Effects of HIV Rev on Host Cell Gene Expression

HIV Rev 对宿主细胞基因表达的影响

• 批准号: 10546602
• 负责人: MARIE-LOUISE HAMMARSKJOLD
• 金额: $ 28.26万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10546602
• 关键词: Affect; Algorithms; Alternative Splicing; Binding; Bioinformatics; CD4 Positive T Lymphocytes; Cell physiology; Cells; Coupled; Cytoplasm; Data Analyses; Development; Electrophoretic Mobility Shift Assay; Elements; Endogenous Retroviruses; Gene Expression; Genes; Genetic Transcription; HIV; HIV Infections; Human; Immune response; In Vitro; Infection; Introns; Lead; Libraries; Mediating; Messenger RNA; Methods; Natural Immunity; Paper; Peripheral; Play; Post-Transcriptional Regulation; Process; Protein Isoforms; RNA; RNA Sequences; RNA Splicing; Response Elements; Retroviral Vector; Retroviridae; Role; Structure; System; T-Lymphocyte; Testing; Tissue-Specific Gene Expression; Transcript; Translating; Translations; Untranslated RNA; Viral; Virus Replication; bioinformatics tool; cell growth regulation; cellular transduction; cis acting element; experimental study; in vivo; mRNA Expression; novel; novel therapeutics; response; rev Protein; shape analysis; transcriptome; transcriptome sequencing; vector; viral RNA

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.

众所周知，所有逆转录病毒都进化出了克服通常的调控机制的过程。 细胞控制或限制RNA核质输出直至剪接完成的机制。一 每种逆转录病毒转录组的标志是具有至少一个保留内含子的病毒mRNA到达 细胞质在HIV感染细胞的情况下，出现了许多不同的带有一个或两个保留内含子的病毒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，促进了这一过程。本提案将探讨以下假设， 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， 元件存在于真正的细胞基因和人类内源性逆转录病毒，以克服 通常控制和限制具有保留内含子的RNA同种型输出的机制。新型RNA 其表达已被Rev失调的同种型可能起到改变感染者的环境的作用。 细胞影响先天免疫和/或促进病毒复制，或者直接地，或者通过翻译成新的 蛋白质异构体。 该提案有两个具体目标： 在目标#1中，我们将鉴定与Rev功能性相互作用的细胞RNA。 通过使用一种新的矢量捕获系统。这将允许分离细胞序列， 如RRE（cRRE），并确定有可能被Rev调节的基因。在目标#2中，我们将执行 用表达Rev或达特和Rev两者的逆转录病毒载体转导的SupT 1细胞上的RNASeq，并鉴定 具有在细胞质中表达的新型RNA同种型的基因。 在完成这项R21研究时，我们预计已经确定了HIV Rev是否可以诱导 通过与cRRE的相互作用，在转录后水平上的新RNA种类的细胞质表达。 进一步的实验，超出了这个两年的建议的范围，将分析这种新的Rev介导的 细胞RNA的转录后调节影响HIV复制和细胞功能。