Identification and Regulation of RNA Modification by HIV infection and Methamphetamine

项目总结 通过RNA修饰进行转录后基因调控是一个新兴的概念，因为 在信使核糖核酸序列中发现了不同的修饰核苷酸。这样做的总体目标是 建议描绘HIV-1感染和感染的分子和细胞机制 甲基苯丙胺(MA)改变宿主转录和翻译程序导致 免疫系统功能障碍。我们的中心假设是HIV-1感染和MA的使用 改变病毒和宿主表位转录的动态以调节宿主病原体 互动。Rana实验室的最新发现支持这一观点，因为它表明HIV-1 感染对病毒和病毒的甲基化RNA的拓扑结构和功能都有深远的影响 人类的RNA。HIV-1RNA中M6A的丰度由宿主甲基转移酶控制 METTL3和METTL14以及去甲基酶ALKBH5和FTO。然而，基本面和 表位转录调控及其在HIV-1致病机制中作用的关键问题 仍有待解决。为了进行这些研究，我们开发了如下的体外感染模型 以及RNA修饰络合物的生物化学和功能表征。我们的项目 有三个具体目标：目标1：确定病毒和宿主RNA的动态和图谱 艾滋病毒感染和甲基苯丙胺(MA)暴露造成的修饰。目标2：确定如何 RNA修饰影响HIV-1的致病机制。目标3：确定动态和具体 RNA修饰机的功能。这些目标的成功实现将显著 加深我们对表位转录调控基因表达的理解 艾滋病毒感染。此外，这些结果将提供对分子的基本理解。 MA的使用改变了导致免疫功能障碍的机制。

PROJECT SUMMARY Post-transcriptional gene regulation by RNA modification is an emerging concept because a diverse set of modified nucleotides are found in mRNA sequences. The overall objective of this proposal is to delineate the molecular and cellular mechanisms by which HIV-1 infection and methamphetamine (MA) alter host transcriptional and translational programs to cause dysfunction of immune system. Our central hypothesis is that HIV-1 infection and use of MA alter the dynamics of the viral and host epitranscriptomes to regulate host-pathogen interactions. Recent findings from the Rana lab support this notion by showing that HIV-1 infection has profound effects on the topology and function of methylated RNAs of both viral and human RNAs. m6A abundance in HIV-1 RNA is controlled by the host methyltransferases METTL3 and METTL14 and the demethylases ALKBH5 and FTO. However, fundamental and critical questions regarding epitranscriptome regulation and its effects on HIV-1 pathogenesis remain to be addressed. To carry out these studies, we developed in vitro infection models as well as biochemical and functional characterization of RNA modifying complexes. Our project has three specific aims: Aim 1: Define the dynamics and mapping of viral and host RNA modifications by HIV infection and methamphetamine (MA) exposure. Aim 2: Determine how RNA modifications influence HIV-1 pathogenesis. Aim 3: Determine the dynamics and specific functions of RNA modifying machinery. Successful completion of these aims will significantly enhance our understanding of the epitranscriptomic regulation of gene expression caused by HIV infection. In addition, these results will provide fundamental understanding of the molecular mechanisms that are altered by MA use leading to immune dysfunctions.