Open AccessThe Complex Formation between Escherichia coli Aminoacyl‐tRNA, Elongation Factor Tu and GTP
Author(s) -
WAGNER Thomas,
SPRINZL Mathias
Publication year - 1980
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1980.tb04714.x
Subject(s) - ef tu , transfer rna , escherichia coli , gtp' , aminoacyl trna , elongation factor , elongation , gtpase , aminoacyl trna synthetase , chemistry , biology , biochemistry , rna , gene , enzyme , materials science , ribosome , ultimate tensile strength , metallurgy
The interaction between Escherichia coli aminoacyl‐tRNAs and elongation factor Tu (EF‐Tu) · GTP was examined. Ternary complex formation with Phe‐tRNA Phe and Lys‐tRNA Lys was compared to that with the respective misaminoacylated Tyr‐tRNA Phe and Phe‐tRNA Lys . There was no pronounced difference in the efficiency of aminoacyl‐tRNA · EF‐Tu · GTP complex formation between Phe‐tRNA Phe and Tyr‐tRNA Phe . However, Phe‐tRNA Lys was bound preferentially to EF‐Tu · GTP as compared to Lys‐tRNA Lys . This was shown by the ability of EF‐Tu · GTP to prevent the hydrolysis of the aminoacyl ester linkage of the aminoacyl‐tRNA species. Furthermore, gel filtration of ternary complexes revealed that the complex formed with the misaminoacylated tRNA LyS was also more stable than the one formed with the correctly aminoacylated tRNA Lys . Both misaminoacylated aminoacyl‐tRNA species could participate in the ribosomal peptide elongation reaction. Poly(U)‐directed synthesis of poly(Tyr) using Tyr‐tRNA Phe occurred to a comparable extent as the synthesis of poly(Phe) with Phe‐tRNA Phe . In the translation of poly(A) using native Lys‐tRNA Lys , poly(Lys) reached a lower level than poly(Phe) when Phe‐tRNA LyS was used. It is concluded that the side‐chain of the amino acid linked to a tRNA affects the efficiency of the aminoacyl‐tRNA · EF‐Tu · GTP ternary complex formation.