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装中，染色质的异常调节 这是许多疾病和癌症的标志。研究染色质的技术已经有了长足的进步 近年来，使我们能够从线性测序发展到三维染色体构象研究。 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， 要理解染色质在动态系统中的生物学功能仍然存在障碍，而不仅仅是在 基线状态。病毒已经进化到接管细胞以产生病毒后代，这必然会劫持 细胞染色质对病毒有益，从而提供了一种理想的动态系统，其中询问染色质 状态发生变化。此外，对病毒-宿主相互作用的研究也因取得进展而双管齐下 病毒生物学和分子生物学中一些最重要的领域的发现，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。在这项提议中，我们的目标是将尖端技术 染色质技术与病毒感染，揭示染色质的基本功能。我们用两个 不同的核复制病毒，腺病毒和疱疹病毒，作为病毒感染的检测案例 染色质漏洞被多种病原体利用。我们已经从一种病毒包装蛋白中鉴定出 腺病毒，蛋白VII，引起全球染色质重组，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VII与 细胞结合伙伴，可以取代连接子组蛋白，从而揭示了以前未知的脆弱性 染色质。我们还发现，单纯疱疹病毒结合了组蛋白变异体宏H_2A1。 它的基因组在感染过程中。MacroH_2A1被描述为沉默和激活的标志 染色质，因此，我们将利用这个场景来解开长期存在的大分子H2A1功能之谜。 拟议研究的完成将产生新的染色质目标，并促进创新的 癌症、炎症和病毒性疾病的治疗。

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.