SRNA OF PHAGE 029 ESSENTIAL FOR DNA PACKAGING

噬菌体 029 的 SRNA 对于 DNA 包装至关重要

• 批准号: 2185637
• 负责人: PEIXUAN GUO
• 金额: $ 12.23万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1996-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2185637
• 关键词: Bacillus subtilis; DNA; DNA footprinting; affinity labeling; autoradiography; bacterial virus; capsid; crosslink; intermolecular interaction; molecular cloning; mutant; nucleic acid probes; nucleic acid sequence; protein folding; protein sequence; radiotracer; small nuclear RNA; virus DNA; virus assembly

Our previous work has revealed that a novel small viral RNA (sRNA) has an essential role in bacteriophage phi29 DNA packaging. The sRNA is co- purified with the prohead but it is not found in the mature virion. Recently, involvement of RNA in DNA packaging has been demonstrated in phages lambda and D108, a Mmu-like phage. Similar to phi29, small RNAs without protein encoding capacity have also been found to be present in orthopoxvirus infected cells. If the involvement of sRNA in viral DNA packaging is a general phenomenon, then these sRNA molecules could be targets for anti-viral therapy. A major objective of this study is to elucidate fundamental mechanisms of the sRNA-protein interaction and the sRNA-DNA interaction during the maturation of the virus in order to understand the role of the sRNA in DNA packaging. Experiments will include: 1) isolation of phi29 DNA packaging intermediates; 2) analysis of the primary structure of the sRNA; 3) comparison of the secondary structure of the prohead-associated sRNA to that of the purified unassociated sRNA; 4) identification of sites for sRNA-DNA and sRNA-protein interactions; 5) determination of the active domain(s) of the sRNA; 6) calculation of the number of sRNA molecules required to package one DNA molecule; 7) studies on the rosette decamers resulting from sRNA-portal vertex interaction, which might provide information to support our model of the role of the sRNA in phi29 DNA packaging, and 8) studies on the antisense RNA against viral infections.

我们以前的工作表明，一种新型的小病毒RNA（SRNA）具有 噬菌体PHI29 DNA包装中的重要作用。 SRNA是 用Prohead纯化，但在成熟的病毒体中找不到。 最近，RNA参与DNA包装已在 Phages Lambda和D108，类似于MMU的噬菌体。 类似于phi29，小rnas 还发现没有蛋白质编码能力 正托病毒感染细胞。 如果SRNA参与病毒DNA 包装是一种一般现象，那么这些sRNA分子可能是 抗病毒疗法的靶标。 这项研究的主要目的是 阐明SRNA-蛋白质相互作用的基本机制和 病毒成熟期间的SRNA-DNA相互作用 了解SRNA在DNA包装中的作用。 实验会 包括：1）PHI29 DNA包装中间体的分离； 2）分析 SRNA的主要结构； 3）二级结构的比较 与纯化的非相关sRNA的亲头相关的SRNA的相关SRNA； 4）鉴定SRNA-DNA和SRNA-蛋白质相互作用的位点； 5） 确定SRNA的活动域； 6）计算 包装一个DNA分子所需的SRNA分子数量； 7）研究 在由sRNA-portal顶点相互作用产生的玫瑰花结demate上， 这可能会提供信息来支持我们的模型 PHI29 DNA包装中的SRNA和8）针对反义RNA的研究 病毒感染。