E2-Cellular Complexes in HPV Chromatin Transcription

HPV 染色质转录中的 E2 细胞复合物

基本信息

批准号：
8627560
负责人：
CHENG-MING CHIANG
金额：
$ 38.4万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8627560
关键词：
Address Anogenital venereal warts Apoptosis Automobile Driving Binding Binding Proteins Binding Sites Biological Biological Assay Bovine Papillomavirus Bromodomain Bromodomain Containing Protein 2 CREB-binding protein Cell Cycle Arrest Cell Differentiation process Cells Cervix carcinoma Chromatin Collaborations Complex Cutaneous DNA Sequence DNA biosynthesis Development Disease EP300 gene Enzymes Epithelial Cells Exhibits FOS gene Family member Florida Funding G9a histone methyltransferase Gelatinase B Gene Activation Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Transcription Genome HPV-High Risk Head and Neck Cancer Hela Cells Human Papilloma Virus Vaccine Human Papillomavirus Human papilloma virus infection Human papillomavirus 11 Human papillomavirus 16 ITGB4 gene Immunization In Vitro Individual Integrins JUN gene Lead Link Low risk HPV Malignant neoplasm of cervix uteri Malignant neoplasm of penis Maps Mediating Molecular Nuclear Extract Nucleic Acid Regulatory Sequences Nucleosomes Oncogene Proteins Oncogenic Pathogenesis Pattern Phase Phosphorylation Population Protein Family Proteins Recombinants Recruitment Activity Relative (related person)Repression Research Research Personnel Risk Role Site Skin Skin Cancer Synthesis Chemistry System Tertiary Protein Structure Testing Therapeutic Therapeutic Agents Topoisomerase II Transcription Factor AP-1 Transcription Repressor/Corepressor Transcriptional Activation Transcriptional Regulation Vaccines Viral Viral Genes Viral Genome Virus base boys cancer cell carcinogenesis casein kinase II chromatin immunoprecipitation drug development genome wide association study genome-wide girls high risk histone acetyltransferase human CREBBP protein human disease in vivo inhibitor/antagonist innovation malignant mouth neoplasm member non-oncogenic promoter prophylactic public health relevance reconstitution segregation senescence therapeutic vaccine viral DNA virus host interaction

项目摘要

DESCRIPTION (provided by applicant): Human papillomaviruses (HPVs) are etiological agents for many human diseases, including genital warts, skin warts, cervical cancer, and certain types of head-and-neck and skin cancers. Although two prophylactic vaccines against HPV infection are presently available and recommended for immunization of young girls/boys, therapeutic vaccines that are most needed for the majority of HPV-infected populations are still lacking. Since early protein 2 (E2) encoded by HPVs is crucial for viral DNA replication, HPV genome segregation, control of viral gene expression, and induction of cellular apoptosis and senescence, it has become a promising target for the development of anti-viral therapeutics. In the last funding period, we have identified bromodomain- containing protein 4 (Brd4) as a central cellular factor mediating diverse functions of HPV-encoded E2 protein, particularly via formation of an E2-Brd4 silencing complex able to block activator protein-1 (AP-1)-triggered gene activation from latent HPV chromatin that exhibits no transcriptional activity. In this renewal application, we propose three aims to dissect the mechanisms of HPV chromatin-dependent transcriptional regulation: 1. To define the mechanism of AP-1-dependent HPV chromatin transcription 2. To define the repression mechanism of E2-Brd4 in HPV chromatin transcription 3. To define HPV-specific versus global cellular gene transcription regulated by AP-1, Brd4, and E2 We will first apply an innovative HPV chromatin transcription system that we have developed to define the role of cellular chromatin-modifying enzymes, including p300 and CBP histone acetyltransferases (HATs), in AP-1-dependent activation of HPV chromatin transcription. We will then dissect the steps of E2-Brd4 silencing complex in suppressing HPV chromatin transcription and further elucidate the functional role of two newly identified Brd4-interacting proteins, i.e., G9a histone methyltransferase (HMT) and topoisomerase II¿-binding protein 1 (TopBP1), in E2-regulated HPV transcription. To distinguish HPV-specific and global host cell gene transcription, we will also apply genome-wide approaches, in collaboration with Dr. Bing Ren at UC-San Diego, to analyze the transcriptional profiles and binding patterns of these HPV transcriptional regulators and chromatin modifiers, and with the help from Dr. Thomas Kodadek at Florida Scripps, we will utilize synthetic chemistry to develop small compounds blocking Brd4 action in HPV transcription. Our central hypothesis here is that conformational changes in Brd4 provide a molecular switch for Brd4 recruitment of distinct cellular factors to regulate HPV-specific and cellular gene transcription. The combination of mechanistic studies and genome-wide analyses will significantly advance our understanding of viral and cellular factors modulating HPV transcription and facilitate the development of therapeutic agents blocking HPV-induced human diseases.

