GENETIC AND BIOCHEMICAL STUDIES OF THE KSHV LANA GENE

KSHV LANA 基因的遗传学和生物化学研究

• 批准号: 6633407
• 负责人: Kenneth M Kaye
• 金额: $ 34.67万
• 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 2005-01-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6633407
• 关键词: Kaposi's sarcoma; clinical research; gel mobility shift assay; gene mutation; genetic library; human herpesvirus 8; human tissue; latent virus infection; plasmids; protein localization; tissue /cell culture; transfection; virus DNA; virus genetics; virus protein

Kaposi's sarcoma (KS) associated herpesvirus (KSHV or HHV8) is a recently discovered virus which is associated with malignancies in patients, especially those with AIDS. KSHV is found in KS, primary effusion lymphocytes (PELs) and multicentric Castleman's disease, a lymphoproliferative disorder. Similar to a closely related herpesvirus, Epstein-Barr virus, KSHV infection in malignancy and in PEL cell lines is primarily latent, with viral DNA persisting as an extra-chromosomal, circular episome. The latency associated nuclear antigen (LANA or LNA) is an 1162 amino acid KSHV protein expressed in a punctate distribution in the nuclei of latently infected cells. The serum of KSHV infected patients usually reacts with LANA and epidemiologic studies have often used this reactivity to determine KSHV seropositivity. The function of LANA is unknown. We now show that LANA colocalizes with KSHV episomes on cell chromosomes and LANA permits maintenance of KSHV episomes in cells. These results are consistent with a model in which LANA mediates segregation of episomes to daughter cells by tether episomes to cell chromosomes during mitosis. This project will test this model by determine if LANA colocalization with episomes on chromosomes is essential for episome maintenance. First, extensive mutational analysis of LANA will be performed to determine the LANA domains that mediate colocalization with chromosomes and with KSHV episomes. LANA mutants which cannot colocalize with chromosomes or episomes will be tested for loss of the ability to permit maintenance of KSHV episomes in cells. Next, the KSHV DNA element essential for LANA mediated episome maintenance will be defined. Experiments will then investigate the nature of the interaction between LANA and the essential KSHV DNA element.

卡波西肉瘤相关疱疹病毒（KSHV或HHV 8）是最近发现的一种病毒，与患者的恶性肿瘤，特别是艾滋病患者有关。KSHV存在于KS、原发性渗出淋巴细胞（PEL）和多中心Castleman病（一种淋巴组织增生性疾病）中。与密切相关的疱疹病毒Epstein-Barr病毒相似，恶性肿瘤和PEL细胞系中的KSHV感染主要是潜伏的，病毒DNA作为染色体外环状附加体持续存在。潜伏相关核抗原（拉娜或LNA）是在潜伏感染细胞的细胞核中以点状分布表达的1162个氨基酸的KSHV蛋白。KSHV感染患者的血清通常与拉娜反应，流行病学研究经常使用这种反应性来确定KSHV血清阳性。拉娜的功能未知。我们现在表明，拉娜与KSHV附加体共定位在细胞染色体上，并且拉娜允许KSHV附加体在细胞中维持。这些结果与其中拉娜通过在有丝分裂期间将附加体系在细胞染色体上来介导附加体向子细胞的分离的模型一致。本项目将通过确定拉娜与染色体上的附加体共定位是否对附加体维持至关重要来测试该模型。首先，将进行拉娜的广泛突变分析以确定介导与染色体和与KSHV附加体共定位的拉娜结构域。将测试不能与染色体或附加体共定位的拉娜突变体允许KSHV附加体在细胞中维持的能力的丧失。接下来，将定义拉娜介导的附加体维持所必需的KSHV DNA元件。然后实验将研究拉娜和KSHV基本DNA元件之间相互作用的性质。