The ribosome synchronizes folding and assembly to promote oligomeric protein biogenesis
Natural proteins are structurally diverse and often form intricate multidomain, oligomeric architectures. This presents a prima facie challenge to cellular homeostasis, as topologically complex proteins seldom refold efficiently in vitro. How cells overcome sequence-intrinsic folding limitations to optimize protein biogenesis is incompletely understood. Here, we show that efficient folding and assembly of a model 5-domain homotetramer is obligatorily coupled to its synthesis on the ribosome, and define the underlying mechanisms. During refolding of full-length protein from denaturant, maturation of the catalytic TIM barrel domain is frustrated. Assembly outpaces monomer folding, and non-native oligomers accumulate. The ribosome directs the order of folding events and specifies the pathway of oligomer assembly. Efficient de novo folding is characterised by segmental folding of the TIM barrel, shaped by binding of a nascent amphipathic helix to a cryptic pocket on the ribosome surface. Homomer assembly initiates cotranslationally via recruitment of a full-length subunit to the nascent polypeptide, and the failure to do so results in misassembly. Our results reveal how the ribosome can dictate the timing of folding and assembly to enable efficient biogenesis of a topologically complex protein. Overall design: Probing cotranslational assembly of beta-galactosidase by selective ribosome profiling (https://doi.org/10.1038/s41586-018-0462-y) and disome selective profiling (DOI: 10.1126/science.abc7151).