Data and code associated with the publication: Disassembly of unstable RNA structures by an E. coli DEAD-box chaperone accelerates ribosome assembly

This dataset is related to a submitted manuscript describing how a general DEAD-box RNA chaperone facilitates the assembly of nascent rRNA into stable RNA-protein complexes, using single molecule total internal reflection fluorescence microscopy. Ribosome synthesis in bacteria is coupled with transcription of the pre-rRNA that must fold and assemble with 20 or more ribosomal proteins. In vitro, variable folding of the E. coli pre-16S rRNA during transcription delays stable addition of ribosomal protein uS4 that nucleates assembly of the 16S 5’ domain. Using single molecule fluorescence microscopy, we show that general unfolding by the DEAD-box protein CsdA (DeaD) strongly accelerates uS4 binding to nascent rRNA. Stable RNA structures resist unwinding by CsdA, which becomes less efficient as more ribosomal proteins join the complex. Monitoring single RNAs shows that dissociation of unstable complexes is followed by uS4 rebinding. Our results suggest that disassembly of nascent RNA-protein complexes by general chaperones drives cycles of reassembly, fueling the search for native-like complexes that resist unfolding and progress to mature ribosomes. (2024-08)