Investigating chromatin mechanisms using viral systems

项目总结 所有真核生物的基因组都被压缩在一个称为染色质的基本组件中，这是一种异常 对它的调控是许多疾病和癌症的标志。研究染色质的技术有 近年来取得了显著的进步，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，染色质状态是 动态的，并对扰动做出反应。因此，理解生物功能仍然是一个障碍。 染色质在动态系统中的变化，而不仅仅是在基线状态。病毒已经进化到可以 在细胞上产生病毒后代，必须劫持细胞染色质以获得病毒益处，从而 提供了一种询问染色质状态变化的理想动态系统。此外， 病毒-宿主相互作用的研究带来了病毒生物学的进步和 分子生物学的一些最重要领域的发现，从剪接到P53。由于 测序技术的进步，我们现在处于理想的地位，可以采取下一步 在生物变化的背景下理解染色质。在这项提案中，我们的目标是合并 先进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。 感染以确定被多种病原体利用的染色质漏洞。我们已经确定了一个 来自腺病毒的病毒包装蛋白，蛋白VII，引起全球染色质重组和WE 将系统地剖析这种重组发生的机制。我们的初步结果 提示蛋白VII与细胞结合伙伴一起，可能因此取代连接子组蛋白 揭示了染色质以前未知的弱点。我们还发现了单纯疱疹病毒 病毒在感染过程中将组蛋白变异体宏H_2A1整合到其基因组上。宏观H_2A1已经被 被描述为沉默和激活染色质的标记，因此，我们将使用这个场景 以解开长期存在的宏H_2A1功能之谜。建议的研究完成后， 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。

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.