Genetic and Biochemical Studies of the KSHV LANA Gene

描述（由申请方提供）：卡波西肉瘤（KS）相关疱疹病毒（KSHV）与KS、原发性渗出性淋巴瘤（PEL）和多中心Castleman病在病因学上相关。这些疾病发生在艾滋病和其他患者中，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作为多拷贝、染色体外、环状附加体持续存在。几条证据支持这样一种模型，其中潜伏相关核抗原（拉娜）通过首先介导KSHV DNA的复制，然后将附加体束缚到染色体上以有效分离到后代细胞核来维持附加体。因此，中断拉娜的附加体持续功能的有效干预有望预防或消除KSHV感染，并为KSHV相关恶性肿瘤提供有效治疗。理解拉娜介导的KSHV附加体持久性的机制对于设计筛选可抑制这些过程的小分子的测定是至关重要的。此外，理解与同源DNA复合的拉娜DNA结合结构域的结构将使得合理的药物设计能够中断拉娜功能。这项工作将调查的机制中央拉娜的附加体维护功能。将鉴定介导拉娜与染色体结合的蛋白质。拉娜介导的KSHV DNA复制的机制将被研究。由于拉娜C-末端特异性结合KSHV DNA，因此将开始研究与同源DNA结合的该结构域的结构。这些目标中的每一个都研究了对拉娜附加体维持至关重要的区域。由于附加体持久性是潜伏感染的核心，大多数肿瘤细胞是潜伏感染，这项工作可能会导致新的策略，预防或治疗KSHV相关的恶性肿瘤。

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.