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tRNA Translocation by the Eukaryotic 80S Ribosome and the Impact of GTP Hydrolysis.

<div class='caption-title'>Density maps of the three 80S•tRNA2•eEF2•GMPPNP complexes (contour level 2.3σ). 40S rRNA (yellow), 40S proteins (gray), 60S rRNA (blue), 60S proteins (gold), eEF2 (red), ap/P or P/E tRNA (green), pe/E or E/E tRNA (orange), mRNA (pink). Landmarks: 40S head (h), beak (bk), left and right foot (lf, rf), central protuberance (CP), L1 stalk (L1).</div>
<div class='caption-title'>(A–E) Movement of the 40S. Comparisons are based on a common 60S alignment. Distance changes in the 40S subunit positions resulting from rigid body transformation are color-coded in Å units. Changes of the TI-POST-1 to TI-POST-3 relative to the POST state (PDB: 5AJ0) are shown (A, C, and E) as well as the transitions between TI-POST-1 and TI-POST-2 (B) and between TI-POST-2 and TI-POST-3 (D). Rotation angles and axes were measured in Chimera.</div>
<div class='caption-title'>40S rRNA (yellow), 60S rRNA (light blue), eEF2 (red), ap/P or P/E tRNA (green), pe/E or E/E tRNA (orange), mRNA (pink), EF-G (gray).</div>
Flis J, Holm M, Rundlet EJ, Loerke J, Hilal T, Dabrowski M, Bürger J, Mielke T, Blanchard SC, Spahn CMT, Budkevich TV
OpenAccess logo Cell Rep 25 2676-2688.e7 (2018)
Related entries: 6gz3, 6gz4

Cited: 41 times
EuropePMC logo PMID: 30517857

Abstract

Translocation moves the tRNA2⋅mRNA module directionally through the ribosome during the elongation phase of protein synthesis. Although translocation is known to entail large conformational changes within both the ribosome and tRNA substrates, the orchestrated events that ensure the speed and fidelity of this critical aspect of the protein synthesis mechanism have not been fully elucidated. Here, we present three high-resolution structures of intermediates of translocation on the mammalian ribosome where, in contrast to bacteria, ribosomal complexes containing the translocase eEF2 and the complete tRNA2⋅mRNA module are trapped by the non-hydrolyzable GTP analog GMPPNP. Consistent with the observed structures, single-molecule imaging revealed that GTP hydrolysis principally facilitates rate-limiting, final steps of translocation, which are required for factor dissociation and which are differentially regulated in bacterial and mammalian systems by the rates of deacyl-tRNA dissociation from the E site.

Articles - 6gz5 mentioned but not cited (6)

  1. tRNA Translocation by the Eukaryotic 80S Ribosome and the Impact of GTP Hydrolysis. Flis J, Holm M, Rundlet EJ, Loerke J, Hilal T, Dabrowski M, Bürger J, Mielke T, Blanchard SC, Spahn CMT, Budkevich TV. Cell Rep 25 2676-2688.e7 (2018)
  2. Protein Synthesis in the Developing Neocortex at Near-Atomic Resolution Reveals Ebp1-Mediated Neuronal Proteostasis at the 60S Tunnel Exit. Kraushar ML, Krupp F, Harnett D, Turko P, Ambrozkiewicz MC, Sprink T, Imami K, Günnigmann M, Zinnall U, Vieira-Vieira CH, Schaub T, Münster-Wandowski A, Bürger J, Borisova E, Yamamoto H, Rasin MR, Ohler U, Beule D, Mielke T, Tarabykin V, Landthaler M, Kramer G, Vida I, Selbach M, Spahn CMT. Mol Cell 81 304-322.e16 (2021)
  3. Accuracy mechanism of eukaryotic ribosome translocation. Djumagulov M, Demeshkina N, Jenner L, Rozov A, Yusupov M, Yusupova G. Nature 600 543-546 (2021)
  4. Redox-sensitive E2 Rad6 controls cellular response to oxidative stress via K63-linked ubiquitination of ribosomes. Simões V, Cizubu BK, Harley L, Zhou Y, Pajak J, Snyder NA, Bouvette J, Borgnia MJ, Arya G, Bartesaghi A, Silva GM. Cell Rep 39 110860 (2022)
  5. Visualization of translation and protein biogenesis at the ER membrane. Gemmer M, Chaillet ML, van Loenhout J, Cuevas Arenas R, Vismpas D, Gröllers-Mulderij M, Koh FA, Albanese P, Scheltema RA, Howes SC, Kotecha A, Fedry J, Förster F. Nature 614 160-167 (2023)
  6. [Formula: see text]: ComplexOme-Structural Network Interpreter used to study spatial enrichment in metazoan ribosomes. Martinez-Seidel F, Hsieh YC, Walther D, Kopka J, Pereira Firmino AA. BMC Bioinformatics 22 605 (2021)


Reviews citing this publication (4)

  1. Approaches for measuring the dynamics of RNA-protein interactions. Licatalosi DD, Ye X, Jankowsky E. Wiley Interdiscip Rev RNA 11 e1565 (2020)
  2. The Structural Dynamics of Translation. Korostelev AA. Annu Rev Biochem 91 245-267 (2022)
  3. The space between notes: emerging roles for translationally silent ribosomes. Smith PR, Pandit SC, Loerch S, Campbell ZT. Trends Biochem Sci 47 477-491 (2022)
  4. Ribosome as a Translocase and Helicase. Bao C, Ermolenko DN. Biochemistry (Mosc) 86 992-1002 (2021)

Articles citing this publication (31)



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