Viral Noncoding RNAs and Cell Transformation

病毒非编码 RNA 和细胞转化

• 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
• 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' 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

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在携带三种致癌疱疹病毒的淋巴细胞中的作用是 调查过了。爱泼斯坦-巴尔病毒(EBV)感染并转化人类B细胞；它是引起 传染性单核细胞增多症，与几种人类癌症有关。人类疱疹病毒(HVS)诱导 新大陆猴子的致命性淋巴瘤和白血病，并在培养中转化人类T淋巴细胞。 卡波西肉瘤相关疱疹病毒(KSHV)困扰着免疫功能低下的人，并持续在 潜伏的形式，直到溶解激活。近年来，我们将重点放在了结构和功能上 两个EBV编码的EBER，七个人类免疫系统编码的HSURs和六个人类免疫系统的microRNA，以及KSHV PAN RNA。这些病毒的ncRNAs都是丰富的，在相关病毒之间保守，并结合宿主蛋白与 表格ncRNPs。我们的功能研究揭示了microRNA生物发生和衰退的新机制， 揭示了病毒的核糖核酸核糖核酸在多种核过程中都是必不可少的(EBER2)。 或信使核糖核酸输出到细胞质(PAN)，确定三螺旋在RNA稳定中的作用，以及 为病毒进化提供了重要的见解。最引人注目的是，我们对病毒ncRNA的研究 发现并开始阐明新的细胞机制的存在，如细胞的调节 MicroRNA群体以及Polya Tail和3‘UTR如何合作稳定细胞mRNAs。 提出的AIMS将利用这些进展来进一步研究潜在的分子机制。我们 将扩展我们最初发现的靶向microRNA衰变(TDMD)以识别蛋白质和 其他有助于细胞microRNA降解的RNA信号，以及研究A 来自SARS-CoV-2基因组的推测的小RNA调节宿主免疫反应，具有潜在的 诊断/治疗的含义。我们将确定它的Polya尾部以及内部序列 有助于PAN RNA实现晚期裂解mRNAs的核输出，导致病毒粒子蛋白 生产和释放病毒粒子。我们将在细胞转录组数据库中搜索RNA的存在 有助于Polya-3‘UTR相互作用(以及可能的RNA稳定)的序列/结构基序，如 最近我们的高分辨率X射线分析揭示了。新发现的三链形成元素(烯) 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中病毒转录本之间的广泛相互作用 通过补骨脂素交联发现的感染细胞将得到验证和进一步分析。