Many of the conserved nucleotides of tRNA (Phe) are not essential for ternary complex formation and peptide elongation
Nazarenko, I.A.; Harrington, K.M.; Uhlenbeck, O.C.
EMBO Journal 13(10): 2464-2471
1994
ISSN/ISBN: 0261-4189 PMID: 8194535 Document Number: 435107
An RNase protection assay was used to show that the dissociation rate constants and equilibrium constants of unmodified yeast and Escherichia coli phenylalanyl-tRNA-Phes to elongation factor Tu from E. coli were very similar to each other and to their fully modified counterparts. The affinity of aminoacylated tRNA to elongation factor Tu was substantially lower when GTP analogues were used in place of GTP, emphasizing the importance of the beta-gamma phosphate linkage in the function of G-proteins. Fourteen different mutations in conserved and semi-conserved nucleotides of yeast phenylalanyl-tRNA-Phe were tested for binding to elongation factor Tu cntdot GTP and assayed for activity in the ribosomal A- and P-sites. Most of the mutations did not severely impair the function of these tRNAs in any of the assays. This suggests that the translational machinery does not form sequence-specific interactions with the conserved nucleotides of tRNA.