N6-methyladenosine and the biology of HIV

N6-甲基腺苷和 HIV 生物学

• 批准号: 9151914
• 负责人: TAO PAN
• 金额: $ 26.19万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-25 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9151914
• 关键词: Adenosine; Affect; Apoptosis; Biology; Cell Nucleus; Cell physiology; Cells; Chemicals; Chickens; Cytoplasm; Data; Dendritic Cells; Dependency; Development; Figs - dietary; Future; Genome; Goals; HIV; HIV Genome; Human; Immune; Immune Tolerance; Immune response; Infection; Knock-out; Knowledge; Label; Life Cycle Stages; Ligation; Mammals; Maps; Mediating; Messenger RNA; Methods; Methylation; Methyltransferase; Modification; Nuclear; Nucleotides; Pathway interactions; Pattern; Phase; Pluripotent Stem Cells; Positioning Attribute; Production; Proteins; RNA; RNA Splicing; RNA Viruses; RNA methylation; Radioactive; Reader; Recruitment Activity; Research; Resolution; Retroviridae; Role; Rous sarcoma virus; Site; Somatic Cell; T-Lymphocyte; Testing; Thin Layer Chromatography; Transcript; Translations; Untranslated RNA; Viral; Virus; Virus Replication; base; cell type; embryonic stem cell; fitness; genomic RNA; high reward; high risk; immunogenic; immunogenicity; improved; knock-down; monocyte; novel; novel strategies; novel therapeutic intervention; public health relevance; research study; stoichiometry; transcriptome

 DESCRIPTION (provided by applicant): Recent technological advances showed that m6A methylation is abundant in human mRNA and long non- coding RNA and can guide a wide range of cellular processes. The HIV genome comprises numerous putative m6A modification sites and our preliminary experimental data show that m6A modifications are present in the HIV genome. The overall goal of the proposed research is to pinpoint the precise sites of m6Amodifications within the HIV genomic RNA and to determine the functional impact of these modifications on HIV biology. In Aim 1 we will apply Site-specific Cleavage And Radioactive-labeling followed by Ligation-assisted Extraction and Thin-layer chromatography (SCARLET), to quantify and precisely locate m6A modifications at a single nucleotide resolution. In Aim 2 we will study the effects of m6A RNA modifications on HIV infectivity and viral production. In addition, we will test the effect of varying m6A in HIV on the immunogenicity of HIV by infecting monocyte derived dendritic cells. The proposed experiments will characterize a novel mode of RNA modification of HIV RNA. The characterization of HIV m6A modifications and its elucidation of its role in the HIV life cycle would not only expand our knowledge of HIV, but also for other viruses that are m6A modified.

 描述（由申请人提供）：最近的技术进步表明，m6A甲基化在人类mRNA和长非编码RNA中丰富，并且可以指导广泛的细胞过程。 HIV 基因组包含许多假定的 m6A 修饰位点，我们的初步实验数据表明 HIV 基因组中存在 m6A 修饰。拟议研究的总体目标是查明 HIV 基因组 RNA 内 m6A 修饰的精确位点，并确定这些修饰对 HIV 生物学的功能影响。在目标 1 中，我们将应用位点特异性切割和放射性标记，然后应用连接辅助提取和薄层色谱 (SCARLET)，以单核苷酸分辨率定量和精确定位 m6A 修饰。在目标 2 中，我们将研究 m6A RNA 修饰对 HIV 感染性和病毒产生的影响。此外，我们将通过感染单核细胞衍生的树突状细胞来测试HIV中不同的m6A对HIV免疫原性的影响。拟议的实验将表征 HIV RNA 的 RNA 修饰的新模式。 HIV m6A 修饰的表征及其在 HIV 生命周期中的作用的阐明不仅将扩展我们对 HIV 的了解，而且还将扩展我们对 m6A 修饰的其他病毒的了解。