ASSEMBLY OF THE 30S RIBOSOMAL SUBUNIT

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 A）细菌30 S核糖体由1542个核苷酸的16 S核糖体RNA和20个小核糖体蛋白组成。 在蛋白质合成过程中，它负责与50 S亚基相关的mRNA解码。 E.大肠杆菌可以从纯化的组分在体外重建。 已经开发了一种组装图，其包含平行蛋白质结合和顺序蛋白质结合作为机械特征。 30 S组装的新图景是，它是组装途径的统计分布，但在组装景观中存在有利的路线。 与蛋白质子集结合的中间体在组装过程中积累，但这些中间体最终聚集在最终完成的亚基上。 我们感兴趣的是通过在体外重建过程中观察大量的核糖体颗粒来测量中间体群体的时间依赖性。 B）30 S亚基是分子组装体自发形成的范例。 无论是从结构上还是从装配的角度来看，它都是最具特色的大型系统之一。 了解30 S组装的详细机制将为涉及大量组件的许多类别的分子机器的组装提供见解。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A) The bacterial 30S ribosome is composed of a 1542 nucleotide 16S ribosomal RNA and 20 small ribosomal proteins. It is responsible for mRNA decoding in association with the 50S subunit during protein synthesis. The 30S subunit from E. coli can be reconsituted in vitro from purified components. An assembly map has been developed that contains both parallel protein binding and sequential protein binding as mechanistic features. The emerging picture of 30S assembly is that it is a statistical distribution of assembly pathways, but that there are favored routes over the assembly landscape. Intermediates with subsets of proteins bound accumulate during the assembly, but these intermediates eventually converge on the final completed subunit. We are interested in measuring the time dependence of the populations of intermediates by visualizing large number of ribosomal particles during in vitro reconstitution. B) The 30S subunit is a paradigm for spontaneous formation of a molecular assembly. It is one of the best characterized large systems both structurally and from the standpoint of assembly. Understanding the detailed mechanism of 30S assembly will provide insights into the assembly of many classes of molecular machines involving large numbers of components.