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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要这项新 R01 提案的目标是研究 N6- 的新作用和机制病毒感染中 HIV-1 RNA 的甲基腺苷 (m6A) 修饰。内部m6A 细胞RNA修饰是一种新兴的转录后控制机制基因表达，由三组宿主蛋白协调调节，包括腺苷甲基转移酶（写入酶）、m6A 去甲基酶（擦除酶）和 m6A-选择性- 结合蛋白（阅读器）。阅读器 YTH 结构域与 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 功能的各个方面翻译过程中。然而，目前尚不清楚 HIV-1 RNA 是否含有 m6A 修饰以及该修饰是否调节 CD4+ T 细胞中的病毒复制。我们最近在 HIV-1 基因组 RNA (gRNA) 中发现了多个 m6A 修饰区域被 HIV-1 感染细胞中的阅读器（YTHDF1-3 蛋白）结合。我们的数据表明 Reader的表达水平可以显着调节目标中的HIV-1进入后感染细胞。此外，病毒生产细胞中 m6A 写入器或擦除器的敲低显着影响 HIV-1 Gag 蛋白合成和病毒释放，表明 m6A 的重要性病毒生产中 HIV-1 RNA 的修饰。因此，我们假设 m6A 修饰 HIV-1 RNA 通过参与 m6A 读取器、写入器和擦除器来调节细胞中的病毒复制，这可能对 HIV-1 生命周期的不同阶段产生负面或正面影响。我们将主要使用原代 CD4+ T 细胞在三个特定目标中检验总体假设。目标 1. 至研究HIV-1 RNA m6A修饰在病毒感染中的作用以及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 感染的作家；目标 3. 研究 m6A 橡皮擦调节 HIV-1 感染。总体影响：完成这些拟议的研究将揭示新的作用和 HIV-1 RNA m6A 修饰调节细胞病毒感染的机制。正在学习 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领域 HIV-1 RNA 生物学，可以促进针对 HIV-1 感染的治疗方法的开发。