EMD-9530
Cryo-EM structure of the model post-termination complex (PoTC) in the rotated state
EMD-9530
Single-particle8.5 Å
Deposition: 30/07/2016Map released: 07/06/2017
Last modified: 12/10/2022
Sample Organism:
Escherichia coli
Sample: Model post-termination complex (PoTC)
Deposition Authors: Iwakura N, Yokoyama T
,
Quaglia F,
Mitsuoka K ,
Mio K ,
Shigematsu H ,
Shirouzu M,
Kaji A,
Kaji H
Sample: Model post-termination complex (PoTC)
Deposition Authors: Iwakura N, Yokoyama T
,
Quaglia F,
Mitsuoka K ,
Mio K ,
Shigematsu H ,
Shirouzu M,
Kaji A,
Kaji H
Chemical and structural characterization of a model Post-Termination Complex (PoTC) for the ribosome recycling reaction: Evidence for the release of the mRNA by RRF and EF-G.
Iwakura N,
Yokoyama T ,
Quaglia F,
Mitsuoka K ,
Mio K ,
Shigematsu H ,
Shirouzu M,
Kaji A,
Kaji H
(2017) PLoS One , 12 , e0177972 - e0177972
,
Quaglia F,
Mitsuoka K ,
Mio K ,
Shigematsu H ,
Shirouzu M,
Kaji A,
Kaji H
(2017) PLoS One , 12 , e0177972 - e0177972
Abstract:
A model Post-Termination Complex (PoTC) used for the discovery of Ribosome Recycling Factor (RRF) was purified and characterized by cryo-electron microscopic analysis and biochemical methods. We established that the model PoTC has mostly one tRNA, at the P/E or P/P position, together with one mRNA. The structural studies were supported by the biochemical measurement of bound tRNA and mRNA. Using this substrate, we establish that the release of tRNA, release of mRNA and splitting of ribosomal subunits occur during the recycling reaction. Order of these events is tRNA release first followed by mRNA release and splitting almost simultaneously. Moreover, we demonstrate that IF3 is not involved in any of the recycling reactions but simply prevents the re-association of split ribosomal subunits. Our finding demonstrates that the important function of RRF includes the release of mRNA, which is often missed by the use of a short ORF with the Shine-Dalgarno sequence near the termination site.
A model Post-Termination Complex (PoTC) used for the discovery of Ribosome Recycling Factor (RRF) was purified and characterized by cryo-electron microscopic analysis and biochemical methods. We established that the model PoTC has mostly one tRNA, at the P/E or P/P position, together with one mRNA. The structural studies were supported by the biochemical measurement of bound tRNA and mRNA. Using this substrate, we establish that the release of tRNA, release of mRNA and splitting of ribosomal subunits occur during the recycling reaction. Order of these events is tRNA release first followed by mRNA release and splitting almost simultaneously. Moreover, we demonstrate that IF3 is not involved in any of the recycling reactions but simply prevents the re-association of split ribosomal subunits. Our finding demonstrates that the important function of RRF includes the release of mRNA, which is often missed by the use of a short ORF with the Shine-Dalgarno sequence near the termination site.