Investigating chromatin mechanisms using viral systems

项目概要 所有真核基因组都压缩在称为染色质的基本组装中，染色质是异常的 其调节是许多疾病和癌症的标志。研究染色质的技术有 近年来取得了显着进步，使我们能够从线性测序进展到 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 动态并对扰动做出响应。因此，理解其生物学功能仍然存在障碍。 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采取 控制细胞产生病毒后代，必然劫持细胞染色质以获取病毒利益，从而 提供一个理想的动态系统来询问染色质状态的变化。此外， 病毒与宿主相互作用的研究带来了双重好处，它促进了病毒生物学的进步， 分子生物学一些最重要领域的发现，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 了解生物变化背景下的染色质。在这个提案中，我们的目标是合并 尖端染色质技术与病毒感染相结合，揭示染色质的基本功能。 我们使用两种不同的核复制病毒，腺病毒和疱疹病毒，作为病毒的测试用例 感染以查明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，蛋白质 VII，导致整体染色质重组，我们 将系统地剖析这种重组发生的机制。我们的初步结果 表明蛋白质 VII 与细胞结合伴侣一起可能取代接头组蛋白，从而 揭示了染色质以前未知的脆弱性。我们还发现，单纯疱疹 病毒在感染过程中将组蛋白变体 MacroH2A1 整合到其基因组中。 MacroH2A1 已 被描述为沉默和激活染色质的标记，因此，我们将使用这种情况 解开长期存在的 MacroH2A1 功能之谜。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、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.