Emerging Quantitative Biochemical, Structural and Biophysical Methods to Study Ribosome and Protein-RNA Complex Assembly

• DOI: 10.20944/preprints202305.0503.v1
• Publication Date: 2023-05-09T00:35:46Z
• Authors: Kavan Gor; Olivier Duss
• Keywords: Ribosomal Proteins; Ribonucleoproteins; in vitro reconstitutions; protein–RNA interactions; RNA; RNA folding; Review; RNP assembly; ribosome assembly; Microbiology; Ribosomes; QR1-502; assembly intermediates

Ribosome assembly is one of the most fundamental processes in gene expression and has served as a playground to investigate the molecular mechanisms of how protein-RNA complexes (RNPs) assemble. The bacterial ribosome is composed of around 50 ribosomal proteins several of which are co-transcriptionally assembled on a ~4,500 nucleotides long pre-rRNA transcript that is further processed and modified during transcription, the entire process taking around 2 minutes in vivo and assisted by dozens of assembly factors. How this complex molecular process works so efficiently to produce an active ribosome has been investigated over decades and has resulted in the development of a plethora of novel approaches that can also be used to study the assembly of other RNPs. Here we review biochemical, structural and biophysical methods that have been developed and integrated to provide a detailed and quantitative understanding of this complex and intricate molecular process of assembly. We also discuss emerging cutting-edge approaches that could be used in the future to study how transcription, rRNA processing, cellular factors and the native cellular environment shape ribosome assembly and RNP assembly at large.