Rapid ribosomal translocation depends on the conserved 18-55 base pair in P-site transfer RNA

Abstract

The L shape of tRNA is stabilized by the 'tertiary core' region, which contains base-pairing interactions between the D and T loops. Distortions of the L shape accompany tRNA movement across the ribosomal surface. Here, using single-turnover rapid kinetics assays, we determine the effects of mutations within the tertiary core of P site–bound tRNAfMet on three measures of the rate of translocation, the part of the elongation cycle involving the most extensive tRNA movement. Mutations in the strictly conserved G18·U55 base pair result in as much as an 80-fold decrease in the rate of translocation, demonstrating the importance of the 18-55 interaction for rapid translocation. This implicates the core region as a locus for functionally important dynamic interactions with the ribosome and leads to the proposal that translocation of ribosome-bound tRNAs may be sequential rather than concerted.

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Figure 1: Crystal structure of E. coli tRNAfMet bound to Met-RS (PDB entry 2FMT).
Figure 2: Time course of EF-G–dependent translocation.
Figure 3: Interaction sites between the ribosome and the tertiary core region of tRNA8 (PDB entries 1GIX and 1GIY).
Figure 4

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Acknowledgements

This work was supported by US National Institutes of Health grants to B.S.C. (GM071014) and Y.M.H. (GM56662). C.M.Z. is supported by a postdoctoral fellowship from the American Heart Association, Pennsylvania-Delaware Affiliate.

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Correspondence to Barry S Cooperman.

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Pan, D., Kirillov, S., Zhang, C. et al. Rapid ribosomal translocation depends on the conserved 18-55 base pair in P-site transfer RNA. Nat Struct Mol Biol 13, 354–359 (2006). https://doi.org/10.1038/nsmb1074

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