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Title | Structural Visualization of the Formation and Activation of the 50S Ribosomal Subunit during In Vitro Reconstitution. |
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Journal, issue, pages | Mol Cell, Vol. 70, Issue 5, Page 881-893.e3, Year 2018 |
Publish date | Jun 7, 2018 |
Authors | Rainer Nikolay / Tarek Hilal / Bo Qin / Thorsten Mielke / Jörg Bürger / Justus Loerke / Kathrin Textoris-Taube / Knud H Nierhaus / Christian M T Spahn / |
PubMed Abstract | The assembly of ribosomal subunits is an essential prerequisite for protein biosynthesis in all domains of life. Although biochemical and biophysical approaches have advanced our understanding of ...The assembly of ribosomal subunits is an essential prerequisite for protein biosynthesis in all domains of life. Although biochemical and biophysical approaches have advanced our understanding of ribosome assembly, our mechanistic comprehension of this process is still limited. Here, we perform an in vitro reconstitution of the Escherichia coli 50S ribosomal subunit. Late reconstitution products were subjected to high-resolution cryo-electron microscopy and multiparticle refinement analysis to reconstruct five distinct precursors of the 50S subunit with 4.3-3.8 Å resolution. These assembly intermediates define a progressive maturation pathway culminating in a late assembly particle, whose structure is more than 96% identical to a mature 50S subunit. Our structures monitor the formation and stabilization of structural elements in a nascent particle in unprecedented detail and identify the maturation of the rRNA-based peptidyl transferase center as the final critical step along the 50S assembly pathway. |
External links | Mol Cell / PubMed:29883607 |
Methods | EM (single particle) |
Resolution | 3.8 - 4.3 Å |
Structure data | EMDB-4378, PDB-6gbz: EMDB-4379, PDB-6gc0: EMDB-4380, PDB-6gc4: EMDB-4381, PDB-6gc6: |
Chemicals | ChemComp-ZN: ChemComp-MG: ChemComp-HOH: |
Source |
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Keywords | RIBOSOME / assembly precursor 48S 50S biogenesis in vitro reconstitution |