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Idiosyncratic tuning of tRNAs to achieve uniform ribosome binding

Nature Structural & Molecular Biology volume 12, pages 788–793 (2005)Cite this article

Abstract

The binding of seven tRNA anticodons to their complementary codons on Escherichia coli ribosomes was substantially impaired, as compared with the binding of their natural tRNAs, when they were transplanted into tRNA2Ala. An analysis of chimeras composed of tRNA2Ala and various amounts of either tRNA3Gly or tRNA2Arg indicates that the presence of the parental 32-38 nucleotide pair is sufficient to restore ribosome binding of the transplanted anticodons. Furthermore, mutagenesis of tRNA2Ala showed that its highly conserved A32-U38 pair serves to weaken ribosome affinity. We propose that this negative binding determinant is used to offset the very tight codon-anticodon interaction of tRNA2Ala. This suggests that each tRNA sequence has coevolved with its anticodon to tune ribosome affinity to a value that is the same for all tRNAs.

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Figure 1: The ACG anticodon transplanted into tRNA2Ala binds poorly to E. coli 70S ribosomes.
Figure 2: Chimeric tRNAs combining tRNA2Ala (black) with tRNA3Gly (red, upper row) or tRNA2Arg (green, lower row).
Figure 3: Rate of dissociation of tRNA2Ala with different 32-38 pairs from the A site.
Figure 4: Ribosome binding determinants in the tRNA2Ala body.

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Acknowledgements

This work was supported by US National Institutes of Health Grant GM 37552 to O.C.U. We would also like to thank M. Saks for critically reading the manuscript.

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Author notes

  1. Richard P Fahlman

    Present address: Centre for Blood Research, University of British Columbia, Vancouver, British Columbia, V6T 1Z3, Canada

Authors and Affiliations

  1. Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, 2145 Sheridan Road, Evanston, 60208, Illinois, USA

    Mikołaj Olejniczak, Taraka Dale, Richard P Fahlman & Olke C Uhlenbeck

  2. Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, Poznań, 61-704, Poland

    Mikołaj Olejniczak

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Correspondence to Olke C Uhlenbeck.

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Olejniczak, M., Dale, T., Fahlman, R. et al. Idiosyncratic tuning of tRNAs to achieve uniform ribosome binding. Nat Struct Mol Biol 12, 788–793 (2005). https://doi.org/10.1038/nsmb978

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