Conformational activation of ribosome recycling by intra- and inter-molecular dynamics of RRF
| Autor: | Huigang Shi, Xinzheng Zhang, Yan Qin, Yan-Zhong Guan, Dejiu Zhang, Yanhui Chu, Yong Zhang, Jizhong Lou, Xintao Cao, Guangtao Song, Benjin Xu, Zengrui Liu, Ran Guo |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Ribosomal Proteins
Cryo-electron microscopy Protein subunit Ribosome Recycling Factor 02 engineering and technology Molecular Dynamics Simulation Biochemistry Ribosome 03 medical and health sciences Molecular dynamics Structural Biology Molecular Biology 030304 developmental biology 0303 health sciences biology Chemistry Escherichia coli Proteins Translation (biology) General Medicine Single-molecule FRET Peptide Chain Termination Translational 021001 nanoscience & nanotechnology Single-molecule experiment biology.protein Biophysics 0210 nano-technology Ribosomes |
| Zdroj: | International Journal of Biological Macromolecules. 160:1212-1219 |
| ISSN: | 0141-8130 |
| DOI: | 10.1016/j.ijbiomac.2020.05.254 |
| Popis: | Ribosome recycling is the final step of the cyclic process of translation, where the post-termination complex (PoTC) is disassembled by the concerted action of ribosome recycling factor (RRF) and elongation factor G (EF-G) in the sub-second time range. Since, however, both the RRF and PoTC display highly dynamic action during this process, it is difficult to assess the molecular details of the interactions between the factors and the ribosome that are essential for rapid subunit separation. Here we characterized the molecular dynamics of RRF and PoTC by combined use of molecular dynamics simulations, single molecule fluorescence detection and single-particle cryo-EM analysis, with time resolutions in the sub-millisecond to minute range. We found that RRF displays two-layer dynamics: intra- and inter-molecular dynamics during ribosome splitting. The intra-molecular dynamics exhibits two different configurations of RRF: 'bent' and 'extended'. A single-site mutant of RRF increases its propensity to the 'extended' conformation and leads to a higher binding affinity of RRF to the PoTC. The inter-molecular dynamics between RRF and EF-G in the PoTC reveals that the domain IV of EF-G pushes against the domain II of RRF, triggering the disruption of the major inter-subunit bridge B2a, and catalyzes the splitting. |
| Databáze: | OpenAIRE |
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