Allosteric vs. spontaneous exit-site (E-site) tRNA dissociation early in protein synthesis

作者：Chen Chunlai; Stevens Benjamin; Kaur Jaskiran; Smilansky Zeev; Cooperman Barry S^*; Goldman Yale E

来源：Proceedings of the National Academy of Sciences, 2011, 108(41): 16980-16985.

DOI：10.1073/pnas.1106999108

• FRET
• alternating wavelength laser excitation (ALEX)
• multi-translation cycles
• classical state
• hybrid state

During protein synthesis, deacylated transfer RNAs leave the ribosome via an exit (E) site after mRNA translocation. How the ribosome regulates tRNA dissociation and whether functional linkages between the aminoacyl (A) and E sites modulate the dynamics of protein synthesis have long been debated. Using single molecule fluorescence resonance energy transfer experiments, we find that, during early cycles of protein elongation, tRNAs are often held in the E site until being allosterically released when the next aminoacyl tRNA binds to the A site. This process is regulated by the length and sequence of the nascent peptide and by the conformational state, detected by tRNA proximity, prior to translocation. In later cycles, E-site tRNA dissociates spontaneously. Our results suggest that the distribution of pretranslocation tRNA states and posttranslocation pathways are correlated within each elongation cycle via communication between distant subdomains in the ribosome, but that this correlation between elongation cycle intermediates does not persist into succeeding cycles.

• 出版日期2011-10-11