Dimers of <i>Thermus thermophilus</i> elongation factor Ts are required for its function as a nucleotide exchange factor of elongation factor Tu

Nesper, Martina; Nock, Steffen; Sedlák, Erik; Antalı́k, Marián; Podhradský, Dušan; Sprinzl, Mathias

doi:10.1046/j.1432-1327.1998.2550081.x

Cited by 14 publications

(11 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Crystal structures have revealed that the complex of Tt Tu/Ts is a heterotetramer, whereas the complex of Ec Tu/Ts is a heterodimer (29,30). Because Tt Ts dimer formation is required for its function as a nucleotide exchange factor (31), it would be expected that the monomeric Ec Ts would not function effectively with Tt Tu. However, based on the crystal structures, Tt Tu and Tt Ts interact through almost the same bipartite interface as that between Ec Tu and Ec Ts (29,30), and based on in vivo studies, Tt Tu has been suggested to participate in protein synthesis in E. coli (32).…”

Section: Discussionmentioning

confidence: 99%

Reconstitution of translation from Thermus thermophilus reveals a minimal set of components sufficient for protein synthesis at high temperatures and functional conservation of modern and ancient translation components

Zhou

Asahara

Gaucher

et al. 2012

Nucleic Acids Research

View full text Add to dashboard Cite

Thermus thermophilus is a thermophilic model organism distantly related to the mesophilic model organism E. coli . We reconstituted protein translation of Thermus thermophilus in vitro from purified ribosomes, transfer ribonucleic acids (tRNAs) and 33 recombinant proteins. This reconstituted system was fully functional, capable of translating natural messenger RNA (mRNA) into active full-length proteins at temperatures up to 65°C and with yields up to 60 μg/ml. Surprisingly, the synthesis of active proteins also occurred at 37°C, a temperature well below the minimal growth temperature for T. thermophilus . A polyamine was required, with tetraamine being most effective, for translation at both high and low temperatures. Using such a defined in vitro system, we demonstrated a minimal set of components that are sufficient for synthesizing active proteins at high temperatures, the functional compatibility of key translation components between T. thermophilus and E. coli , and the functional conservation of a number of resurrected ancient elongation factors. This work sets the stage for future experiments that apply abundant structural information to biochemical characterization of protein translation and folding in T. thermophilus . Because it contains significantly reduced nucleases and proteases, this reconstituted thermostable cell-free protein synthesis system may enable in vitro engineering of proteins with improved thermostability.

show abstract

Section: Discussionmentioning

confidence: 99%

Reconstitution of translation from Thermus thermophilus reveals a minimal set of components sufficient for protein synthesis at high temperatures and functional conservation of modern and ancient translation components

Zhou

Asahara

Gaucher

et al. 2012

Nucleic Acids Research

View full text Add to dashboard Cite

show abstract

“…Bacterial EF-Ts and eukaryotic EF-1 b, an analog of EF-Ts, are comprised of only one domain per polypeptide and have an approximate molecular weight of 30,000. It has been reported that dimerization of Thermus thermophilus EF-Ts is required for its nucleotide exchange function, in contrast with E. coli EF-Ts, which functions as monomers (Nesper et al, 1998). Perhaps chloroplast EF-Ts proteins also require dimerization for their function and that could be accomplished by having two EF-Ts domains in the same polypeptide.…”

Section: Discussionmentioning

confidence: 99%

Chloroplast Elongation Factor Ts Pro-Protein Is an Evolutionarily Conserved Fusion with the S1 Domain-Containing Plastid-Specific Ribosomal Protein-7

