Key Intermediates in Ribosome Recycling Visualized by Time-Resolved Cryoelectron Microscopy
2016 (English)In: Structure, ISSN 0969-2126, E-ISSN 1878-4186, Vol. 24, no 12, 2092-2101 p.Article in journal (Refereed) Published
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) hydrolysis-dependent manner. Here we used a mixing-spraying-based method of time-resolved 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.
Place, publisher, year, edition, pages
2016. Vol. 24, no 12, 2092-2101 p.
Cell and Molecular Biology
IdentifiersURN: urn:nbn:se:uu:diva-316969DOI: 10.1016/j.str.2016.09.014ISI: 000393211300009PubMedID: 27818103OAI: oai:DiVA.org:uu-316969DiVA: diva2:1079450
FunderNIH (National Institute of Health), R01 GM55440 GM29169Swedish Research Council, 2015-04682Knut and Alice Wallenberg Foundation