title: Elongation factor Ts directly facilitates the formation and disassembly of the Escherichia coli elongation factor Tu·GTP·aminoacyl-tRNA ternary complex.
abstract: BACKGROUND
  Aminoacyl-tRNA (aa-tRNA) enters the ribosome in a ternary complex with the G-protein elongation factor Tu (EF-Tu) and GTP.
  
  
  RESULTS
  EF-Tu·GTP·aa-tRNA ternary complex formation and decay rates are accelerated in the presence of the nucleotide exchange factor elongation factor Ts (EF-Ts).
  
  
  CONCLUSIONS
  EF-Ts directly facilitates the formation and disassociation of ternary complex.
  
  
  CONCLUSIONS
  This system demonstrates a novel function of EF-Ts. Aminoacyl-tRNA enters the translating ribosome in a ternary complex with elongation factor Tu (EF-Tu) and GTP. Here, we describe bulk steady state and pre-steady state fluorescence methods that enabled us to quantitatively explore the kinetic features of Escherichia coli ternary complex formation and decay. The data obtained suggest that both processes are controlled by a nucleotide-dependent, rate-determining conformational change in EF-Tu. Unexpectedly, we found that this conformational change is accelerated by elongation factor Ts (EF-Ts), the guanosine nucleotide exchange factor for EF-Tu. Notably, EF-Ts attenuates the affinity of EF-Tu for GTP and destabilizes ternary complex in the presence of non-hydrolyzable GTP analogs. These results suggest that EF-Ts serves an unanticipated role in the cell of actively regulating the abundance and stability of ternary complex in a manner that contributes to rapid and faithful protein synthesis.
authors.affiliation: Departments of Physiology and Biophysics, Weill Cornell Medical College, New York, New York 10065, USA.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Burnett
authors.forename: Roger B
authors.initials: RB
authors.lastname: Altman
authors.forename: Ryan
authors.initials: R
authors.lastname: Ferrao
authors.forename: Jose L
authors.initials: JL
authors.lastname: Alejo
authors.forename: Navdep
authors.initials: N
authors.lastname: Kaur
authors.forename: Joshua
authors.initials: J
authors.lastname: Kanji
authors.forename: Scott C
authors.initials: SC
authors.lastname: Blanchard
mesh_terms: D001665:Binding Sites; D004926:Escherichia coli; D029968:Escherichia coli Proteins / Q000737:chemistry*; D006160:Guanosine Triphosphate / Q000737:chemistry; D007700:Kinetics; D008958:Models, Molecular; D010444:Peptide Elongation Factor Tu / Q000737:chemistry*; D010445:Peptide Elongation Factors / Q000737:chemistry*; D011485:Protein Binding; D055503:Protein Multimerization; D055550:Protein Stability; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: D029968:Escherichia coli Proteins; D010445:Peptide Elongation Factors; D012346:RNA, Transfer, Amino Acyl; C023512:elongation factor Ts; D006160:Guanosine Triphosphate; D010444:Peptide Elongation Factor Tu
keywords: Elongation Factor Ts; Elongation Factor Tu; G-proteins; Guanine Nucleotide Exchange Factor (GEF); Protein Synthesis; Ternary Complex; Transfer RNA (tRNA); Translation Regulation
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