Viral Noncoding RNAs and Cell Transformation

病毒非编码 RNA 和细胞转化

基本信息

批准号：
10364830
负责人：
JOAN A. STEITZ
金额：
$ 64.35万
依托单位：
YALE UNIVERSITY
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：
2022-01-20 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10364830
关键词：
2019-nCoV 3&apos Untranslated Regions Adopted B-Lymphocytes Binding Biogenesis Biological Callithrix Cancer Patient Cebidae Cell Nucleus Cell physiology Cells Coronavirus Cytoplasm DNA biosynthesis Databases Diagnostic Elements Epstein-Barr virus encoded RNA 2 Evolution Gene Expression Genome Goals Herpesviridae Human Human Herpesvirus 4 Human Herpesvirus 8 Immune response Immune signaling Immunocompromised Host In Vitro Individual Infection Infectious Agent Infectious Mononucleosis Investigation Lymphoid Cell Lymphoma Lytic Lytic Phase Malignant Neoplasms Mass Spectrum Analysis Measurement Messenger RNA MicroRNAs Molecular Monkeys Mutagenesis Nuclear Nuclear Export Oncogenic Poly A Population Primates Process Production Protein Biosynthesis Protein Engineering Proteins Proteome Psoralens RNA RNA Databases RNA Sequences RNA Stability RNA-Binding Proteins Resolution Ribonucleoproteins Roentgen Rays Role SARS-CoV-2 genome SARS-CoV-2 infection Saimiriine Herpesvirus 2 Shapes Signal Transduction Small RNA Structure System T-Lymphocyte Tail Testing Therapeutic Transcript Untranslated RNA Viral Viral Proteins Virion Virus cell growth regulation cell transformation combat crosslink design differential expression expectation gammaherpesvirus gene function insight leukemia/lymphoma mRNA Export messenger ribonucleoprotein mutant novel sarcoma transcriptome transforming virus triple helix viral DNA viral rescue

项目摘要

Project Summary/Abstract The roles of noncoding (nc)RNAs in lymphoid cells harboring each of three oncogenic herpesviruses are being investigated. Epstein-Barr virus (EBV) infects and transforms human B cells; it is the causative agent of infectious mononucleosis and is associated with several human cancers. Herpesvirus saimiri (HVS) induces fatal lymphomas and leukemias in New World monkeys and transforms human T lymphocytes in culture. Kaposi's sarcoma-associated herpesvirus (KSHV) afflicts immunocompromised individuals and persists in a latent form until lytic activation. In recent years, we have focused our efforts on the structure and functions of the two EBV-encoded EBERs, the seven HVS-encoded HSURs and six HVS microRNAs, as well as the KSHV PAN RNA. These viral ncRNAs are all abundant, conserved between related viruses and bind host proteins to form ncRNPs. Our functional studies have uncovered novel mechanisms of microRNA biogenesis and decay, revealed that viral ncRNPs can be essential for nuclear processes as diverse as viral DNA replication (EBER2) or mRNA export to the cytoplasm (PAN), identified the role of triple helices in RNA stabilization, and contributed important insights into viral evolution. Most compelling is that our studies of viral ncRNAs have uncovered the existence of and begun to elucidate novel cellular mechanisms such as the regulation of cellular microRNA populations and how the polyA tail and 3′UTR may collaborate to stabilize cellular mRNAs. Proposed aims will exploit these advances to further investigate the underlying molecular mechanisms. We shall extend our original discovery of target-directed microRNA decay (TDMD) to identify proteins and additional RNA signals contributing to cellular microRNA degradation, as well as investigate the role of a putative small RNA derived from the SARS-CoV-2 genome in regulating host immune responses, with potential diagnostic/therapeutic implications. We shall establish how its polyA tail as well as internal sequences contribute to PAN RNA's ability to enable the nuclear export of late lytic mRNAs, leading to virion protein production and virion release. We shall search cellular transcriptome databases for the presence of RNA sequence/structure motifs contributing to polyA-3′UTR interactions (and presumably RNA stabilization), as recently revealed by our high-resolution X-ray analyses. Newly discovered triplex-forming elements (ENEs) in coronavirus RNAs will be analyzed for their stabilization activity and possible contributions to viral protein synthesis, with potential therapeutic applications. Extensive interactions between viral transcripts in EBV- infected cells discovered by psoralen crosslinking will be validated and further analyzed.

项目摘要/摘要非编码（NC）RNA在具有三个致癌疱疹病毒中每一个的淋巴样细胞中的作用正在调查。爱泼斯坦 - 巴尔病毒（EBV）感染并转化人类B细胞；它是传染性单核细胞增多症，并与几种人类癌症有关。疱疹病毒Saimiri（HVS）影响新世界猴子的致命淋巴瘤和白血病，并改变培养物中的人类T淋巴细胞。卡波西（Kaposi）与肉瘤相关的疱疹病毒（KSHV）折磨了免疫功能低下的个体，并坚持潜在形式直到裂解激活。近年来，我们将精力集中在这两个EBV编码的Ebers，七个HVS编码的HSUS和六个HVS microRNA以及KSHV PAN RNA。这些病毒ncRNA都是丰富的，在相关病毒和宿主蛋白之间构成与形成ncrnps。我们的功能研究发现了microRNA生物发生和衰减的新型机制，表明病毒NCRNP对于像病毒DNA复制一样潜水的核过程至关重要（EBER2）或导出到细胞质（PAN）的mRNA，确定了三螺旋在RNA稳定中的作用，并且对病毒进化有了重要的见解。最引人注目的是我们对病毒NCRNA的研究具有发现并开始阐明新型细胞机制，例如细胞的调节 MicroRNA种群以及Polya尾巴和3'UTR如何合作以稳定细胞mRNA。拟议的目标将探索这些进步，以进一步研究潜在的分子机制。我们应扩展我们最初发现目标指导的microRNA衰减（TDMD），以识别蛋白质和导致细胞microRNA降解的其他RNA信号，并研究了A的作用在控制宿主免疫反应中，源自SARS-COV-2基因组的假定小RNA，具有潜力诊断/治疗意义。我们将确定其polya尾巴和内部序列如何有助于PAN RNA实现晚期裂解mRNA的核出口的能力，从而导致病毒体蛋白生产和病毒率释放。我们将搜索蜂窝转录组数据库以存在RNA 序列/结构基序有助于polya-3'utr相互作用（大概是RNA稳定），如最近，我们的高分辨率X射线分析揭示了。新发现的三成型元素（ENE）将分析冠状病毒RNA的稳定活性和对病毒蛋白的可能贡献合成，具有潜在的治疗应用。 EBV中的病毒转录本之间的广泛相互作用由牛皮岩交联发现的感染细胞将得到验证并进一步分析。