HHV-8 ORIGIN-DEPENDENT DNA REPLICATION

HHV-8 起源依赖性 DNA 复制

• 批准号: 6628442
• 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/6628442
• 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.

Kaposi的肉瘤疱疹病毒（KSHV）或人类疱疹病毒8型（HHV-8）将在此提案中提到，这可能是Kaposi的肉瘤（KS）的原因。 迄今为止收集的许多数据涉及流行病学证据，表明HHV-8 DNA序列存在于多种淋巴样衍生的癌症中。 例如，HHV-8与多中心Castleman病和基于体腔的大细胞淋巴瘤有关。尽管进行了激烈的研究，但对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候选复制基因与解旋酶 - 原始酶，聚合酶及其加工性因子以及单链DNA结合蛋白的同源物。 但是，这些同源物没有通过功能分析表明裂解复制所需的。 既然已经对整个HHV-8基因组进行了测序并已经确定了推定的ORF，下一步是识别复制的HHV-8裂解起源（Orilyt），并阐明裂解DNA复制所需的病毒因子。为此，使用BC3细胞中的瞬态测定法确定了重复的假定裂解起源。该提议将：i）表征HHV-8复制的假定裂解起源，ii）确定依赖原点依赖性DNA复制所需的HHV-8基因，iii）确定哪些基因直接参与DNA复制，哪些基因参与了辅助作用并扮演辅助作用并表征了HHV-8启动器蛋白。