The hybrid state of tRNA binding is an authentic translation elongation intermediate

• Published in: Nature structural & molecular biology Vol. 13; no. 3; pp. 234 - 241
• Main Authors: Dorner, Silke, Brunelle, Julie L, Sharma, Divya, Green, Rachel
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.03.2006; Nature Publishing Group
• Subjects: Binding sites; Biochemistry; Biological Microscopy; Biomedical and Life Sciences; Escherichia coli - genetics; Escherichia coli - metabolism; Genetic translation; Kinetics; Life Sciences; Membrane Biology; Molecular biology; Mutation - genetics; Peptide Chain Elongation, Translational - drug effects; Peptide Elongation Factor G - metabolism; Physiological aspects; Protein binding; Protein Structure; Ribonucleic acid; Ribosomes - genetics; Ribosomes - metabolism; RNA; RNA, Ribosomal - genetics; RNA, Ribosomal - metabolism; RNA, Transfer - genetics; RNA, Transfer - metabolism; RNA, Transfer, Met - genetics; RNA, Transfer, Met - metabolism; Sparsomycin - pharmacology; Transfer RNA; Translocation; United States
• ISSN: 1545-9993; 1545-9985
• DOI: 10.1038/nsmb1060

The GTPase elongation factor (EF)-G is responsible for promoting the translocation of the messenger RNA–transfer RNA complex on the ribosome, thus opening up the A site for the next aminoacyl-tRNA. Chemical modification and cryo-EM studies have indicated that tRNAs can bind the ribosome in an alternative 'hybrid' state after peptidyl transfer and before translocation, though the relevance of this state during translation elongation has been a subject of debate. Here, using pre–steady-state kinetic approaches and mutant analysis, we show that translocation by EF-G is most efficient when tRNAs are bound in a hybrid state, supporting the argument that this state is an authentic intermediate during translation.