Activation of GTP hydrolysis in mRNA-tRNA translocation by elongation factor G

Li, Wen; Liu, Zheng; Koripella, Ravi Kiran; Langlois, Robert; Sanyal, Suparna; Frank, Joachim

doi:10.1126/sciadv.1500169

Cited by 63 publications

(72 citation statements)

References 43 publications

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“…And during the reverse translocation eEF2 binds to the POST complex, which has a conformation of unrotated ribosomal subunits because no tRNAs with hybrid acceptor ends are present therein (5,38). Earlier it has been shown by kinetic (39) and structural (40) data that EF-G prefers to bind a ribosome in the state of rotated subunits and exhibits a 7-fold higher dwell time of binding for this conformation (39). It means that EF-G better interacts with PRE than POST complexes that correlates with our results (Fig.…”

Section: Condition Requirements For Reverse Translocationmentioning

confidence: 99%

Eukaryotic translation elongation factor 2 (eEF2) catalyzes reverse translocation of the eukaryotic ribosome

Susorov

Zakharov

Shuvalova

et al. 2018

Journal of Biological Chemistry

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During protein synthesis, a ribosome moves along the mRNA template and, using aminoacyl-tRNAs, decodes the template nucleotide triplets to assemble a protein amino acid sequence. This movement is accompanied by shifting of mRNA-tRNA complexes within the ribosome in a process called translocation. In living cells, this process proceeds in a unidirectional manner, bringing the ribosome to the 3′-end of mRNA, and is catalyzed by the GTPase translation elongation factor 2 (EF-G in prokaryotes, eEF2 in eukaryotes). Interestingly, the possibility of spontaneous backward translocation has been shown in vitro for bacterial ribosomes, suggesting a potential reversibility of this reaction. However, this possibility has not yet been tested in eukaryotic cells. Here, using a reconstituted mammalian translation system, we show that eukaryotic elongation factor eEF2 catalyzes ribosomal reverse translocation at one mRNA triplet. We found that this process requires a cognate tRNA in the ribosomal E-site and cannot occur spontaneously without eEF2. The efficiency of this reaction depended on the concentrations of eEF2 and cognate tRNAs and increased in the presence of nonhydrolyzable GTP analogues. Of note, ADP-ribosylation of eEF2 domain IV blocked reverse translocation, suggesting a crucial role of interactions of this domain with the ribosome for the catalysis of the reaction. In summary, our findings indicate that eEF2 is able to induce ribosomal translocation in forward and backward directions highlighting the universal mechanism of tRNA-mRNA movements within the ribosome.

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Section: Condition Requirements For Reverse Translocationmentioning

confidence: 99%

Eukaryotic translation elongation factor 2 (eEF2) catalyzes reverse translocation of the eukaryotic ribosome

Susorov

Zakharov

Shuvalova

et al. 2018

Journal of Biological Chemistry

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“…[30][31][32][33][34][35] However, our results indicate that performing such long minimizations may lead to thermodynamic destabilization of proteins which means that these protein structures may diverge from the near-native attraction basin. As a result, using an unphysical protein structure in MD is perhaps not a problem only if a full conformational space is sampled; however, such ergodicity requirement is rarely met in practice.…”

Section: A Energy Decompositionmentioning

confidence: 84%

Communication: Entropic measure to prevent energy over-minimization in molecular dynamics simulations

Rydzewski

Jakubowski

Nowak

2015

The Journal of Chemical Physics

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This work examines the impact of energy over-minimization on an ensemble of biological molecules subjected to the potential energy minimization procedure in vacuum. In the studied structures, long potential energy minimization stage leads to an increase of the main-and side-chain entropies in proteins. We show that such over-minimization may diverge the protein structures from the near-native attraction basin which possesses a minimum of free energy. We propose a measure based on the Pareto front of total entropy for quality assessment of minimized protein conformation. This measure may help in selection of adequate number of energy minimization steps in protein modelling and, thus, in preservation of the near-native protein conformation. C 2015 AIP Publishing LLC. [http://dx

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“…part of the elongation cycle is visualized in asample containing amutant of EF-G (Figure 23 right). [83] Literally,cryo-EM is now able to tell us astory from asingle sample of molecules in equilibrium, showing how the molecule changes its shape and binds or sheds ligands. This potential for resolving dynamic changes of molecules in equilibrium is augmented by the development of timeresolved techniques that are able to trap short-lived states evolving in an on-equilibrium experiment.…”

Section: Move To Columbia University-story In Asamplementioning

confidence: 99%

Single‐Particle Reconstruction of Biological Molecules—Story in a Sample (Nobel Lecture)

Frank

2018

Angew Chem Int Ed

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Activation of GTP hydrolysis in mRNA-tRNA translocation by elongation factor G

Cited by 63 publications

References 43 publications

Eukaryotic translation elongation factor 2 (eEF2) catalyzes reverse translocation of the eukaryotic ribosome

Eukaryotic translation elongation factor 2 (eEF2) catalyzes reverse translocation of the eukaryotic ribosome

Communication: Entropic measure to prevent energy over-minimization in molecular dynamics simulations

Single‐Particle Reconstruction of Biological Molecules—Story in a Sample (Nobel Lecture)

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