Transfer RNA Dynamics in Bacterial Protein Synthesis
Studies of tRNA Movement on the Ribosome During the Elongation Cycle of Bacterial Protein Synthesis
Omschrijving
Protein biosynthesis, also called translation,refers to mRNA-programmed synthesis of a polypeptideon the ribosome. tRNAs are required to moveefficiently between several positions on the ribosomeduring the elongation phase of translation. Althoughstructures of tRNA bound to ribosomes are available,the dynamics and mechanism of tRNA movement andstructural change are incompletely understood. Theclassic L-shape of tRNA undergoes considerabledistortion during the elongation cycle, leading tothe suggestion that the tRNA tertiary core region,which stabilizes the L-shape, actively interacts withthe ribosome. Here, using fluorescence labeling oftRNA and mutagenesis of highly conserved nucleotideswithin the tertiary core, combined with binding andsingle turnover rapid kinetics assays, we monitortRNA movement on the ribosome, examine the importanceof the tertiary core region for each step of theelongation cycle, identify interactions andintermediates that are important for the accuracy andefficiency of protein biosynthesis, and determineantibiotic effects on elongation.
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