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  Structural Visualization of the Formation and Activation of the 50S Ribosomal Subunit during In Vitro Reconstitution

Nikolay, R., Hilal, T., Qin, B., Mielke, T., Bürger, J., Loerke, J., et al. (2018). Structural Visualization of the Formation and Activation of the 50S Ribosomal Subunit during In Vitro Reconstitution. Molecular Cell, 70(5): e3, pp. 881-893. doi:10.1016/j.molcel.2018.05.003.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0003-55FE-0 Version Permalink: http://hdl.handle.net/21.11116/0000-0003-55FF-F
Genre: Journal Article

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 Creators:
Nikolay, Rainer , Author
Hilal, Tarek , Author
Qin, Bo, Author
Mielke, Thorsten1, Author              
Bürger, Jörg1, Author              
Loerke, Justus , Author
Textoris-Taube, Kathrin , Author
Nierhaus, Knud H. , Author
Spahn, Christian M.T. , Author
Affiliations:
1Microscopy and Cryo-Electron Microscopy (Head: Thorsten Mielke), Scientific Service (Head: Christoph Krukenkamp), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479668              

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Free keywords: 50S in vitro reconstitution; 50S subunit; cryo-EM map; cryo-electron microscopy; large ribosomal subunit; ribosome; ribosome assembly; ribosome biogenesis; structure
 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 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.

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Language(s): eng - English
 Dates: 2018-06-07
 Publication Status: Published online
 Pages: -
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 Table of Contents: -
 Rev. Method: -
 Identifiers: DOI: 10.1016/j.molcel.2018.05.003
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Title: Molecular Cell
Source Genre: Journal
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Publ. Info: Cambridge, Mass. : Cell Press
Pages: 13 Volume / Issue: 70 (5) Sequence Number: e3 Start / End Page: 881 - 893 Identifier: ISSN: 1097-2765
CoNE: https://pure.mpg.de/cone/journals/resource/954925610929