Investigating chromatin mechanisms using viral systems

项目总结 (如家长奖助金所述) 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进步 重要的是，近年来，使我们能够从线性测序发展到3D染色体构象 学习。然而，越来越清楚的是，染色质状态是动态的，并对 微扰。因此，要理解染色质在动态变化中的生物学功能仍然存在障碍。 系统，而不仅仅是处于基线状态。病毒已经进化到接管细胞以产生病毒 子代，必须劫持细胞染色质以获得病毒益处，从而在 以询问染色质状态的变化。此外，对病毒-宿主相互作用的研究也加倍 有益于导致病毒生物学的进步和发现一些最重要的领域 分子生物学，从剪接到P53。由于测序技术的进步，我们现在最理想的是 准备采取下一步，在生物变化的背景下理解染色质。在这 建议，我们的目标是将尖端的染色质技术与病毒感染相结合，以揭示 基本染色质功能。我们使用两种不同的核复制病毒，腺病毒和 疱疹病毒作为病毒感染的测试案例，以查明多个利用的染色质漏洞 病原体。我们已经从腺病毒中鉴定出一种病毒包装蛋白，即蛋白VII，它能引起全球 染色质重组，我们将系统地剖析这种重组的机制 发生。我们的初步结果表明，蛋白VII与细胞结合伙伴一起，可能会取代 连接子组蛋白因此揭示了染色质的一种以前未知的脆弱性。我们还发现， 单纯疱疹病毒在感染过程中在其基因组上整合了组蛋白变异体宏H_2A1。宏观H_2A1 被描述为沉默和激活染色质的标志，因此，我们将使用这个 解开长期存在的宏氢氧化铝功能之谜的情景。建议的研究完成后， 产生新的染色质靶点并促进癌症、炎症、 和病毒性疾病。

PROJECT SUMMARY (as stated in parent grant) 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.