描述（由申请人提供）：人乳头瘤病毒（HPV）是许多人类疾病的病原体，包括生殖器疣、皮肤疣、宫颈癌以及某些类型的头颈癌和皮肤癌，尽管有两种针对 HPV 感染的预防性疫苗。目前已提供并推荐用于年轻女孩/男孩的免疫接种，但由于早期蛋白 2，大多数 HPV 感染人群最需要的治疗性疫苗仍然缺乏。 HPV编码的E2对于病毒DNA复制、HPV基因组分离、病毒基因表达的控制以及细胞凋亡和衰老的诱导至关重要，它已成为开发抗病毒疗法的有希望的靶标。在此期间，我们发现含溴结构域蛋白 4 (Brd4) 是介导 HPV 编码的 E2 蛋白多种功能的中心细胞因子，特别是通过形成 E2-Brd4 沉默复合物，能够阻断激活蛋白-1 (AP-1) 触发不具有转录活性的潜伏 HPV 染色质的基因激活在本次更新申请中，我们提出了三个目标来剖析 HPV 染色质依赖性转录调控机制： 1. 明确该机制。 AP-1 依赖性 HPV 染色质转录的影响 2. 定义 E2-Brd4 在 HPV 染色质转录中的抑制机制 3. 定义 HPV 特异性与全局细胞AP-1、Brd4 和 E2 调控的基因转录我们将首先应用我们开发的创新 HPV 染色质转录系统来定义细胞染色质修饰酶（包括 p300 和 CBP 组蛋白乙酰转移酶 (HAT)）在 AP- 中的作用HPV 染色质转录的 1 依赖性激活然后我们将剖析 E2-Brd4 沉默复合物抑制 HPV 染色质的步骤。转录并进一步阐明两个新鉴定的 Brd4 相互作用蛋白的功能作用，即 G9a 组蛋白甲基转移酶 (HMT) 和拓扑异构酶 II¿结合蛋白 1 (TopBP1)，在 E2 调节的 HPV 转录中为了区分 HPV 特异性转录和全局宿主细胞基因转录，我们还将与加州大学圣地亚哥分校的 Bing Ren 博士合作，应用全基因组方法来区分 HPV 特异性转录和全局宿主细胞基因转录。分析这些 HPV 转录调节因子和染色质修饰剂的转录谱和结合模式，并在佛罗里达州斯克里普斯的 Thomas Kodadek 博士的帮助下，我们将利用合成化学来开发阻断 Brd4 作用的小化合物HPV 转录。我们的中心假设是，Brd4 的构象变化为 Brd4 招募不同的细胞因子提供了分子开关，以调节 HPV 特异性和细胞基因转录。机制研究和全基因组分析的结合将显着增进我们对 HPV 转录的理解。病毒和细胞因子调节 HPV 转录并促进开发阻断 HPV 诱发的人类疾病的治疗药物。