Identification of thermodynamically relevant interactions between EF-Tu and backbone elements of tRNA

Jeffrey A. Pleiss, Olke C. Uhlenbeck*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

38 Scopus citations


A set of 45 different tRNAs, each containing a single deoxynucleotide substitution covering the upper half of the molecule was used in conjunction with a high-throughput ribonuclease protection assay to investigate the thermodynamic role of 2′ hydroxyl groups in stabilizing a complex with elongation factor Tu (EF-Tu) from Thermus thermophilus. Five distinct 2′ hydroxyl groups were identified where substitution with a proton resulted in an approximately tenfold decrease in the binding affinity. The same five 2′ hydroxyl groups reduced the affinity of the interaction with the nearly identical Thermus aquaticus EF-Tu. Four of these 2′ hydroxyl groups were observed to form hydrogen bonds in a co-crystal structure of tRNAPhe and T. aquaticus EF-Tu, while the fifth 2′ hydroxyl group can be associated with an intramolecular hydrogen bond in the tRNA. However, four additional hydrogen bonds to 2′ hydroxyl groups observed in the crystal structure show no thermodynamic effect upon disruption. Some of these discrepancies may be reconciled based on the unbound structures of the protein and RNA.

Original languageEnglish (US)
Pages (from-to)895-905
Number of pages11
JournalJournal of Molecular Biology
Issue number5
StatePublished - May 18 2001


  • Conformational rearrangements
  • Deoxynucleotide substitutions
  • Elongation factor Tu (EF-Tu)
  • Thermodynamic footprinting
  • tRNA

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology


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