Effects of HIV Rev on Host Cell Gene Expression

众所周知，所有逆转录病毒都有进化过程来克服通常的调控 在剪接完成之前，控制或限制核质输出核糖核酸的细胞机制。一个 每个逆转录病毒转录组的特点是至少有一个保留内含子的病毒mRNA到达 细胞质。在HIV感染的细胞中，出现了许多带有一个或两个保留内含子的不同病毒RNA 在细胞质中，它是REV蛋白与顺式作用元件REV反应元件的结合 (RRE)，存在于所有这些RNA中，促进这一过程。这项提议将探索这样的假设， 除了促进带有保留内含子的病毒RNA的输出和表达外，REV还与RNA相互作用 存在于真正的细胞基因和人类内源性逆转录病毒中的元件，以克服 正常情况下控制和限制内含子保留的RNA异构体出口的机制。新型核糖核酸 其表达被REV失调的异构体可能会改变感染者的环境 细胞影响先天免疫和/或促进病毒复制，无论是直接的，还是通过翻译成小说 蛋白质异构体。 该提案有两个具体目标： 在目标1中，我们将识别在功能上与Rev.相互作用的细胞RNA。这将实现 通过使用一种新型的媒介诱捕系统。这将允许分离起作用的细胞序列 像RRES(CRRES)一样，并确定有可能受Rev.调控的基因。在目标2中，我们将执行 表达REV或同时表达TAT和REV的逆转录病毒载体转导的SupT1细胞上的RNAseq 具有新的RNA异构体的基因在细胞质中表达。 在R21完成时，我们预计已经确定HIV Rev是否可以诱导 通过与CRRES的相互作用在转录后水平上表达新的RNA物种。 在这两年计划的范围之外，进一步的实验将分析这部小说是如何由REV介导的 细胞RNA的转录后调控影响HIV复制和细胞功能。

It is well established that all retroviruses have evolved processes to overcome the usual regulatory mechanisms of the cell that control or restrict the nucleocytoplasmic export of RNA until splicing is complete. A hallmark of every retroviral transcriptome is that viral mRNA with at least one retained intron reaches the cytoplasm. In the case of HIV infected cells, many different viral RNAs with one or two retained introns appear in the cytoplasm, and it is the binding of the Rev protein to a cis-acting element, the Rev Response Element (RRE), present in all of these RNAs, that facilitates this process. This proposal will explore the hypothesis that, in addition to facilitating the export and expression of viral RNAs with retained introns, Rev interacts with RNA elements present in bona fide cellular genes and human endogenous retroviruses, to overcome the mechanisms that normally control and restrict the export of RNA isoforms with retained introns. Novel RNA isoforms, whose expression has been dysregulated by Rev, may function to change the milieu of the infected cell to affect innate immunity and/or promote viral replication, either directly, or by being translated into a novel protein isoforms. The proposal has 2 specific aims: In Aim #1 we will identify cellular RNAs that functionally interact with Rev. This will be accomplished through the use of a novel vector-trap system. This will allow the isolation of cellular sequences that function like RREs (cRREs) and identify genes with potential to be regulated by Rev. In Aim #2 we will perform RNASeq on SupT1 cells transduced with retroviral vectors that express Rev or both Tat and Rev and identify genes that have novel RNA isoforms expressed in the cytoplasm. At the completion of this R21, we expect to have determined whether HIV Rev can induce the cytoplasmic expression of novel RNA species at the post-transcriptional level through interaction with cRREs. Further experiments, beyond the scope of this two year proposal, will analyze how this novel Rev-mediated post-transcriptional regulation of cellular RNAs influences HIV replication and cellular functions.