8PKL

Escherichia coli paused disome complex (leading 70S non-rotated closed PRE state)

This is a non-PDB format compatible entry.

Summary for 8PKL

• Entry DOI: 10.2210/pdb8pkl/pdb
• EMDB information: 17743
• Descriptor: 50S ribosomal protein L36, 16S ribosomal RNA, Small ribosomal subunit protein uS2, ... (66 entities in total)
• Functional Keywords: ribosome, polysome, tranlsation, elongation, pausing, disome, collision, pure system
• Biological source: Escherichia coli; More
• Total number of polymer chains: 60
• Total formula weight: 2293943.67

Authors

Fluegel, T.,Schacherl, M. (deposition date: 2023-06-26, release date: 2024-03-06, Last modification date: 2024-10-23)

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 (3.09 Å)