HHV-8 ORIGIN-DEPENDENT DNA REPLICATION

HHV-8 起源依赖性 DNA 复制

• 批准号: 6497968
• 负责人: GREGORY S PARI
• 金额: $ 25.97万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2005-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6497968
• 关键词: DNA replication; DNA replication origin; genetic mapping; human herpesvirus 8; virus genetics; virus replication

Kaposi's sarcoma herpesvirus (KSHV) or human herpesvirus type 8 (HHV-8) as it will be referred to in this proposal, is the likely cause of Kaposi's sarcoma (KS). Much of the data collected to date involves epidemiological evidence demonstrating that HHV-8 DNA sequences are present in a variety of lymphoid-derived cancers. For example, HHV-8 is implicated in multicentric Castleman's disease and body cavity-based large cell lymphomas. Despite intense research, little is known about HHV-8 lytic replication. Inspection of the HHV-8 nucleotide sequence does identify the candidate replication gene homologs to helicase- primase, polymerase and its processivity factor, and a single- stranded DNA binding protein. However none of these homologs have been shown by functional assay to be required for lytic replication. Now that the entire HHV-8 genome has been sequenced and putative ORFs have been identified, the next step is to identify the HHV-8 lytic origin of replication (oriLyt) and to elucidate those viral factors required for lytic DNA replication. To this end, a putative lytic origin of replication has been identified using a transient assay in BC3 cells. This proposal will: i) Characterize the putative lytic origin of replication for HHV-8, ii) identify the HHV-8 genes required for origin-dependent DNA replication, and iii) determine which genes participate directly in DNA replication and which play an ancillary role and characterize the HHV-8 initiator protein.

卡波西肉瘤疱疹病毒 (KSHV) 或本提案中将提及的人类疱疹病毒 8 型 (HHV-8) 可能是卡波西肉瘤 (KS) 的病因。 迄今为止收集的大部分数据涉及流行病学证据，证明 HHV-8 DNA 序列存在于多种淋巴源性癌症中。 例如，HHV-8 与多中心卡斯尔曼病和体腔大细胞淋巴瘤有关。尽管进行了大量研究，但人们对 HHV-8 裂解复制知之甚少。 HHV-8核苷酸序列的检查确实鉴定了与解旋酶引物酶、聚合酶及其持续因子以及单链DNA结合蛋白同源的候选复制基因。 然而，这些同源物均未通过功能测定显示为裂解复制所必需的。 现在，整个 HHV-8 基因组已完成测序，推定的 ORF 也已确定，下一步是确定 HHV-8 裂解性复制起点 (oriLyt) 并阐明裂解性 DNA 复制所需的病毒因子。为此，在 BC3 细胞中使用瞬时测定法鉴定了假定的裂解性复制起点。该提案将：i) 表征 HHV-8 假定的裂解性复制起点，ii) 识别起点依赖性 DNA 复制所需的 HHV-8 基因，以及 iii) 确定哪些基因直接参与 DNA 复制，哪些基因起辅助作用。 HHV-8 起始蛋白的作用和表征。