Epstein-Barr Virus nuclear antigen leader protein in transcription regulation

摘要： 爱泼斯坦-巴尔病毒(EBV)相关疾病仍然是人类健康的巨大负担。作为一种 经口腔传播的病原体，EBV感染可引起传染性单核细胞增多症 各种癌症的病例，包括发生在一个空间内的鼻咽癌 紧邻口腔的是一些B细胞恶性肿瘤，还有约10%的胃癌。 在HIV感染者中，EBV可引起口腔毛状白斑。要了解 EBV促进疾病发展的分子机制， 以EB病毒转化的淋巴母细胞系为模型系统。EB病毒核抗原 前导蛋白(EBNALP)是EB病毒转化幼稚B淋巴细胞所必需的。大部分ITS 已知的功能与EBV转录激活因子EBNA2有关。然而，EBNALP绑定到 许多不依赖于EBNA2的增强子和启动子位点。这些EBNALP的摄动 带有CRISPRi的位点显著降低了这些增强子的相关基因表达。小才是 了解EBNALP如何发挥其EBNA2独立功能。也不知道它是如何 EBNALP与增强子/启动子位置以及它们如何影响转录有关。 因此，我们假设EBNALP利用宿主转录程序获得 获得宿主增强剂/启动子，并通过以下方式促进EBV转化 EBNA2-独立机制。在我的辅导期内，我将向 EBNALP如何与DNA结合的根本问题。我们将使用基于CRISPR的检测来 鉴定EBNALP增强子激活所必需的宿主蛋白。我首先将重点放在 序列特异性转录因子(TF)。染色质免疫沉淀(CHIP) 检测方法将用于检测基因敲除对EBNALP DNA结合的影响。在我的R00期间 阶段，我将独立进行研究，并通过以下方式将我的工作与导师的工作区分开来 研究EBNALP的不同方面。我将把我的研究重点放在增强剂的特性上 EBNALP在增强子上组装蛋白质复合体调控转录 活动。我将重点介绍转录辅助因子、基础转录因子和组蛋白。 修饰酶。了解EBNALP与DNA结合的机制 调节基因转录可能为治疗EBV相关疾病提供有希望的靶点 疾病。

Abstract: Epstein-Barr virus (EBV) associated diseases remain a huge burden in human health. As an orally transmitted pathogen, EBV infection causes infectious mononucleosis and ~200,000 cases of various cancers, including nasopharyngeal carcinoma that occurs in a space immediately adjacent to the oral cavity, some B cell malignancies, and ~10% of gastric cancer. In HIV infected people, EBV causes oral hairy leukoplakia of tongue. To understand the molecular mechanisms through which EBV contributes to disease development, EBV-transformed lymphoblastoid cell lines are used as a model system. EBV nuclear antigen leader protein (EBNALP) is essential for EBV to transform naïve B lymphocytes. Most of its known functions are linked to EBV transcription activator EBNA2. However, EBNALP binds to many enhancer and promoter sites independent of EBNA2. Perturbations of these EBNALP sites with CRISPRi significantly decreased these enhancers’ linked gene expression. Little is known about how EBNALP exerts its EBNA2 independent functions. It is also not known how EBNALP is tethered to the enhancer/promoter sites and how they affect transcription. Therefore, we hypothesize that EBNALP exploits host transcription programs to gain access to host enhancers/promoters, and contributes to EBV transformation through EBNA2-independent mechanisms. During my mentored period, I will address the fundamental question of how EBNALP binds to DNA. We will use CRISPR-based assays to identify host proteins essential for EBNALP enhancer activation. I will first focus on sequence-specific transcription factors (TFs). Chromatin immune precipitation (ChIP) based assays will be used to test the effects of knockout on EBNALP DNA binding. During my R00 phase, I will perform research independently and distinguish my work from my mentor’s by studying different aspects of EBNALP. I will focus my studies on characterizing the enhancer protein complexes assembled by EBNALP onto the enhancers to regulate transcription activity. I will focus on transcription cofactors, basal transcription factors, and histone modifying enzymes. Understanding the mechanisms through which EBNALP binds to DNA and regulates gene transcription may provide promising targets for treating EBV-associated diseases.