Direct evidence of an elongation factor-Tu/Ts·GTP·Aminoacyl-tRNA quaternary complex

J Biol Chem. 2014 Aug 22;289(34):23917-27. doi: 10.1074/jbc.M114.583385. Epub 2014 Jul 2.

Abstract

During protein synthesis, elongation factor-Tu (EF-Tu) bound to GTP chaperones the entry of aminoacyl-tRNA (aa-tRNA) into actively translating ribosomes. In so doing, EF-Tu increases the rate and fidelity of the translation mechanism. Recent evidence suggests that EF-Ts, the guanosine nucleotide exchange factor for EF-Tu, directly accelerates both the formation and dissociation of the EF-Tu-GTP-Phe-tRNA(Phe) ternary complex (Burnett, B. J., Altman, R. B., Ferrao, R., Alejo, J. L., Kaur, N., Kanji, J., and Blanchard, S. C. (2013) J. Biol. Chem. 288, 13917-13928). A central feature of this model is the existence of a quaternary complex of EF-Tu/Ts·GTP·aa-tRNA(aa). Here, through comparative investigations of phenylalanyl, methionyl, and arginyl ternary complexes, and the development of a strategy to monitor their formation and decay using fluorescence resonance energy transfer, we reveal the generality of this newly described EF-Ts function and the first direct evidence of the transient quaternary complex species. These findings suggest that EF-Ts may regulate ternary complex abundance in the cell through mechanisms that are distinct from its guanosine nucleotide exchange factor functions.

Keywords: Elongation Factor Ts; Elongation Factor Tu; G Protein; Guanine Nucleotide Exchange Factor (GEF); Guanosine Nucleotide Exchange Factor; Protein Synthesis; Ternary Complex; Translation; Translation Elongation Factor.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Escherichia coli / metabolism
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / metabolism*
  • Guanosine Triphosphate / metabolism*
  • Models, Molecular
  • Peptide Elongation Factor Tu / chemistry
  • Peptide Elongation Factor Tu / metabolism*
  • Peptide Elongation Factors / chemistry
  • Peptide Elongation Factors / metabolism*
  • RNA, Transfer, Amino Acyl / chemistry
  • RNA, Transfer, Amino Acyl / metabolism*
  • Ribosomes / metabolism

Substances

  • Escherichia coli Proteins
  • Peptide Elongation Factors
  • RNA, Transfer, Amino Acyl
  • elongation factor Ts
  • Guanosine Triphosphate
  • Peptide Elongation Factor Tu

Associated data

  • PDB/1OB2