STRUCTURE OF VIRAL DNA PACKAGING MACHINE & PACKAGING INTERMEDIATES

病毒DNA包装机结构

• 批准号: 6308932
• 负责人: DWIGHT ANDERSON
• 金额: $ 0.97万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6308932
• 关键词: biomedical resource; enzymes; genetics; genome; growth /development; nucleic acids; virus

Bacteriophage 1029 of A subtilis is an excellent model for studies of viral DNA packaging. The in vitro packaging system of (029 DNA is as efficient as in vivo assembly. Models of DNA packaging can be tested and the structum of components can be determined. 1029 DNA with covalently bound gene product 3 at each terminus (DNA-gp3) is translocated in vitro into a preformed prvteir~shell (prohead) by a machine that includes the prohead'portal vertex Oead-tail comector), the 174-base 4029-encoded prohead RNA.and the ATPase gp16. The prohead, connector, pRNA and gp16 are all overproduced from cloned genes permitting the structure of these components in sequential interactions with DNA-gp3 to be studied. Supercoiled pBR322 DNA wraps around the outside of the isolated 4~29 connector. This is hypothesized to reflect the initial phase of DNA packaging. In this model, proheads should also bind supercoiled DNA and 40 DNA-gp3 should be supercoiled as a prerequisite for packaging. , An RNA pseudoknot, an intramolecular tertiary interaction, in the pRNA was inferred and then confirmed by genetic mutations. Two mutants, neither of winch can make the mtramolecular pseudoknot, can complement each other in the prohead packaging assay indicating they might make an intermolecular psuedoknot. Masses of different pRNAs were measured in the STEM to try to confirm this. It is planned to look at pRNA-connector complexes in the STEM.

枯草杆菌噬菌体1029是一个很好的研究模型 病毒DNA包装。(029)DNA的体外包装系统 与体内组装一样高效。DNA包装的模型可以 经过测试，可以确定部件的结构。1029 DNA 在每个末端带有共价结合的基因产物3(DNA-GP3)是 通过体外移植到预制的Prvteir~壳(Prohead)中 包括前部‘门户顶点Oead-Tail角的机器)， 174碱基4029编码的前导RNA和ATPase gp16。这个 Prohead、Connector、PRNA和gp16都是从克隆的 允许这些成分的结构按顺序排列的基因 与DNA-GP3的相互作用有待研究。超螺旋pBR322 DNA包装 在隔离的4~29连接器的外侧周围。这是 假设反映了DNA包装的初始阶段。在这 模型，探针也应该结合超螺旋DNA和40 DNA-GP3应该 作为包装的前提条件进行超级卷曲。，一个RNA伪结， 推测了PRNA中的分子内三级相互作用。 然后通过基因突变来确认。两个突变体，都不是Winch 可以使分子间的假结，可以互补中 前置包装化验表明他们可能会 分子间的假结。不同RNA的质量被测量在 STEM试图证实这一点。它计划要看一看 STEM中的PRNA-连接器复合体。