Retrovirus Models of Cellular Post-transcriptional Gene Expression

在项目1、2、5以及核心A和B的协作下，项目3阐明了一个新的译文 通过对逆转录病毒基因组的比较研究来调节轴。我们的蛋白质组、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特异性地识别独特的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 依赖大小写的翻译启动。我们的生化分析和全基因组翻译的结果 分析已经确定了生物相关基因的一个子集，需要RHA才能有效地翻译。 许多PCE候选基因是编码复杂的5‘非编码区的原癌基因，这需要RHA/PCE 互动促进高效翻译。我们对RHA在细胞中的基础作用的认识 翻译对照为理解RHA失调是一种肿瘤的观察提供了一个平台 生物标志物。我们的合作研究已经确定了六种不同的逆转录病毒中的PCE活性，包括 HTLV-1；与RHA的相互作用是必要的；这种病毒与宿主的相互作用对于有效的HTLV-1至关重要。 1个译文。我们对细胞结节中PCE活性的额外鉴定提供了概念证明 逆转录病毒采用宿主细胞机制来实现高效的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；这一创新适用于广泛的 基因表达系统阵列。我们的基本见解暗示RHA作为一个综合效应者 基因表达从转录到翻译的连续体以及病毒和细胞基因的协调 表情。最初筹资期间的成果与私营部门方案规划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。这一高度关注的主要焦点 互动继续是为了了解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。具体地说，我们假设RHA调节子是一种可诱导的翻译控制 选定基因的机制，其dvs调节有助于细胞的改变 导致转化和副橡皮病的微环境。我们的三个相互关联的目标是： 目的1.研究RHA基因在细胞中表达和胞质定位的基本特征 周期进展；目的2.检测RHA翻译活性在破骨细胞活性和肿瘤中的作用 模型；目的3.评估与逆转录病毒有关的Jund PCE的基本特征、结构/功能 PCE数据库。长期目标是将RHA转录后调控的知识应用于 开发载体和小分子选择性调节肿瘤相关的RHA反应基因 转化、副肿瘤疾病和逆转录病毒感染。

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.