N6-methyladenosine and the biology of HIV

N6-甲基腺苷和 HIV 生物学

基本信息

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
项目类别：
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

项目摘要

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（应用程序提供）：最近的技术进步表明，在人mRNA和长的非编码RNA中，M6A甲基化丰富，可以指导广泛的细胞过程。 HIV基因组包括许多推定的M6A修饰位点，我们的初步实验数据表明，M6A修饰在HIV基因组中介绍。拟议的研究的总体目标是指出HIV基因组RNA中M6Amixification的精确部位，并确定这些修饰对HIV生物学的功能影响。在AIM 1中，我们将采用特定位点的裂解和放射性标记，然后进行连接辅助提取和薄层色谱法（Scarlet），以量化并精确定位单个核丁基分辨率的M6A修饰。在AIM 2中，我们将研究M6A RNA修饰对HIV感染和病毒产生的影响。此外，我们将测试受感染的单核细胞衍生的树突状细胞的HIV中不同M6A对HIV的免疫原性的影响。提出的实验将表征HIV RNA的RNA修饰的新型模式。 HIV M6A修饰的表征及其在HIV生命周期中的作用不仅会扩大我们对HIV的了解，还会扩大M6A修饰的其他病毒的了解。