Retrovirus Models of Cellular Post-transcriptional Gene Expression

细胞转录后基因表达的逆转录病毒模型

• 批准号: 7876676
• 负责人: Kathleen A. Boris-Lawrie
• 金额: $ 20.34万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-21 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7876676
• 关键词: 5' Untranslated Regions; Adopted; Biochemical; Biological; Biological Markers; Biological Models; Cell Cycle Progression; Cell model; Cells; Cellular biology; Code; Collaborations; Comparative Study; Complex; Databases; Disease; Disease Progression; Elements; Evaluation; Exhibits; Funding; Gene Expression; Gene Expression Regulation; Gene Transfer; Genes; Genetic; Genetic Transcription; Genome; Genus Alpharetrovirus; Goals; Growth; Human T-lymphotropic virus 1; Indium; Internal Ribosome Entry Site; JUN gene; Knowledge; Laboratories; Link; Lymphocyte; Lymphocyte Activation; Malignant Neoplasms; Mediating; Messenger RNA; Modeling; Molecular Cloning; Neoplastic Cell Transformation; Osteoclasts; Outcome; Output; Phase; Post-Transcriptional Regulation; Protein Biosynthesis; Proteins; Proteome; Proteomics; Proto-Oncogenes; Publications; RNA; RNA helicase A; Reagent; Regulation; Regulon; Research; Retroviral Vector; Retroviridae; Retroviridae Infections; Role; Serum; Small Interfering RNA; Structure; System; Taxes; Testing; Translation Initiation; Translational Regulation; Translations; Untranslated Regions; Viral; Virus; cell transformation; deprivation; genome-wide; innovation; insight; lymphocyte proliferation; novel; research study; small molecule; transgene expression; tumor; tumor growth; vector; virus host interaction

In collaboration with Projects 1, 2, 5 and Cores A and B, Project 3 has elucidated a new translation regulatory axis by comparative studies of retroviral genomes. Our results of proteomic, genetic and biochemical analysis have identified a novel role for RNA helicase A (RHA) in both retroviral and cellular genes. We have shown that RHA specifically recognizes a unique 5' terminal post-transcriptional control element (PCE) and neutralizes structural features of the 5' untranslated region (UTR) to facilitate efficient cap-dependent translation initiation. Our results of biochemical analyses and genome-wide translation profiling have identified a subset of biologically-related genes that require RHA for their efficient translation. Many of these PCE candidates are proto-oncogenes encoding a complex 5' UTR, which require RHA/PCE interaction to promote efficient translation. Our identification of the fundamental role of RHA in cellular translational control provides a platform to understand the observation that RHA dysregulation is a tumor biomarker. Our collaborative studies have identified PCE activity in six divergent retroviruses, including HTLV-1; that interaction with RHA is necessary; and this virus-host interaction is essential for efficient HTLV- 1 translation. Our additional identification of PCE activity in cellular junD provided proof-of-concept that retroviruses have adopted a host cell mechanism to achieve efficient RNA expression. We have applied the PCE/host interaction to stimulate protein output in retroviral vectors; this innovation is applicable to a wide array of gene expression systems. Our fundamental insights implicate RHA as an integrative effector in the continuum of gene expression from transcription to translation and in coordinating viral and cellular gene expression. The outcomes of the initial funding period are inextricably linked to the common PPG goal to understand virus-host interactions and mechanisms of gene regulation. A primary focus of this highly interactive Continuation is to understand the scope and regulation of the RHA/PCE translational control axis in retroviral and host genes. Specifically, we postulate the RHA regulon is an inducible translational control mechanism of selected genes, whose dvsregulation contributes to alterations of the cellular microenvironment leading to transformation and paraneoolastic disease. Our three interrelated Aims are: Aim 1. To characterize essential features of RHA gene expression and cytoplasmic localization during cell cycle progression; Aim 2. To examine role of RHA translational activity in osteoclast activity and Tax tumor model; Aim 3. To assess the essential features structure/function of junD PCE in relation to the retrovirus PCE database. Long-term objectives are application of knowledge of the RHA post-transcriptional regulon to develop vectors and small molecules to selectively modulate RHA responsive genes involved in neoplastic transformation, paraneoplastic disease and retrovirus infection.

项目 3 与项目 1、2、5 以及核心 A 和 B 合作，阐明了新的翻译 通过逆转录病毒基因组的比较研究来确定调控轴。我们的蛋白质组学、遗传学和 生化分析确定了 RNA 解旋酶 A (RHA) 在逆转录病毒和细胞中的新作用 基因。我们已经证明 RHA 特异性识别独特的 5' 末端转录后控制 元件（PCE）并中和5'非翻译区（UTR）的结构特征，以促进高效 依赖于帽的翻译起始。我们的生化分析和全基因组翻译结果 分析已经确定了需要 RHA 才能有效翻译的生物学相关基因的子集。 许多这些候选 PCE 是编码复杂 5' UTR 的原癌基因，需要 RHA/PCE 互动促进高效翻译。我们对 RHA 在细胞中的基本作用的鉴定 翻译控制提供了一个平台来理解 RHA 失调是一种肿瘤的观察结果 生物标志物。我们的合作研究已经确定了六种不同逆转录病毒的 PCE 活性，包括 HTLV-1；与 RHA 的互动是必要的；这种病毒与宿主的相互作用对于高效 HTLV 至关重要 1 翻译。我们对细胞 junD 中 PCE 活性的额外鉴定提供了概念验证： 逆转录病毒采用宿主细胞机制来实现有效的RNA表达。我们已经应用了 PCE/宿主相互作用刺激逆转录病毒载体中的蛋白质输出；这项创新适用于广泛 基因表达系统阵列。我们的基本见解表明 RHA 作为一个综合效应器 从转录到翻译以及协调病毒和细胞基因的基因表达连续体 表达。初始资助期的结果与 PPG 的共同目标密不可分 了解病毒与宿主的相互作用和基因调控机制。这个高度的主要焦点 互动延续是了解RHA/PCE平移控制轴的范围和调节 在逆转录病毒和宿主基因中。具体来说，我们假设 RHA 调节子是一种诱导型翻译控制 选定基因的机制，其dvsregulation有助于改变细胞 导致转化和副细胞疾病的微环境。我们的三个相互关联的目标是： 目的 1. 表征细胞发育过程中 RHA 基因表达和细胞质定位的基本特征 循环进展；目标 2. 检查 RHA 翻译活性在破骨细胞活性和 Tax 肿瘤中的作用 模型;目标 3. 评估 junD PCE 与逆转录病毒相关的基本特征结构/功能 PCE数据库。长期目标是将 RHA 转录后调节子的知识应用到 开发载体和小分子来选择性调节与肿瘤相关的 RHA 反应基因 转化、副肿瘤性疾病和逆转录病毒感染。