STRUCTURAL DETERMINATION OF PF-8 IN-COMPLEX WITH DNA

PF-8 与 DNA 复合物的结构测定

• 批准号: 8361731
• 负责人: JAMES HOGLE
• 金额: $ 1.46万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361731
• 关键词: Binding; Complex; Cytomegalovirus; DNA; DNA Sequence Rearrangement; DNA biosynthesis; DNA-Directed DNA Polymerase; Data; Funding; Grant; Head; Herpesviridae; Herpesvirus 1; Human; Human Herpesvirus 8; Kaposi Sarcoma; Lymphoma; Lymphoproliferative Disorders; Modeling; Multicentric Angiofollicular Lymphoid Hyperplasia; National Center for Research Resources; Polymerase; Principal Investigator; Proliferating Cell Nuclear Antigen; Research; Research Infrastructure; Resources; Site; Source; Structure; United States National Institutes of Health; Viral; base; comparative; cost; dimer; effusion; falls; gammaherpesvirus; insight; monomer; sarcoma; structural biology

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Human herpesvirus-8 (HHV-8), also known as Karposi's sarcoma-associated human herpesvirus (KSHV), is a gammaherpesvirus that is the causative agent of various lymphoproliferative disorders including Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. KSHV encodes the viral processivity factor, PF-8, which binds to DNA and is essential for maintaining contact between the viral polymerase Pol-8 and DNA during DNA synthesis. The crystal structures of a processivity factor from the alpha herpesvirus, HSV-1, and the beta herpesvirus, HCMV, revealed structures similar to that of the eukaryotic processivity factor, proliferating cell nuclear antigen (PCNA), except in their quaternary rearrangement. PCNA forms a trimeric ring around DNA while the processivity factors from HSV-1 and HCMV are a monomer and dimer respectively and do not require a clamp loader or ATP in order to be loaded onto DNA. We have determined the crystal structure of PF-8 to 2.8¿, and show that PF-8 forms a head-to-head homodimer that appears to fall, functionally, somewhere between the structures of UL42 and UL44. Based on the structural data, we are able to identify possible sites for interactions with DNA and polymerase, to propose a model for the simultaneous interaction of all three components of the complex. In addition, the availability of crystal structures for all three herpesvirus classes provides new insights into comparative structure and function.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 人类疱疹病毒 8 (HHV-8) 也称为卡波西肉瘤相关人类疱疹病毒 (KSHV)，是一种伽马疱疹病毒，是多种淋巴组织增生性疾病的病原体，包括卡波西肉瘤、原发性渗出性淋巴瘤和多中心卡斯尔曼病。 KSHV 编码病毒持续生长因子 PF-8，该因子与 DNA 结合，对于在 DNA 合成过程中维持病毒聚合酶 Pol-8 和 DNA 之间的接触至关重要。来自α疱疹病毒HSV-1和β疱疹病毒HCMV的持续生长因子的晶体结构揭示了与真核持续生长因子增殖细胞核抗原（PCNA）相似的结构，除了它们的四级重排之外。 PCNA 在 DNA 周围形成三聚环，而来自 HSV-1 和 HCMV 的持续合成因子分别是单体和二聚体，不需要夹具加载器或 ATP 即可加载到 DNA 上。我们已经确定了 PF-8 的晶体结构为 2.8¿，并表明 PF-8 形成了头对头的同二聚体，其功能似乎介于 UL42 和 UL44 的结构之间。根据结构数据，我们能够识别与 DNA 和聚合酶相互作用的可能位点，从而提出复合物所有三个组分同时相互作用的模型。此外，所有三种疱疹病毒类别的晶体结构的可用性为比较结构和功能提供了新的见解。