Understanding the sequence specificity of tRNA binding to elongation factor Tu using tRNA mutagenesis. Journal Article uri icon

Overview

abstract

  • Measuring the binding affinities of 42 single-base-pair mutants in the acceptor and T Psi C stems of Saccharomyces cerevisiae tRNA Phe to Thermus thermophilus elongation factor Tu (EF-Tu) revealed that much of the specificity for tRNA occurs at the 49-65, 50-64, and 51-63 base pairs. Introducing the same mutations at the three positions into Escherichia coli tRNA CAG Leu resulted in similar changes in binding affinity. Swapping the three pairs from several E. coli tRNAs into yeast tRNA Phe resulted in chimeras with EF-Tu binding affinities similar to those for the donor tRNA. Finally, analysis of double- and triple-base-pair mutants of tRNA Phe showed that the thermodynamic contributions at the three sites are additive, permitting reasonably accurate prediction of the EF-Tu binding affinity for all E. coli tRNAs. Thus, it appears that the thermodynamic contributions of three base pairs in the T Psi C stem primarily account for tRNA binding specificity to EF-Tu.

publication date

  • March 13, 2009

has subject area

has restriction

  • green

Date in CU Experts

  • March 13, 2015 12:56 PM

Full Author List

  • Schrader JM; Chapman SJ; Uhlenbeck OC

author count

  • 3

Other Profiles

Electronic International Standard Serial Number (EISSN)

  • 1089-8638

Additional Document Info

start page

  • 1255

end page

  • 1264

volume

  • 386

issue

  • 5