Viral Noncoding RNAs and Cell Transformation

病毒非编码 RNA 和细胞转化

基本信息

批准号：
10553131
负责人：
JOAN A. STEITZ
金额：
$ 67.28万
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/10553131
关键词：
2019-nCoV 3&apos Untranslated Regions Adopted B-Lymphocytes Binding Biogenesis Biological Callithrix Cancer Patient Cebidae Cell Nucleus Cell physiology Cells Collaborations 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 Polyadenylation Population Primates Process Production Protein Biosynthesis Protein Engineering Proteins Proteome 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 cell growth regulation cell transformation combat crosslink design differential expression expectation 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.

项目概要/摘要携带三种致癌疱疹病毒的淋巴细胞中非编码 (nc)RNA 的作用正在被研究研究表明，EB 病毒 (EBV) 会感染并转化人类 B 细胞；传染性单核细胞增多症，与多种人类癌症有关。在新大陆猴体内产生致命性淋巴瘤和白血病，并在培养物中转化人类 T 淋巴细胞。卡波西肉瘤相关疱疹病毒 (KSHV) 折磨免疫功能低下的个体，并在体内持续存在近年来，我们将研究重点放在了其结构和功能上。两个 EBV 编码的 EBER、七个 HVS 编码的 HSUR 和六个 HVS microRNA，以及 KSHV PAN RNA。这些病毒 ncRNA 含量丰富，在相关病毒之间保守，并与宿主蛋白结合。我们的功能研究揭示了 microRNA 生物发生和衰变的新机制，揭示病毒 ncRNP 对于病毒 DNA 复制等多种核过程至关重要 (EBER2) 或 mRNA 输出到细胞质 (PAN)，确定了三螺旋在 RNA 稳定中的作用，以及最引人注目的是我们对病毒 ncRNA 的研究已经为病毒进化提供了重要的见解。发现了细胞的存在并开始阐明新的细胞机制，例如细胞的调节 microRNA 群体以及 PolyA 尾和 3'UTR 如何协作稳定细胞 mRNA。拟议的目标将利用这些进展进一步研究潜在的分子机制。将扩展我们最初发现的靶向 microRNA 衰减 (TDMD) 来识别蛋白质和有助于细胞 microRNA 降解的其他 RNA 信号，并研究推测源自 SARS-CoV-2 基因组的小 RNA 在调节宿主免疫反应中具有潜在作用我们将确定其多聚腺苷酸尾巴以及内部序列的诊断/治疗意义。有助于 PAN RNA 实现晚期裂解 mRNA 核输出的能力，从而产生病毒颗粒蛋白我们将搜索细胞转录组数据库中是否存在 RNA。有助于polyA-3'UTR相互作用（大概还有RNA稳定）的序列/结构基序，如我们最近通过高分辨率 X 射线分析发现了新发现的三链体形成元素 (ENE)。将分析冠状病毒 RNA 的稳定活性以及对病毒蛋白的可能贡献 EBV-病毒转录物之间具有广泛的相互作用。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。