Adenovirus manipulation of cellular chromatin to overcome host responses

腺病毒操纵细胞染色质以克服宿主反应

• 批准号: 10238103
• 负责人: Matthew D. Weitzman
• 金额: $ 54.24万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-24 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10238103
• 关键词: Address; Adenovirus Infections; Adenovirus Protein; Adenoviruses; Antiviral Agents; Antiviral Response; Appearance; Architecture; Binding; Biochemical; Biological Models; Cell Nucleus; Cell physiology; Cells; ChIP-seq; Chromatin; Chromatin Structure; Core Protein; DNA; DNA Binding; DNA Damage; DNA Viruses; Data; Disease; Effectiveness; Enterobacteria phage P1 Cre recombinase; Evolution; Excision; Gene Delivery; Gene Expression; Genetic Transcription; Genome; Genomics; Goals; HMGB1 gene; Health; Histones; Host Defense; Human; Immune response; Immunoblotting; Infection; Inflammatory; Innate Immune Response; Integration Host Factors; Interferon Activation; Interferons; Knowledge; Lead; Major Core Protein; Malignant Neoplasms; Modification; Molecular; Mutation; Nature; Nucleic Acids; Nucleosomes; Oncolytic; Outcome; Physical condensation; Plaque Assay; Play; Production; Proteins; Proteomics; Public Health; Quantitative Reverse Transcriptase PCR; Resources; Role; Signal Transduction; Stimulus; System; Testing; Therapeutic; Vaccination; Viral; Viral Core Proteins; Viral Genes; Viral Genome; Viral Packaging; Viral Proteins; Virion; Virus; Virus Diseases; design; experimental study; genome-wide; human DNA; improved; novel; particle; pathogen; protein expression; protein function; recruit; response; sensor; tool; vector; viral DNA

PROJECT SUMMARY The need to evade antiviral immune responses has driven evolution of viral strategies to manipulate host cell chromatin, to control gene expression and subvert cellular defenses. Adenoviruses are ubiquitous viruses that have provided both important model systems for understanding fundamental cellular processes, as well as vectors for therapeutic purposes. Infections by Adenovirus and derived vectors elicit strong innate immune responses, which impact both the course of disease and their effectiveness as vectors for gene delivery and vaccination. Therefore understanding viral proteins that subvert host responses is important for treating infections and improving therapeutic vector applications. Here we focus on Adenovirus protein VII, the major core protein that was previously thought to function exclusively for packaging viral genomes. We discovered that protein VII has unexpected roles on both viral and host genomes during Adenovirus infection. The long- term goal of this project is to decipher how this histone-like viral protein overcomes innate host responses and promotes production of infectious viral progeny. We recently showed that protein VII binds DNA and nucleosomes, possesses post-transcriptional modifications (PTMs) analogous to histones, and accumulates in chromatin during infection. We demonstrated that protein VII can alter the composition of host proteins associated with viral genomes in the nuclei of infected cells, and also sequesters host factors in cellular chromatin. Our central hypothesis is that Adenovirus protein VII mimics histones as part of an insidious strategy that manipulates both viral and cellular chromatin to subvert innate host responses. This novel hypothesis has been formulated on the basis of extensive preliminary data produced in our lab with new tools we have generated to study how protein VII is necessary and sufficient to counter cellular immune responses. The two integrated Specific Aims are designed to test our hypothesis by studying functions of protein VII on viral and cellular genomes. Aim 1 will define functions of protein VII on the viral genome during infection. Biochemical and molecular experiments will address how protein VII exploits host machinery to promote production of infectious progeny. Aim 2 will examine the impacts of protein VII on cellular chromatin and genome architecture. Genomic and cellular approaches will determine how association of protein VII with cellular proteins and the chromatinized genome serves to counteract antiviral host responses. We have assembled an interdisciplinary collaborative team to determine this core viral histone-like protein subverts cellular innate immune responses in a project that has broader implications for understanding how viruses elicit changes in chromatin to overcome host defenses.

项目摘要 逃避抗病毒免疫反应的需求导致病毒策略的发展以操纵宿主细胞 染色质，以控制基因表达并颠覆细胞防御。腺病毒是无处不在的病毒 已经提供了重要的模型系统，以了解基本的蜂窝过程以及 用于治疗目的的向量。腺病毒和衍生媒介的感染引起了强烈的先天免疫 反应，这会影响疾病的进程及其作为基因递送向量的有效性和 疫苗接种。因此，了解颠覆宿主反应的病毒蛋白对于治疗很重要 感染和改善治疗载体应用。在这里，我们专注于腺病毒蛋白VII，主要 核心蛋白以前被认为仅用于包装病毒基因组。我们发现了 该蛋白质VII在腺病毒感染期间在病毒和宿主基因组中具有意外作用。长期 该项目的术语目标是破译这种类似组蛋白的病毒蛋白如何克服先天宿主的反应和 促进传染性病毒后代的产生。我们最近表明蛋白质VII结合DNA和 核小体具有类似于组蛋白的转录后修饰（PTM），并积累 感染过程中的染色质。我们证明蛋白质VII可以改变宿主蛋白的组成 与感染细胞核中的病毒基因组相关，并且还隔离了细胞中的宿主因子 染色质。我们的中心假设是腺病毒蛋白VII模仿组蛋白是阴险的一部分 操纵病毒和细胞染色质以颠覆先天宿主反应的策略。这本小说 假设已根据我们实验室中的广泛初步数据提出了假设 我们已经生成了蛋白质VII如何必要且足以应对细胞免疫反应。 这两个集成的特定目的旨在通过研究蛋白质VII的功能来检验我们的假设 病毒和细胞基因组。 AIM 1将在感染期间定义蛋白质VII在病毒基因组上的功能。 生化和分子实验将解决蛋白质VII如何利用宿主机械促进 产生传染性后代。 AIM 2将检查蛋白质VII对细胞染色质和 基因组建筑。基因组和细胞方法将决定蛋白质VII的缔合如何与 细胞蛋白和染色化基因组可抵消抗病毒宿主反应。我们有 组装了一个跨学科的合作团队，以确定这种核心病毒组蛋白样蛋白 在一个项目中，细胞先天免疫反应对了解病毒的引起有更广泛的影响 染色质的变化以克服宿主防御。