Domain consolidation in Bacterial 50S assembly revealed by Anti-Sense Oligonucleotide Probing.

ABSTRACT: Investigating the intricate and rapid folding kinetics of large RNA-protein complexes (RNPs), like the bacterial ribosome, remains a formidable challenge in structural biology. Previous genetic approaches to probe assembly have focused on modulating the expression of either r-proteins or assembly factors. Here, anti-sense oligonucleotides (ASOs) were used to disrupt native RNA/RNA and RNA/protein interactions, in order to generate novel folding intermediates. In an in vitro co-transcriptional assembly assay, 8 assembly inhibitor ASOs were identified. Using cryo-electron microscopy, 38 new intermediate structures were determined resulting from the specific inhibitions. In particular a novel intermediate class provided compelling evidence of independent rRNA domain folding before prop

SUBMITTER: Sheng K 

PROVIDER: S-EPMC11100757 | biostudies-literature | 2024 May

REPOSITORIES: biostudies-literature