Effect of streptomycin on the stoichiometry of GTP hydrolysis in a poly(U)-dependent cell-free translation system

• Published in: FEBS letters Vol. 192; no. 1; pp. 165 - 169
• Main Authors: Smailov, S.K., Gavrilova, L.P.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 11-11-1985; Elsevier
• Subjects: Biological and medical sciences; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Guanosine Triphosphate - metabolism; Hydrolysis; Kinetics; Magnesium - pharmacology; Misreading; Molecular and cellular biology; Molecular genetics; Peptide Biosynthesis; Peptide Chain Elongation, Translational - drug effects; Peptide Elongation Factor Tu - metabolism; Peptides; Poly U - genetics; Proof-reading; Protein Biosynthesis - drug effects; Ribosomes - metabolism; RNA, Transfer, Amino Acyl - metabolism; Sepharose-bound poly (U) translation technique; Streptomycin - pharmacology; Translation. Translation factors. Protein processing; Misreading; Sepharose-bound poly (U) translation technique; Proof-reading; Hydrolysis; Error; GTP; Streptomycin; In vitro translation
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(85)80065-X

The technique of Sepharose-bound template translation has been used to estimate the stoichiometry of GTP hydrolysis during peptide elongation in the presence of streptomycin. The presence of streptomycin has been shown to have no great effect on the elongation rate and the stoichiometry of GTP hydrolysis during codonspecific peptide elongation in the poly(U)-directed translation system: the molar ratio of hydrolysed GTP to incorporated phenylalanine was about 2. At the same time streptomycin exerted a significant effect during misreading when a ribosome-bound peptide in the poly(U)-programmed system was elongated by leucine or isoleucine residues: the miselongation was stimulated and hence the ratio of hydrolysed GTP per peptide bond was strongly reduced, as compared with the excessive GTP hydrolysis which is characteristic of the misreading system in the absence of streptomycin [(1984) FEBS Lett. 178, 283-287]. The conclusion has been made that streptomycin blocks the stage of correction (‘proof-reading’) following GTP hydrolysis during EF-Tu-dependent aminoacyl-tRNA binding.