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Structure paper

TitleStructural Visualization of the Formation and Activation of the 50S Ribosomal Subunit during In Vitro Reconstitution.
Journal, issue, pagesMol Cell, Vol. 70, Issue 5, Page 881-893.e3, Year 2018
Publish dateJun 7, 2018
AuthorsRainer Nikolay / Tarek Hilal / Bo Qin / Thorsten Mielke / Jörg Bürger / Justus Loerke / Kathrin Textoris-Taube / Knud H Nierhaus / Christian M T Spahn /
PubMed AbstractThe 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 linksMol Cell / PubMed:29883607
MethodsEM (single particle)
Resolution3.8 - 4.3 Å
Structure data

4378

6gbz

EMDB-4378, PDB-6gbz:
50S ribosomal subunit assembly intermediate state 5
Method: EM (single particle) / Resolution: 3.8 Å

4379

6gc0

EMDB-4379, PDB-6gc0:
50S ribosomal subunit assembly intermediate state 4
Method: EM (single particle) / Resolution: 3.8 Å

4380

6gc4

EMDB-4380, PDB-6gc4:
50S ribosomal subunit assembly intermediate state 3
Method: EM (single particle) / Resolution: 4.3 Å

4381

6gc6

EMDB-4381, PDB-6gc6:
50S ribosomal subunit assembly intermediate state 2
Method: EM (single particle) / Resolution: 4.3 Å

4382

6gc7

EMDB-4382, PDB-6gc7:
50S ribosomal subunit assembly intermediate state 1
Method: EM (single particle) / Resolution: 4.3 Å

4383

6gc8

EMDB-4383, PDB-6gc8:
50S ribosomal subunit assembly intermediate - 50S rec*
Method: EM (single particle) / Resolution: 3.8 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-HOH:
WATER / Water

Source
  • escherichia coli (E. coli)
  • escherichia coli k-12 (bacteria)
  • escherichia coli (strain k12) (bacteria)
KeywordsRIBOSOME / assembly precursor 48S 50S biogenesis in vitro reconstitution

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