Mechanisms of HIV-1 RNA Methylation in Regulating Viral Replication

HIV-1 RNA甲基化调节病毒复制的机制

• 批准号: 9315991
• 负责人: Li Wu
• 金额: $ 40.03万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9315991
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Adenosine; Affect; Area; B-Lymphocytes; Binding; Binding Proteins; Binding Sites; Biology; CD4 Positive T Lymphocytes; Cell Line; Cell Proliferation; Cells; Cessation of life; Communicable Diseases; Complex; Data; Development; Frequencies; Gene Expression; Goals; HIV-1; Infection; Maps; Mediating; Messenger RNA; Methyltransferase; Modification; Post-Transcriptional Regulation; Production; Protein Biosynthesis; Protein Family; Proteins; RNA; RNA Binding; RNA Splicing; RNA Stability; RNA methylation; RNA replication; Reader; Regulation; Role; Site; Staging; Testing; Translations; Viral; Viral Reverse Transcription; Virion; Virus; Virus Diseases; Virus Replication; abstracting; base; gag Gene Products; genetic regulatory protein; genomic RNA; inhibitor/antagonist; insight; knock-down; mRNA Expression; mutant; novel; overexpression; promoter; therapeutic development; trafficking; viral RNA

Project Summary/Abstract The goal of this new R01 proposal is to investigate the novel role and mechanisms of N6- methyladenosine (m6A) modification of HIV-1 RNA in viral infection. The internal m6A modification of cellular RNA is a newly emerging mechanism of post-transcriptional control of gene expression, which is coordinately regulated by three groups of host proteins, including adenosine methyltransferases (writers), m6A demethylases (erasers), and m6A-selective- binding proteins (readers). Binding of m6A-modified cellular RNA by the readers, YTH domain family proteins (YTHDF1, 2, and 3), can significantly affect various aspects of RNA functions during translation. However, it is unknown whether HIV-1 RNA contains m6A modification and whether the modification regulates viral replication in CD4+ T-cells. We recently identified multiple regions of m6A modification in HIV-1 genomic RNA (gRNA) bound by the readers (YTHDF1-3 proteins) in HIV-1-infected cells. Our data showed that the expression levels of the readers can significantly modulated HIV-1 postentry infection in target cells. Moreover, knockdown of the m6A writers or an eraser in virus producer cells significantly affected HIV-1 Gag protein synthesis and viral release, suggesting the importance of m6A modification of HIV-1 RNA in viral production. Thus, we hypothesize that m6A modification of HIV-1 RNA regulates viral replication in cells by engaging the m6A readers, writers, and erasers, which can have negative or positive impacts on different stages of the HIV-1 lifecycle. We will test the overall hypothesis in three specific aims mainly using primary CD4+ T-cells. Aim 1. To examine the role of HIV-1 RNA m6A modification in viral infection and the mechanisms of m6A reader-mediated viral inhibition; Aim 2. To investigate the role and mechanisms of the m6A writers in regulating HIV-1 infection; and Aim 3. To investigate the role and mechanisms of the m6A erasers in regulating HIV-1 infection. Overall impact: Accomplishing these proposed studies will reveal the novel role and mechanisms of m6A modification of HIV-1 RNA in regulating viral infection in cells. Studying the m6A modification of HIV-1 RNA and its interactions with host proteins represents a new area of HIV-1 RNA biology, which can facilitate therapeutic development against HIV-1 infection.

项目摘要/摘要 该新R01提案的目的是研究N6-的新作用和机制 HIV-1 RNA在病毒感染中修饰甲基腺苷（M6A）。内部M6A 细胞RNA的修饰是新兴的新兴机制 基因表达，由三组宿主蛋白协调调节，包括 腺苷甲基转移酶（作家），M6a脱甲基酶（橡皮）和M6A选择性 结合蛋白（读取器）。读者yth域对M6a修饰的细胞RNA的结合 家族蛋白（YTHDF1、2和3）可以显着影响RNA功能的各个方面 在翻译过程中。但是，尚不清楚HIV-1 RNA是否包含M6A修饰和 修饰是否调节CD4+ T细胞中的病毒复制。 我们最近确定了HIV-1基因组RNA（GRNA）中M6A修饰的多个区域 受HIV-1感染细胞中的读者（YTHDF1-3蛋白）约束。我们的数据表明 读者的表达水平可以显着调节靶标的HIV-1邮递感染 细胞。此外，在病毒细胞中敲低M6A作者或橡皮擦 受影响的HIV-1 GAG蛋白合成和病毒释放，表明M6A的重要性 HIV-1 RNA在病毒生产中的修饰。因此，我们假设M6A修改 HIV-1 RNA通过吸引M6A读取器，作家和橡皮擦来调节细胞中的病毒复制， 这可能会对HIV-1生命周期的不同阶段产生负面或积极的影响。我们将 在三个特定目的中测试总体假设，主要使用主要的CD4+ T细胞。目标1 检查HIV-1 RNA M6A修饰在病毒感染中的作用和M6A的机制 读者介导的病毒抑制；目标2。研究M6A的作用和机制 调节HIV-1感染的作者；和目标3。调查 M6A橡皮擦调节HIV-1感染。 总体影响：完成这些提出的研究将揭示新作用和 M6A修饰HIV-1 RNA在调节细胞病毒感染中的机制。研究 HIV-1 RNA的M6A修饰及其与宿主蛋白的相互作用代表了一个新区域 HIV-1 RNA生物学，可以促进针对HIV-1感染的治疗发育。