8RCM

Escherichia coli paused disome complex (Non-rotated disome interface class 2)

This is a non-PDB format compatible entry.

Summary for 8RCM

• Entry DOI: 10.2210/pdb8rcm/pdb
• Related: 8PEG 8PKL 8R3V
• EMDB information: 17631 17743 19055
• Descriptor: 50S ribosomal protein L28, Small ribosomal subunit protein uS4, Small ribosomal subunit protein uS5, ... (49 entities in total)
• Functional Keywords: ribosome, polysome, tranlsation, elongation, pausing, disome, collision, pure system
• Biological source: Escherichia coli; More
• Total number of polymer chains: 65
• Total formula weight: 3882660.00

Authors

Fluegel, T.,Schacherl, M. (deposition date: 2023-12-06, release date: 2024-03-06, Last modification date: 2024-04-24)

Primary citation

Flugel, T.,Schacherl, M.,Unbehaun, A.,Schroeer, B.,Dabrowski, M.,Burger, J.,Mielke, T.,Sprink, T.,Diebolder, C.A.,Guillen Schlippe, Y.V.,Spahn, C.M.T.
Transient disome complex formation in native polysomes during ongoing protein synthesis captured by cryo-EM.
Nat Commun, 15:1756-1756, 2024

PubMed Abstract: Structural studies of translating ribosomes traditionally rely on in vitro assembly and stalling of ribosomes in defined states. To comprehensively visualize bacterial translation, we reactivated ex vivo-derived E. coli polysomes in the PURE in vitro translation system and analyzed the actively elongating polysomes by cryo-EM. We find that 31% of 70S ribosomes assemble into disome complexes that represent eight distinct functional states including decoding and termination intermediates, and a pre-nucleophilic attack state. The functional diversity of disome complexes together with RNase digest experiments suggests that paused disome complexes transiently form during ongoing elongation. Structural analysis revealed five disome interfaces between leading and queueing ribosomes that undergo rearrangements as the leading ribosome traverses through the elongation cycle. Our findings reveal at the molecular level how bL9's CTD obstructs the factor binding site of queueing ribosomes to thwart harmful collisions and illustrate how translation dynamics reshape inter-ribosomal contacts.

PubMed: 38409277
DOI: 10.1038/s41467-024-46092-3

Experimental method

ELECTRON MICROSCOPY (3.59 Å)