Genome Persistence of KSHV

KSHV 的基因组持久性

基本信息

批准号：
9188807
负责人：
ERLE S. ROBERTSON
金额：
$ 29.88万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-01-01 至 2019-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9188807
关键词：
Adenomatous Polyposis Coli Protein Affinity Chromatography Aneuploidy Animal Model Antigens B-Lymphocytes Biochemical Genetics Biological Assay Body cavities Cell Cycle Cell Proliferation Cells Centromere Centrosome Chromatin Chromosomes Complex Core Protein Cyclin B DNA DNA biosynthesis Data Deletion Mutation Development Disease Elements Endothelial Cells Ensure Episome Etiology Family Fluorescence Resonance Energy Transfer Fluorescent in Situ Hybridization Genes Genome Genomic DNA Goals Graft Rejection HIV Health Herpesviridae Histones Human Human Herpesvirus 8 Imagery In Vitro Kaposi Sarcoma Kinetochores Link Lymphoma Maintenance Malignant Neoplasms Mediating Mitosis Mitotic Mitotic Spindle Apparatus Mitotic spindle Monitor Nuclear Organ Transplantation Phase Phosphorylation Play Pleural effusion disorder Point Mutation Post-Translational Protein Processing Process Proteins RNA Interference Recruitment Activity Regulation Role Series Subfamily lentivirinae Substrate Cycling System Terminal Repeat Sequences Therapeutic immunosuppression Time Trans-Activators Transcriptional Regulation Transplant Recipients Viral Viral Genes Viral Genome Viral Proteins Virus Latency Virus Replication antigen binding aurora B kinase base body cavity cellular targeting cis acting element daughter cell ds-DNA experimental study gammaherpesvirus genetic analysis in vivo insight knock-down latency-associated nuclear antigen latent infection neoplastic cell prevent protein complex protein function public health relevance segregation small hairpin RNA therapeutic development tumor tumorigenesis ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Kaposi's sarcoma associated herpes virus (KSHV/HHV-8) is the second identified gammaherpesvirus associated with at least 3 major proliferative diseases. These include Kaposi's sarcoma and body cavity based lymphomas (BCBLs) or Pleural Effusion Lymphomas (PELs). One of the major latent antigens, latency- associated nuclear antigen (LANA) is constitutively expressed in all latently infected KSHV cells and is involved in the long erm maintenance of the dsDNA KSHV genome as well as transcriptional regulation. Studies have also shown an important role in regulation of viral latency in association with the viral immediate early transactivator Rta. LANA binds to cis-acting elements within the KSHV terminal repeats (TR) and is associated with a number of chromosomal proteins which are important for tethering the viral proteins and segregation of the viral genome to new daughter cells. Furthermore, LANA can induce chromosomal abberations when expressed in human cells in vitro suggesting a role for LANA in induction of oncogenesis in KSHV infected cells. The specific aims of this new proposal is to focus our studies investigating KSHV latent infection, through persistence and mechanism of segregation of the viral genome which is passed from parental cell to progeny cells which are latently infected. There has been no direct studies which comprehensively describes a specific mechanism by which this occurs. We have shown associations of LANA with cellular proteins that are necessary for tethering and segregation of the viral genome in the infected cells. We have also shown an association with the nuclear mitosis apparatus protein NuMA, as well as Bub1 and a number of CenP proteins. Here we will explore the direct connections with LANA and cellular proteins associated with the centromere and kinetochore using state of the art in vivo visualizatin strategies combined with targeted shRNA knockdown of specific cellular targets, to understand the function of these proteins in mediating or controlling the transitional states of the KSHV genomic DNA as it passes from one daughter cell to the next without loss of the genome. We will use direct biochemical, genetics and in vivo FRET visualization analyses to focus on the interaction of LANA with the kinetochore protein Bub1 and the APC proteins known to regulate the stability of this protein complex, and to investigate the mechanism of stability of this protein complex whereby LANA and the KSHV genome can progress through the stages of mitosis without loss of the genome thus allowing segregation of the viral genomes to the new daughter cells.

描述（由申请人提供）：卡波西肉瘤相关疱疹病毒 (KSHV/HHV-8) 是第二种已确定的与至少 3 种主要增殖性疾病相关的伽马疱疹病毒。这些包括卡波西肉瘤和体腔淋巴瘤 (BCBL) 或胸腔积液淋巴瘤 (PEL)。主要潜伏抗原之一，潜伏相关核抗原（LANA）是在所有潜伏感染的 KSHV 细胞中组成型表达，并参与 dsDNA KSHV 基因组的长期维持以及转录调控。研究还表明，在调节与病毒即时相关的病毒潜伏期方面发挥着重要作用。早期反式激活因子 Rta。 LANA 与 KSHV 末端重复序列 (TR) 内的顺式作用元件结合，并与许多染色体蛋白相关，这些蛋白对于束缚病毒蛋白以及将病毒基因组分离到新的子细胞非常重要。此外，LANA 在体外人类细胞中表达时可以诱导染色体畸变，这表明 LANA 在 KSHV 感染细胞中诱导肿瘤发生中发挥作用。这项新提案的具体目标是通过从亲本细胞传递到潜伏感染的子代细胞的病毒基因组的持久性和分离机制，集中研究 KSHV 潜伏感染。目前还没有直接的研究来全面描述这种情况发生的具体机制。我们已经证明了 LANA 与细胞蛋白的关联，这些蛋白对于受感染细胞中病毒基因组的束缚和分离是必需的。我们还显示了与核有丝分裂装置蛋白 NuMA 以及 Bub1 和许多 CenP 蛋白的关联。在这里，我们将使用最先进的体内可视化策略结合特定细胞靶标的靶向 shRNA 敲低，探索与着丝粒和动粒相关的 LANA 和细胞蛋白的直接联系，以了解这些蛋白在介导或控制过渡过程中的功能。 KSHV 基因组 DNA 从一个子细胞传递到下一个子细胞时的状态，且基因组没有丢失。我们将利用直接的生化、遗传学和体内FRET可视化分析来重点研究LANA与动粒蛋白Bub1和已知调节该蛋白复合物稳定性的APC蛋白的相互作用，并研究该蛋白复合物的稳定性机制LANA 和 KSHV 基因组可以通过有丝分裂阶段而不丢失基因组，从而允许病毒基因组分离到新的子细胞。