Structure of eEF3 and the mechanism of transfer RNA release from the E-site

  title={Structure of eEF3 and the mechanism of transfer RNA release from the E-site},
  author={Christian Brix Folsted Andersen and Thomas Becker and Michael Blau and Monika Anand and Mario Hali{\'c} and Bharvi Balar and Thorsten Mielke and Thomas Boesen and Jan Skov Pedersen and Christian M. T. Spahn and Terri Goss Kinzy and Gregers Rom Andersen and Roland Beckmann},
Elongation factor eEF3 is an ATPase that, in addition to the two canonical factors eEF1A and eEF2, serves an essential function in the translation cycle of fungi. eEF3 is required for the binding of the aminoacyl-tRNA–eEF1A–GTP ternary complex to the ribosomal A-site and has been suggested to facilitate the clearance of deacyl-tRNA from the E-site. Here we present the crystal structure of Saccharomyces cerevisiae eEF3, showing that it consists of an amino-terminal HEAT repeat domain, followed… 
eEF3 promotes late stages of tRNA translocation on the ribosome
Cryo-EM analysis of ex vivo e EF3-ribosome complexes shows that eEF3 facilitates late steps of translocation by favoring non-rotated ribosomal states as well as by opening the L1 stalk to release the E-site tRNA.
Mutations in the Chromodomain-like Insertion of Translation Elongation Factor 3 Compromise Protein Synthesis through Reduced ATPase Activity*
Background: Eukaryotic translation elongation factor 3 (eEF3) is a ribosome binding ATPase essential for fungal protein synthesis. Results: Mutations in the chromodomain-like insertion in an ATPase
Yeast translation elongation factor eEF3 promotes late stages of tRNA translocation
It is shown that eEF3 acts at the mRNA–tRNA translocation step by promoting the dissociation of the tRNA from the E site, but independent of aminoacyl‐tRNA recruitment to the A site, enabling presentation of a revised yeast translation elongation cycle.
EttA regulates translation by binding to the ribosomal E site and restricting ribosome-tRNA dynamics
This work represents the first example, to the authors' knowledge, in which the detailed molecular mechanism of any ABC-F family protein has been determined and establishes a framework for elucidating the mechanisms of other regulatory translation factors.
The Unique Evolutionary Distribution of Eukaryotic Elongation Factor 3
Critical residues of the two ABC domains involved in nucleotide binding and hydrolysis were highly conserved in all the putative eEF3 homologs identified, supporting the functional role of the homologys as ATPases.
The role of ABCE1 in eukaryotic posttermination ribosomal recycling.
Interplay between an ATP-binding cassette F protein and the ribosome from Mycobacterium tuberculosis
Cryo-electron microscopy structures of EttA from Mycobacterium tuberculosis reveal a dynamic interplay between MtbEttA and the Mtb ribosome, providing insights into the mechanism of translational regulation by ETTA-like proteins.
Ribosome recycling step in yeast cytoplasmic protein synthesis is catalyzed by eEF3 and ATP
It is shown that a Saccharomyces cerevisiae model PoTC was disassembled by ATP and eukaryotic elongation factor 3 (eEF3), and no 40S•mRNA complex was observed, indicating that eEF3 action promotes ribosome recycling, not reinitiation.
Directed hydroxyl radical probing reveals Upf1 binding to the 80S ribosomal E site rRNA at the L1 stalk
The results shed light on the interaction between Upf1 and the ribosome, and suggest that upf1 may specifically engage a classical-state ribosomes during translation.
Ribosome profiling analysis of eEF3-depleted Saccharomyces cerevisiae
Surprisingly, depletion of eEF3 leads to a relative decrease in P-site proline stalling, which is hypothesised to be a secondary effect of generally decreased translation and/or decreased competition for the E-site with eIF5A.


Domain movements of elongation factor eEF2 and the eukaryotic 80S ribosome facilitate tRNA translocation
An 11.7‐Å‐resolution cryo‐EM map of the yeast 80S·eEF2 complex in the presence of the antibiotic sordarin was interpreted in molecular terms, revealing large conformational changes within eEF2 and
The N Terminus of Eukaryotic Translation Elongation Factor 3 Interacts with 18 S rRNA and 80 S Ribosomes*
Elongation factor-3 (EF-3) is an essential fungal-specific translation factor which exhibits a strong ribosome-dependent ATPase activity and has sequence homologies that may predict domains critical
Structure of the signal recognition particle interacting with the elongation-arrested ribosome
The model shows how the S domain of SRP contacts the large ribosomal subunit at the nascent chain exit site to bind the signal sequence, and that the Alu domain reaches into the elongation-factor-binding site of the ribosome, explaining its elongation arrest activity.
The Elongation Factor 3 Unique in Higher Fungi and Essential for Protein Biosynthesis Is an E Site Factor (*)
It is demonstrated that the first and the third tRNA binding site (A and E sites, respectively) of yeast ribosomes are reciprocally linked; if the A site is occupied the E site has lost its binding capability, and vice versa, if the E sites is occupiedThe A site has a low affinity for tRNAs.
Crystal Structure of the Ribosome at 5.5 Å Resolution
The crystal structure of the complete Thermus thermophilus 70S ribosome containing bound messenger RNA and transfer RNAs (tRNAs) at 5.5 angstrom resolution is described, suggesting coupling of the 20 to 50 angstrom movements associated with tRNA translocation with intersubunit movement.
A ratchet-like inter-subunit reorganization of the ribosome during translocation
Three-dimensional cryo-electron microscopy maps of the Escherichia coli 70S ribosome in various functional states show that both EF-G binding and subsequent GTP hydrolysis lead to ratchet-like rotations of the small 30S sub unit relative to the large 50S subunit, indicating a two-step mechanism of translocation.
Mechanism of transfer RNA maturation by CCA-adding enzyme without using an oligonucleotide template
The crystal structure of the Archaeoglobus fulgidus tRNA nucleotidyltransferase in complex with tRNA is described and a single nucleotide-binding pocket exists whose specificity for both CTP and ATP is determined by the protein side chain of Arg 224 and backbone phosphates of the tRNA, which are non-complementary to and thus exclude UTP and GTP.
Role of yeast elongation factor 3 in the elongation cycle.
Investigation of the role of the polypeptide chain elongation factor 3 (EF-3) of yeast indicates that EF-3 participates in the elongation cycle by stimulating the function of EF-1 alpha in binding