Abstract
Upon encountering a stop codon on mRNA, polypeptide synthesis on the ribosome is terminated by release factors, and the ribosome complex, still bound with mRNA and P-site-bound tRNA (post-termination complex, PostTC), is split into ribosomal subunits, ready for a new round of translational initiation. Separation of post-termination ribosomes into subunits, or "ribosome recycling, " is promoted by the joint action of ribosome-recycling factor (RRF) and elongation factor G (EF-G) in a guanosine triphosphate (GTP) hydrolysisdependent manner. Here we used a mixing-spraying-based method of timeresolved cryo-electron microscopy (cryo-EM) to visualize the short-lived intermediates of the recycling process. The two complexes that contain (1) both RRF and EF-G bound to the PostTC or (2) deacylated tRNA bound to the 30S subunit are of particular interest. Our observations of the native form of these complexes demonstrate the strong potential of time-resolved cryo-EM for visualizing previously unobservable transient structures.
| Original language | English |
|---|---|
| Title of host publication | Novel Developments in Cryo-EM of Biological Molecules |
| Subtitle of host publication | Resolution in Time and State Space |
| Publisher | Taylor and Francis |
| Pages | 367-393 |
| Number of pages | 27 |
| ISBN (Electronic) | 9781000989441 |
| ISBN (Print) | 9789814968768 |
| State | Published - Oct 6 2023 |