2004

View full text Add to dashboard Cite

The components of chloroplast translation are similar to those of prokaryotic translation but contain some additional unique features. Proteomic analysis of the Chlamydomonas reinhardtii chloroplast ribosome identified an S1-like protein, plastid-specific ribosomal protein-7 (PSRP-7), as a stoichiometric component of the 30S subunit. Here, we report that PSRP-7 is part of a polyprotein that contains PSRP-7 on its amino end and two translation elongation factor Ts (EF-Ts) domains at the carboxy end. We named this polyprotein PETs (for polyprotein of EF-Ts). Pets is a single-copy gene containing the only chloroplast PSRP-7 and EF-Ts sequences found in the C. reinhardtii genome. The pets precursor transcript undergoes alternative splicing to generate three mRNAs with open reading frames (ORFs) of 1.68, 1.8, and 3 kb. A 110-kD pro-protein is translated from the 3-kb ORF, and the majority of this protein is likely posttranslationally processed into the 65-kD protein PSRP-7 and a 55-kD EF-Ts. PETs homologs are found in Arabidopsis thaliana and rice (Oryza sativa). The conservation of the 110-kD PETs polyprotein in the plant kingdom suggests that PSRP-7 and EF-Ts function together in some aspects of chloroplast translation and that the PETs pro-protein may have a novel function as a whole

show abstract

“…SDM experiments showed than the hydrophobic interactions present in the Thermus enzyme made the dimer more resistant to dissociation (180, 250) (see "Hydrophobic Interactions" above). Mutagenesis of a hydrophobic core at the dimer interface in the T. thermophilus elongation factor EF-Ts showed that this hydrophobic core contributed to the enzyme dimerization and that dimer formation considerably contributed to the thermodynamic stability of T. thermophilus EF-Ts, (259). Methanopyrus kandleri formylmethanofuran:tetrahydromethanopterin (H 4 MPT) formyltransferase (MkFT) is monomeric and inactive at low salt concentrations.…”

Section: Intersubunit Interactions and Oligomerizationmentioning

confidence: 99%

Hyperthermophilic Enzymes: Sources, Uses, and Molecular Mechanisms for Thermostability

Vieille¹,

Zeikus

2001

Microbiol Mol Biol Rev

1,872

1,476

View full text Add to dashboard Cite

Enzymes synthesized by hyperthermophiles (bacteria and archaea with optimal growth temperatures of >80°C), also called hyperthermophilic enzymes, are typically thermostable (i.e., resistant to irreversible inactivation at high temperatures) and are optimally active at high temperatures. These enzymes share the same catalytic mechanisms with their mesophilic counterparts. When cloned and expressed in mesophilic hosts, hyperthermophilic enzymes usually retain their thermal properties, indicating that these properties are genetically encoded. Sequence alignments, amino acid content comparisons, crystal structure comparisons, and mutagenesis experiments indicate that hyperthermophilic enzymes are, indeed, very similar to their mesophilic homologues. No single mechanism is responsible for the remarkable stability of hyperthermophilic enzymes. Increased thermostability must be found, instead, in a small number of highly specific alterations that often do not obey any obvious traffic rules. After briefly discussing the diversity of hyperthermophilic organisms, this review concentrates on the remarkable thermostability of their enzymes. The biochemical and molecular properties of hyperthermophilic enzymes are described. Mechanisms responsible for protein inactivation are reviewed. The molecular mechanisms involved in protein thermostabilization are discussed, including ion pairs, hydrogen bonds, hydrophobic interactions, disulfide bridges, packing, decrease of the entropy of unfolding, and intersubunit interactions. Finally, current uses and potential applications of thermophilic and hyperthermophilic enzymes as research reagents and as catalysts for industrial processes are described

show abstract

Dimers of Thermus thermophilus elongation factor Ts are required for its function as a nucleotide exchange factor of elongation factor Tu

Cited by 14 publications

References 5 publications

Reconstitution of translation from Thermus thermophilus reveals a minimal set of components sufficient for protein synthesis at high temperatures and functional conservation of modern and ancient translation components

Reconstitution of translation from Thermus thermophilus reveals a minimal set of components sufficient for protein synthesis at high temperatures and functional conservation of modern and ancient translation components

Chloroplast Elongation Factor Ts Pro-Protein Is an Evolutionarily Conserved Fusion with the S1 Domain-Containing Plastid-Specific Ribosomal Protein-7

Hyperthermophilic Enzymes: Sources, Uses, and Molecular Mechanisms for Thermostability

Contact Info

Product

Resources

About