Investigating chromatin mechanisms using viral systems

使用病毒系统研究染色质机制

• 批准号: 10649922
• 负责人: Daphne Christina Avgousti
• 金额: $ 5.61万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10649922
• 关键词: 3-Dimensional; Adenovirus Protein; Adenoviruses; Area; Binding; Biological; Biological Process; Biology; Cells; Chromatin; Chromosomes; Development; Disease; Genome; Goals; Herpesviridae; Histones; Infection; Inflammation; Innovative Therapy; Malignant Neoplasms; Molecular Biology; Molecular Conformation; Nuclear; Positioning Attribute; Proteins; RNA Splicing; Regulation; Simplexvirus; System; TP53 gene; Technology; Testing; Variant; Viral; Viral Packaging; Virus; Virus Diseases; cancer therapy; dynamic system; pathogen; virus host interaction

PROJECT SUMMARY All eukaryotic genomes are compacted in a fundamental assembly called chromatin, the aberrant regulation of which is a hallmark of many diseases and cancers. Technologies to study chromatin have advanced significantly in recent years, allowing us to progress from linear sequencing to 3D chromosome conformation studies. However, it is becoming increasingly clear that chromatin states are dynamic and respond to perturbations. Thus a barrier remains to understanding the biological functions of chromatin in a dynamic system, and not merely at the baseline state. Viruses have evolved to take over the cell to produce viral progeny, necessarily hijacking cellular chromatin for viral benefit, thereby providing an ideal dynamic system in which to interrogate chromatin state changes. Moreover, the study of virus-host interactions has been doubly beneficial by leading to advances in virus biology and to discoveries of some of the most important areas of molecular biology, from splicing to p53. Due to the advances in sequencing technology, we are now ideally positioned to take the next step to understanding chromatin in the context of biological changes. In this proposal, we aim to merge cutting-edge chromatin technology with virus infection to reveal fundamental chromatin functions. We use two different nuclear replicating viruses, adenovirus and herpesvirus, as the test cases for virus infection to pinpoint chromatin vulnerabilities exploited by multiple pathogens. We have identified a viral packaging protein from adenovirus, protein VII, that causes global chromatin reorganization and we will systematically dissect the mechanisms by which this reorganization occurs. Our preliminary results suggest that protein VII, together with cellular binding partners, may replace linker histones thus revealing a previously unknown vulnerability of chromatin. We have also uncovered that herpes simplex virus incorporates the histone variant macroH2A1 on its genome during infection. MacroH2A1 has been described as a marker of both silencing and an activating chromatin, therefore, we will use this scenario to untangle the long-standing enigma of macroH2A1 function. Completion of the proposed studies will generate new chromatin targets and facilitate development of innovative therapies for cancer, inflammation, and viral diseases.

项目摘要 所有真核生物的基因组都被压缩在一个叫做染色质的基本集合中， 其调节是许多疾病和癌症的标志。研究染色质的技术 近年来取得了重大进展，使我们能够从线性测序发展到3D测序。 染色体构象研究。然而，越来越清楚的是，染色质状态 动态的并对扰动作出反应。因此，理解生物学功能仍然是一个障碍 在动态系统中的染色质，而不仅仅是在基线状态。病毒已经进化到 在细胞上产生病毒后代，为了病毒的利益，必须劫持细胞染色质， 提供了一个理想的动态系统，其中询问染色质状态的变化。而且 病毒-宿主相互作用的研究具有双重益处，因为它导致了病毒生物学的进步， 发现了分子生物学的一些最重要的领域，从剪接到p53。由于 随着测序技术的进步，我们现在处于理想的位置，可以采取下一步行动， 在生物学变化的背景下理解染色质。在这份提案中，我们的目标是合并 尖端的染色质技术与病毒感染，揭示基本的染色质功能。 我们使用两种不同的核复制病毒，腺病毒和疱疹病毒，作为病毒的测试案例， 感染，以查明染色质漏洞利用多种病原体。我们已经确定了一 来自腺病毒的病毒包装蛋白，即蛋白VII，它引起整体染色质重组， 将系统地剖析这种重组发生的机制。我们的初步结果 这表明蛋白VII与细胞结合伴侣一起可以取代连接体组蛋白， 揭示了染色质的一个未知弱点我们还发现单纯疱疹 病毒在感染过程中将组蛋白变体macroH 2A 1整合到其基因组上。MacroH 2A 1已经 作为沉默和激活染色质的标记，因此，我们将使用这种情况 来解开宏H2 A1功能的长期谜团。完成拟议的研究将 产生新的染色质靶点并促进癌症创新疗法的开发， 炎症和病毒性疾病。