DNA Replication and Gene Expression of Chlorella Viruses

小球藻病毒的 DNA 复制和基因表达

• 批准号: 7105625
• 负责人: JAMES L VAN ETTEN
• 金额: $ 30.65万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-09-28 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7105625
• 关键词: Chlorophyta; DNA replication; DNA virus; Xenopus oocyte; capsid; enzyme activity; fatty acylation; gene expression; glycosylation; glycosyltransferase; host organism interaction; immunoprecipitation; mass spectrometry; myristates; plant virus; posttranslational modifications; potassium channel; protein localization; protein protein interaction; protein structure function; site directed mutagenesis; virus genetics; virus protein

DESCRIPTION (provided by applicant): The large (>330 kb) dsDNA, plaque-forming chlorella viruses have provided new and unexpected gene products for the past 20 years, including several medically relevant proteins. The objectives of this proposal are a step toward the long-term goal of understanding the structural and functional aspects of these viruses, as well as their associated genes and gene products. Many chlorella virus proteins are the smallest representatives of their functional class. Consequently, some of these proteins are the subject of intense physical and biochemical investigation, e.g. a type II DNA topoisomerase and a potassium ion channel protein Kcv. This proposal focuses on two important problems associated with virus entry into the cell and maturation of progeny viruses. The specific aims of this proposal are to: i) examine kcv genes and their corresponding proteins from a broad range of chlorella viruses as an alternate approach for exploring structure/function relationships in potassium ion channels and ii) evaluate unusual virus-encoded posttranslational modifications of the virus major capsid protein, Vp54. In addition to characterizing the seven putative virus-encoded glycosyltransferases, the structures of the six glycans attached to the major capsid protein will be determined. The viral proteins will be investigated at physiological, molecular biological and biochemical levels, including structural analyses. These studies are expected to reveal new concepts in intra- and inter-molecular interactions of potassium ion channels, and also novel mechanisms for protein glycosylation and myristoylation. Phylogenetic analyses indicate the genetically diverse but morphologically indistinguishable chlorella viruses have a long evolutionary history, possibly dating back to the time eukaryotes and prokaryotes separated. Thus, chlorella viruses are an important model for studying large nucleocytoplasmic dsDNA viruses because they have evolutionary ties to important mammalian viruses, such as the pox viruses and the quarantined African swine fever virus.

描述（由申请人提供）：大型（> 330 kb）dsDNA、形成空斑的小球藻病毒在过去20年中提供了新的和意想不到的基因产物，包括几种医学相关的蛋白质。 该提案的目标是朝着了解这些病毒的结构和功能方面及其相关基因和基因产物的长期目标迈出一步。 许多小球藻病毒蛋白是其功能类别的最小代表。 因此，其中一些蛋白质是深入的物理和生化研究的主题，例如。 II 型 DNA 拓扑异构酶和钾离子通道蛋白 Kcv。该提案重点关注与病毒进入细胞和子代病毒成熟相关的两个重要问题。 该提案的具体目标是：i) 检查来自多种小球藻病毒的 kcv 基因及其相应的蛋白质，作为探索钾离子通道结构/功能关系的替代方法；ii) 评估不寻常的病毒编码的翻译后修饰病毒主要衣壳蛋白，Vp54。 除了表征七种假定的病毒编码糖基转移酶外，还将确定与主要衣壳蛋白相连的六种聚糖的结构。 病毒蛋白将在生理、分子生物学和生化水平上进行研究，包括结构分析。 这些研究有望揭示钾离子通道分子内和分子间相互作用的新概念，以及蛋白质糖基化和肉豆蔻酰化的新机制。 系统发育分析表明，遗传多样性但形态上无法区分的小球藻病毒具有悠久的进化历史，可能可以追溯到真核生物和原核生物分离的时代。 因此，小球藻病毒是研究大型核质双链DNA病毒的重要模型，因为它们与重要的哺乳动物病毒（例如痘病毒和检疫的非洲猪瘟病毒）具有进化联系。