Abstract
The ribosome is a macromolecular assembly that is responsible for protein biosynthesis following genetic instructions in all organisms. It is composed of two unequal subunits: the smaller subunit binds messenger RNA and the anticodon end of transfer RNAs, and helps to decode the mRNA; and the larger subunit interacts with the amino-acid-carrying end of tRNAs and catalyses the formation of the peptide bonds. After peptide-bond formation, elongation factor G (EF-G) binds to the ribosome, triggering the translocation of peptidyl-tRNA from its aminoacyl site to the peptidyl site, and movement of mRNA by one codon1. Here we analyse three-dimensional cryo-electron microscopy maps of the Escherichia coli 70S ribosome in various functional states, and show that both EF-G binding and subsequent GTP hydrolysis lead to ratchet-like rotations of the small 30S subunit relative to the large 50S subunit. Furthermore, our finding indicates a two-step mechanism of translocation: first, relative rotation of the subunits and opening of the mRNA channel following binding of GTP to EF-G; and second, advance of the mRNA/(tRNA)2 complex in the direction of the rotation of the 30S subunit, following GTP hydrolysis.
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Acknowledgements
This work was supported by grants from the National Institutes of Health. We thank A. Heagle for preparing the illustrations and animated sequence, and I. Gabashvili and P. Penczek for help with image processing.
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41586_2000_BF35018597_MOESM1_ESM.mov
Movie: The Ribosome — a molecular ratchet. For instructions on how to play this movie, please visit http://www.quicktime.com (MOV 2534 kb)
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Frank, J., Agrawal, R. A ratchet-like inter-subunit reorganization of the ribosome during translocation . Nature 406, 318–322 (2000). https://doi.org/10.1038/35018597
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DOI: https://doi.org/10.1038/35018597
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