Genetic and Biochemical Studies of the KSHV LANA Gene

KSHV LANA 基因的遗传和生化研究

• 批准号: 6898492
• 负责人: Kenneth M Kaye
• 金额: $ 39.21万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6898492
• 关键词: DNA replication; Kaposi' s sarcoma; affinity chromatography; cell line; flow cytometry; gene deletion mutation; gene therapy; genetic regulatory element; histones; human herpesvirus 8; immunofluorescence technique; intermolecular interaction; latent virus infection; nuclear proteins; nucleic acid repetitive sequence; nucleosomes; plasmids; point mutation; protein structure function; transfection /expression vector; virus DNA; virus antigen; virus genetics; yeast two hybrid system

DESCRIPTION (provided by applicant): Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) is etiologically linked with KS, primary effusion lymphomas (PELs) and multicentric Castleman's disease. These diseases occur in AIDS and other patients and current therapies are limited. KSHV latently infects the vast majority of tumor cells and viral DMA persists as a multiple copy, extrachromosomal, circular episome. To persist in proliferating cells, episomes must replicate and efficiently segregate to daughter nuclei. KSHV latency associated nuclear antigen (LANA) binds terminal repeat (TR) DNA to mediate KSHV episome persistence. LANA mediates TR DMA replication and tethers episomes to mitotic chromosomes for efficient segregation to progeny nuclei. Significant gaps remain in our understanding of LANA mediated episome maintenance. LANA chromosome association is essential for episome tethering yet the mechanisms underlying chromosome attachment are ill-defined. LANA is a large, multifunctional protein. Although LANA shares sequence homology and episome maintenance function with LANA homologs of other gamma-2 herpesviruses, it is much larger than the other LANA homologs. Definition of the LANA domains required for episome maintenance is central to understanding KSHV episome persistence. The precise cis-acting TR sequence requirements for LANA mediated DNA replication and episome maintanence are unknown. Since TRs account for -20% of the KSHV genome, an understanding of the necessary cis-acting episome persistence sequence is fundamental to a better understanding of KSHV biology. This work will investigate critical aspects of LANA's episome maintenance function. The mechanisms by which the LANA N- and C-termini independently associate with chromosomes will be elucidated. The LANA domains required for episome persistence will be identified. The minimal cis-acting TR sequence requirements for LANA mediated episome persistence will be defined. Since episome persistence is critical for latent infection and most tumor cells are latently infected, this work may lead to new strategies for prevention and treatment of KSHV associated malignancies. Further, definition of the LANA and cis-acting element requirements for episome maintenance will enhance construction of LANA based gene therapy vectors.

描述(申请人提供)：卡波西肉瘤(KS)相关疱疹病毒(KSHV)在病因学上与KS、原发渗出性淋巴瘤(PEL)和多中心Castleman病有关。这些疾病发生在艾滋病和其他患者身上，目前的治疗方法有限。KSHV潜伏感染绝大多数肿瘤细胞，而病毒DMA以多拷贝、染色体外、环状集落的形式持续存在。为了维持增殖的细胞，上皮体必须复制并有效地分离到子核。KSHV潜伏期相关核抗原(LANA)与末端重复序列(TRDNA)结合，介导KSHV Episome持久性。LANA介导TRDMA复制，并将表观染色体系在有丝分裂染色体上，以便有效地分离到后代核。 在我们对LANA介导的Episome维持的理解上仍然存在显著的差距。Lana染色体联合对于Episome系留是必不可少的，然而染色体附着的机制还不清楚。LANA是一种大的多功能蛋白质。虽然LANA与其他Gamma-2疱疹病毒的LANA同源物具有序列同源性和Episome维护功能，但它比其他LANA同源物要大得多。定义Episome维持所需的LANA结构域是理解KSHV Episome持久性的核心。LANA介导的DNA复制和Episome维持所需的确切顺式作用TR序列尚不清楚。由于TRs占KSHV基因组的-20%，了解必要的顺式作用Episome持续序列是更好地理解KSHV生物学的基础。 这项工作将调查LANA的Episome维护功能的关键方面。LANA的N-末端和C-末端与染色体独立结合的机制将被阐明。将确定Episome持久性所需的LANA域。将定义LANA介导的Episome持久性的最小顺式作用的TR序列要求。由于EB病毒的持久性对潜伏感染至关重要，而且大多数肿瘤细胞都是潜伏感染的，这项工作可能会导致预防和治疗KSHV相关恶性肿瘤的新策略。此外，定义LANA和Episome维持所需的顺式作用元件将加强基于LANA的基因治疗载体的构建。