In this study, cryo-electron microscopy and subclassification were used to identify intermediates in the assembly of the 50S ribosomal subunit from E. coli. The study reveals a complete assembly map for all RNA and protein components, and shows that assembly generally proceeds co-transcriptionally.
Assembly of ribosomes in bacteria is highly efficient, taking similar to 2-3 min, but this makes the abundance of assembly intermediates very low, which is a challenge formechanistic understanding. Genetic perturbations of the assembly process create bottlenecks where intermediates accumulate, facilitating structural characterization. We use cryo-electron microscopy, with iterative subclassification to identify intermediates in the assembly of the 50S ribosomal subunit from E. coli. The analysis of the ensemble of intermediates that spans the entire biogenesis pathway for the 50 S subunit was facilitated by a dimensionality reduction and cluster picking approach using PCA-UMAPHDBSCAN. The identity of the cooperative folding units in the RNA with associated proteins is revealed, and the hierarchy of these units reveals a complete assembly map for all RNA and protein components. The assembly generally proceeds co-transcriptionally, with some flexibility in the landscape to ensure efficiency for this central cellular process under a variety of growth conditions.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据