 HAT 来诱导其相应的基因增强口腔上皮细胞中的溶解性KSHV感染。我提出两个具体目标检验我的假设。目标 1 侧重于识别在缺乏 vIRF1 的情况下直接靶向的宿主基因 KSHV 感染的口腔上皮细胞中其他 vIRF 的变化。目标 2 是确定 vIRF1 响应元素 vIRF1 靶向 KSHV 启动子，这是 vIRF1 介导的启动子激活所需的。本次的完成该项目将提供病毒学、表观遗传学和基因组学方法的深入培训，因为这些方法将用于阐明这种关键的宿主病原体相互作用。口腔生物学综合培训计划佛罗里达大学牙科学院将为我提供出色的环境，帮助我为将来的职业生涯做好准备通过提供个性化指导和出色的临床和研究培训，成为一名牙医科学家。