Viral Long Noncoding RNA Functions in Epstein-Barr Virus Infection

病毒长非编码 RNA 在 Epstein-Barr 病毒感染中的功能

• 批准号: 9017925
• 负责人: JEFFERY T SAMPLE
• 金额: $ 37.83万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9017925
• 关键词: Acquired Immunodeficiency Syndrome; Adoption; B-Lymphocytes; Binding Proteins; CCCTC-binding factor; Cells; Cellular Immunity; Chromatin; Chromatin Loop; Complex; Control Locus; DNA replication origin; Data; Development; Disease; Distal; Epstein-Barr Virus Infections; Epstein-Barr Virus latency; Equilibrium; Event; Gene Duplication; Gene Expression; Genes; Genetic Transcription; Genome; Herpesviridae; Human; Human Herpesvirus 4; Immune; Individual; Infection; Infectious Mononucleosis; Introns; Knowledge; Life; Location; Lymphoma; Lymphoproliferative Disorders; Lytic Phase; Maintenance; Malignant Neoplasms; Mediating; Medical; Memory B-Lymphocyte; Messenger RNA; MicroRNAs; Molecular Conformation; Nucleotides; Oncogenic; Patients; Pattern; Play; Process; Property; Proteins; RNA; RNA Splicing; Recombinants; Regulation; Research; Rest; Risk Factors; Role; Small Nucleolar RNA; Testing; Transcript; Untranslated RNA; Viral; Viral Genes; Viral Genome; Viral Proteins; Virus; Virus Latency; Virus Replication; cofactor; defined contribution; immunosuppressed; latency-associated protein; latent infection; paralogous gene; pathogen; persistent EBV infection; programs; promoter; protein expression; public health relevance; trafficking; viral RNA

DESCRIPTION (provided by applicant): Epstein-Barr virus (EBV) is an extremely successful pathogen, being able to establish a lifelong latent infection within B lymphocytes of its human host, with overt disease in healthy individuals restricted to a self- limiting mononucleosis in ~40% of individuals when infection is delayed until the second decade of life. However, a breakdown in cellular immunity, particularly as a consequence of AIDS, remains a significant risk factor for development of EBV-associated lymphoma and lymphoproliferative disease, underscoring the highly evolved equilibrium that exists between EBV and its host. Establishment of this equilibrium, and EBV's oncogenic potential are dependent on a highly coordinated and controlled expression of the EBV latency- associated genes, which encode not only proteins but at least four classes of noncoding RNAs (ncRNAs). The long-term objective of the research proposed in this application is to understand the contributions made to EBV infection and pathogenic potential by two EBV RNAs belonging to the class of long ncRNAs (lncRNAs), i.e., RNAs longer than 200 nucleotides with little or no functional protein-encoding capability. These lncRNAs are encoded by the paralogous EBV genes BHLF1 and LF3, long known to be expressed at high level during the virus replication or lytic cycle, but recently identified as beig actively expressed during latent infection as well. Our preliminary results indicate that, upon deletion of the BHLF1 locus from the EBV genome, the virus is unable to sustain full expression of the EBV latency-associated proteins, converting to a more restricted program of latency-gene expression associated with long-term EBV persistence within its cellular reservoir, the memory B cell. Thus, we hypothesize that the BHLF1 lncRNA, and its paralog from LF3, play active but distinct roles in the regulation of EBV latent infection and persistence. We propose four specific aims: Under Aim 1, we will identify proteins that interact with the BHLF1 lncRNA, and the role that these specific RNA:protein associations play in EBV latent infection. In Aim 2 we will elucidate the expression patterns of the BHLF1 lncRNAs during the different EBV latency programs (defined by specific patterns of viral gene expression), and whether they traffic to specific subcellular domains with known functionality, possibly revealing function(s) of the EBV lncRNA. In Aim 3 we will elucidate whether the BHLF1 gene locus itself, in concert with its lncRNA, acts as a central determinant of EBV latency program by regulating viral genome configuration and other mechanisms. Finally, under Aim 4 we will begin to define the roles that the LF3 locus and lncRNA contribute to EBV latency, taking a similar tack as we have proposed for BHLF1.

描述(申请人提供)：爱泼斯坦-巴尔病毒(EBV)是一种非常成功的病原体，能够在其宿主的B淋巴细胞内建立终身潜伏感染，在健康个体中的显性疾病限制在大约40%的个体中，当感染被推迟到生命的第二个十年时。然而，细胞免疫功能的崩溃，特别是由于艾滋病，仍然是EBV相关淋巴瘤和淋巴增殖性疾病发展的一个重要风险因素，突显了EBV和其宿主之间存在的高度进化的平衡。这种平衡的建立和EBV的致癌潜力依赖于EBV潜伏期相关基因的高度协调和受控表达，这些基因不仅编码蛋白质，而且至少编码四类非编码RNA(NcRNAs)。本申请中提出的研究的长期目标是了解属于长ncRNAs(LncRNAs)的两个EBV RNAs对EBV感染和致病潜力的贡献，即长于200个核苷酸的RNAs，其功能蛋白编码能力很低或没有。这些LncRNAs由相似的EBV基因BHLF1和LF3编码，长期以来人们知道它们在病毒复制或裂解周期中高水平表达，但最近发现在潜伏感染期间也有BEIG活跃表达。我们的初步结果表明，当BHLF1基因从EBV基因组中删除后，病毒不能维持EBV潜伏期相关蛋白的完全表达，而是转换为与其细胞储存库-记忆B细胞内的EBV长期持续相关的潜伏期基因表达的更受限制的程序。因此，我们假设BHLF1 lncRNA及其与LF3的平行序列在EBV潜伏感染和持久性的调节中发挥着积极而独特的作用。我们提出了四个特定的目标：在目标1下，我们将识别与BHLF1 lncRNA相互作用的蛋白质，以及这些特定的RNA：蛋白质关联在EBV潜伏感染中所起的作用。在目标2中，我们将阐明BHLF1 lncRNA在不同EBV潜伏期程序中的表达模式(由病毒基因表达的特定模式定义)，以及它们是否运输到具有已知功能的特定亚细胞域，可能揭示EBV lncRNA的功能(S)。在目标3中，我们将阐明BHLF1基因位点本身是否与其lncRNA一起，通过调节病毒基因组配置和其他机制作为EBV潜伏期程序的中心决定因素。最后，在目标4下，我们将开始定义LF3基因座和lncRNA对EBV潜伏期的作用，采取与我们为BHLF1提出的类似策略。