The hybrid state of tRNA binding is an authentic translation elongation intermediate.
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.
Dorner
Silke
S
Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Brunelle
Julie L
JL
Sharma
Divya
D
Green
Rachel
R
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D009154:Mutation / Q000235:genetics; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects; D020653:Peptide Elongation Factor G / Q000378:metabolism; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism*; D012358:RNA, Transfer, Met / Q000235:genetics / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism; D013033:Sparsomycin / Q000494:pharmacology
D020653:Peptide Elongation Factor G; D012335:RNA, Ribosomal; D012358:RNA, Transfer, Met; D013033:Sparsomycin; D012343:RNA, Transfer