Packaging of the Segmented Genome of Bacteriophage Phi6

噬菌体 Phi6 分段基因组的包装

• 批准号: 7254096
• 负责人: LEONARD E MINDICH
• 金额: $ 42.68万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-01-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7254096
• 关键词: Animals; Bacteriophages; Binding; Binding Sites; Complex; Condition; Cryoelectron Microscopy; Crystallization; Cystoviridae; Disease; Double Stranded RNA Virus; Double-Stranded RNA; Elements; Epitopes; Family; Family member; Future; Genetic; Genome; Genomic Segment; Genomics; Goals; Human; In Vitro; Integration Host Factors; Investigation; Libraries; Link; Location; Maps; Modeling; Molecular Conformation; Molecular Structure; Motor; Numbers; Peptide Sequence Determination; Peptides; Phage Display; Plants; Protein Region; Proteins; RNA; RNA Binding; Reagent; Reoviridae; Series; Site; Specificity; Staging; Structural Protein; Structure; Surface; System; Transcript; Viral; Viral Genes; Work; comparative; crosslink; image reconstruction; in vivo; member; nucleoside triphosphatase; programs; research study

DESCRIPTION (provided by applicant): Bacteriophages of the family cystoviridae are unique in that they have genomes of 3 segments of double-stranded RNA packaged precisely within a preformed procapsid. Phi6 is the first member of this family and has been studied extensively. A model has been proposed for the mechanism of precise genomic packaging in phi6. The model is strongly supported by the confirmation of predicted results in both in vitro and in vivo packaging. The packaging involves the serially dependent transport of plus strands s, m and I into preformed procapsids with specificity determined by changes in the binding sites on the outside of P1, the major structural protein of the procapsid. The goal of this project is to determine the interaction of the RNA molecules with specific parts of protein P1; to identify the specific elements involved in the changing of the RNA binding sites as the packaging program procedes. This proposal is for the investigation of the location of the binding sites, their amino acid sequences and the determination of the interaction between the RNA and the binding sites. A major question posed is whether the binding sites for the different RNA molecules are physically separate from each other or whether they are overlapping and involve the same regions of protein P1. Approaches utilized in this project include the cross-linking of specific RNA molecules to their binding sites and the use of phage display libraries to locate binding sites. Collaborative structural studies are also proposed in which cores that have packaged different amounts of RNA will be examined by cryoelectron microscopy to determine their changes in conformation. This study also includes a comparative investigation of genomic packaging in the distantly related bacteriophage phi8, which seems to follow the general rules determined for phi6 but with less stringency and with a number of additional features, particularly the participation of host factors and viral genes not found in other members of the cystoviridae. The cystoviridae are the only segmented dsRNA viruses whose mode of packaging is understood. There is no current understanding of the genomic packaging of the reoviridae, segmented dsRNA viruses that cause disease in plants and animals, including humans.

描述（由申请人提供）：猫科动物科的噬菌体是独一无二的，因为它们具有3个片段的基因组，该基因组为预先形成的procapsid中精确包装的双链RNA。 Phi6是该家族的第一个成员，并且已经进行了广泛的研究。已经提出了一个模型，以实现PHI6中精确基因组包装的机理。该模型在体外和体内包装中都得到了预测的结果的强烈支持。该包装涉及加链s，m和i的串行依赖转运，其特异性由p1外部的结合位点的变化确定，这是procapsid的主要结构蛋白。该项目的目的是确定RNA分子与蛋白质P1的特定部分的相互作用。确定RNA绑定位点更改的特定元素作为包装程序程序。该建议是用于研究结合位点的位置，其氨基酸序列以及RNA与结合位点之间相互作用的测定。提出的一个主要问题是，不同RNA分子的结合位点是否在物理上是彼此分离的，还是重叠并涉及蛋白质P1的相同区域。该项目中使用的方法包括特定的RNA分子与其结合位点的交联，以及使用噬菌体显示库来定位结合位点。还提出了协作结构研究，其中将通过冷冻电子显微镜检查包装不同量的RNA的核心，以确定它们的构象变化。这项研究还包括对远距离相关的噬菌体PHI8中基因组包装的比较研究，该研究似乎遵循了针对PHI6确定的一般规则，但具有较少的严格性和许多其他特征，尤其是在Cystoviridae其他成员中发现的宿主因素和病毒基因的参与。半胱氨酸杆菌是唯一了解包装方式的分段DSRNA病毒。目前尚无对依伏迪科，分割的DSRNA病毒的基因组包装的理解，这些病毒在包括人类在内的动植物中引起疾病​​。