Genetic and Biochemical Studies of the KSHV LANA Gene

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，病毒DNA以多拷贝、染色体外、环状集落的形式存在。有几条证据支持这样一种模型，即潜伏期相关核抗原(LANA)通过首先介导KSHV DNA的复制，然后将Eisome与染色体捆绑在一起，从而有效地分离到后代核来维持Eisome。因此，阻断LANA的Episome持久性功能的有效干预有望预防或消除KSHV感染，并为KSHV相关的恶性肿瘤提供有效的治疗。理解LANA介导的KSHV Episome持久性的机制对于设计筛选能够抑制这些过程的小分子至关重要。此外，了解LANA DNA结合域与同源DNA的复合结构将使合理的药物设计能够干扰LANA的功能。这项工作将研究LANA的Episome维持功能的核心机制。将确定介导LANA与染色体关联的蛋白质。LANA介导的KSHV DNA复制的机制将被研究。由于LANA的C末端与KSHV DNA特异结合，因此将开始研究与同源DNA结合的这个结构域的结构。这些目标中的每一个都研究了对LANA Episome维护至关重要的区域。由于EB病毒的持久性是潜伏感染的核心，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或治疗KSHV相关恶性肿瘤的新策略。

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 DNA persists as a multiple copy, extrachromosomal, circular episome. Several lines of evidence support a model in which the latency associated nuclear antigen (LANA) maintains episomes by first mediating replication of KSHV DNA and then tethering episomes to chromosomes for efficient segregation to progeny nuclei. Therefore, effective interventions which interrupt LANA's episome persistence function would be expected to prevent or eliminate KSHV infection and provide effective therapy for KSHV associated malignancies. An understanding of the mechanisms underlying LANA mediated KSHV episome persistence is critical to enable the design of assays that screen small molecules which can inhibit these processes. Further, understanding the structure of the LANA DNA binding domain complexed with cognate DNA would enable rational drug design to interrupt LANA function. This work will investigate the mechanisms central to LANA's episome maintenance function. Proteins mediating LANA's association with chromosomes will be identified. The mechanisms underlying LANA mediated KSHV DNA replication will be investigated. Since the LANA C-terminus specifically binds KSHV DNA, investigations to solve the structure of this domain bound to cognate DNA will begin. Each of these aims investigates an area critical for LANA episome maintenance. Since episome persistence is central to latent infection and most tumor cells are latently infected, this work may lead to new strategies for prevention or treatment of KSHV associated malignancies.