Gene regulatory role of vIRF1 in KSHV infection

vIRF1在KSHV感染中的基因调控作用

• 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
• 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' 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

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.

摘要 致癌性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。口腔上皮细胞的溶解性KSHV感染的调控仍然很差 这阻碍了开发有效的抗病毒疗法来阻断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）， 与细胞干扰素调节因子（IRFs）同源。其中之一是vIRF1，它表达为 在KSHV感染期间最早和最高水平，并被KSHV用于控制许多不同方面的疾病。 发病机制如阻断抗病毒反应或刺激肿瘤发生和感染细胞的生长。 vIRF 1在不同免疫应答信号通路中的抑制作用已被研究 详细地说，vIRF1在KSHV和宿主细胞基因调节中的转录功能， 在病毒发病机制中的重要作用仍然知之甚少。vIRF 1是449个氨基酸长的蛋白质。它有一个假定的 DNA结合结构域（DBD），也含有核定位信号（NLS），C端有一个IRF 相互作用域（IAD）。我的初步实验表明，vIRF1的DNA结合结构域（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。我们的研究小组还确定了病毒和宿主蛋白质相互作用， 通过蛋白复合物纯化vIRF1。这揭示了vIRF1与组蛋白乙酰转移酶的相互作用 在基因诱导中，HAT通常作为各种DNA结合转录因子的辅因子。我 假设vIRF1可以结合特异性宿主和病毒启动子，并利用HAT诱导其 相应的基因来增强口腔上皮细胞中的裂解性KSHV感染。我提出两个具体目标， 验证我的假设目的1侧重于鉴定在缺乏vIRF1的情况下vIRF1直接靶向的宿主基因。 KSHV感染的口腔上皮细胞中其他vIRF的表达。目的2是确定vIRF1反应元件， vIRF1靶向vIRF1介导的启动子激活所需的KSHV启动子。完成这一 该项目将提供病毒学、表观遗传学和基因组学方法方面的深入培训，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。口腔生物学综合训练计划 在佛罗里达大学牙科学院将为我提供一个出色的环境，为我准备一个 通过提供个性化的指导和优秀的临床和研究培训，成为牙医科学家。