Epitranscriptomic regulation of HBV gene expression

HBV基因表达的表观转录组调控

基本信息

批准号：
10092086
负责人：
ALEEM SIDDIQUI
金额：
$ 39.46万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10092086
关键词：
3&apos Untranslated Regions 5&apos Untranslated Regions Adenosine Affinity Antiviral Agents Applications Grants Binding Bioinformatics Biological Assay Biological Process Capsid Cell Nucleus Cells Chronic Chronic Hepatitis Chronic Hepatitis B Circular DNA Code Core Protein DNA DNA biosynthesis Data Development Enzymes Gene Expression Genetic Transcription Genome Genomic DNA Health Hepatitis B Virus Homeostasis Human Immune response Immunoprecipitation Infection Inflammation Length Life Cycle Stages Liver Liver diseases Messenger RNA Methods Methylation Methyltransferase Modification Monitor Mutate Mutation Nucleosome Core Particle Nucleotides Obesity Pathogenesis Patients Play Poly(A)+ RNA Positioning Attribute Primary carcinoma of the liver cells Process Proteins RNA RNA Viruses RNA methylation Regulation Reporting Research Resolution Reverse Transcription Risk Factors Role Scheme Site Stress Structure System Techniques Terminator Codon Tissues Transcript Translations Vaccines Viral Viral Genome Viral Proteins Virus Virus Diseases Virus Replication Withdrawal base design epitranscriptomics liver biopsy therapeutically effective transcriptome viral DNA viral RNA

项目摘要

Project Summary/Abstract Human hepatitis B virus (HBV causes chronic hepatitis. Chronic hepatitis B is a major risk factor in the progression to the onset of hepatocellular carcinoma (HCC). In spite of an effective vaccine and currently used antivirals, there are estimated 350 million infected carriers worldwide. Antivirals do not eliminate HBV DNA in the nucleus. A better understanding of the role of host and viral factors contributing to chronic hepatitis is needed. Epitranscriptomic regulation of HBV gene expression provides another layer of regulatory schemes of gene expression. In this study, we propose to characterize the posttranscriptional modification of viral RNAs at the N6 position of the adenosine base termed m6A. Our extensive preliminary studies described in the grant application show that HBV RNAs are methylated as m6A. Using the strategy of silencing of host methylases and demethylase enzymes involved in epitranscriptomic homeostasis, we present evidence that HBV RNAs are m6A modified. There was clearly an effect on translation of viral transcripts in the absence of enzyme that catalyze m6A modification. Our results with MeRIP immunoprecipitation followed by Tru-seq analysis identified a potential range of sequences enriched in a peak of m6A residues. We mutated about 13 `A' residues in this region that spans from nucleotides 1700 to 2000 within the HBV genome. These were examined for translation of HBV proteins and reverse transcription function. Results were striking, in that, the translation was negatively regulated but reverse transcription was positively regulated by m6A modification. Based on these exciting data, we propose to continue characterization of m6A modification of HBV RNAs and identify the functional relevance of this modification in the HBV infection. This study opens a new avenue of HBV research to investigate epitranscriptomic regulation of the unique HBV life cycle and possible contribution to the progression to chronicity associated with infection.

项目概要/摘要人类乙型肝炎病毒（HBV导致慢性肝炎。慢性乙型肝炎是慢性乙型肝炎的主要危险因素进展至肝细胞癌（HCC）的发生。尽管有有效的疫苗并且目前正在使用据估计，全球有 3.5 亿感染者。抗病毒药物不能消除乙肝病毒 DNA 核。更好地了解宿主和病毒因素对慢性肝炎的作用需要。 HBV 基因表达的表观转录组调控提供了另一层调控方案基因表达。在这项研究中，我们建议表征病毒 RNA 的转录后修饰腺苷碱基的 N6 位置称为 m6A。我们在拨款中描述了广泛的初步研究应用表明 HBV RNA 被甲基化为 m6A。采用沉默宿主甲基化酶的策略和去甲基化酶参与表观转录组稳态，我们提供证据表明 HBV RNA m6A 已修改。在缺乏酶的情况下，病毒转录物的翻译明显受到影响催化m6A修饰。我们通过 MeRIP 免疫沉淀和 Tru-seq 分析确定了结果富含 m6A 残基峰的潜在序列范围。我们在此突变了大约 13 个“A”残基该区域横跨 HBV 基因组内的核苷酸 1700 至 2000。这些都经过了检查 HBV 蛋白的翻译和逆转录功能。结果是惊人的，因为翻译是 m6A 修饰负向调节逆转录，但逆转录受到正向调节。基于这些令人兴奋的数据，我们建议继续表征 HBV RNA 的 m6A 修饰并确定这种修饰在 HBV 感染中的功能相关性。这项研究开辟了乙型肝炎研究的新途径研究独特的 HBV 生命周期的表观转录组调控及其对进展为与感染相关的慢性病。