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. 1989 Jun;86(12):4397–4401. doi: 10.1073/pnas.86.12.4397

tRNA-tRNA interactions within cellular ribosomes.

D Smith 1, M Yarus 1
PMCID: PMC287276  PMID: 2499882

Abstract

We describe an assay that converts the effects of tRNA-tRNA contacts at two particular codons into a quantitative effect on beta-galactosidase level. The assay measures the separate and combined efficiency of suppression at adjacent nonsense codons in vivo using a set of specially created homologous messages. In a survey of distal anticodon arm substitutions, we find that particular mutant tRNAs occupying the P-site reduce the apparent efficiency of the suppressor tRNA reading the A-site codon by factors of 2-170. By using measured tRNA-tRNA distances and the crystallographic tRNA structure, we propose a model of the tRNA-tRNA-mRNA complex. In the model, the anticodon loops of the P-site and A-site tRNAs contact one another in a way that is consistent with our combined tRNA efficiency data. These results suggest that tRNA-tRNA interactions that modulate tRNA action are an inevitable feature of translation.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bossi L. Context effects: translation of UAG codon by suppressor tRNA is affected by the sequence following UAG in the message. J Mol Biol. 1983 Feb 15;164(1):73–87. doi: 10.1016/0022-2836(83)90088-8. [DOI] [PubMed] [Google Scholar]
  2. Dock-Bregeon A. C., Chevrier B., Podjarny A., Moras D., deBear J. S., Gough G. R., Gilham P. T., Johnson J. E. High resolution structure of the RNA duplex [U(U-A)6A]2. Nature. 1988 Sep 22;335(6188):375–378. doi: 10.1038/335375a0. [DOI] [PubMed] [Google Scholar]
  3. Fairclough R. H., Cantor C. R. The distance between the anticodon loops of two tRNAs bound to the 70 S Escherichia coli ribosome. J Mol Biol. 1979 Aug 25;132(4):575–586. doi: 10.1016/0022-2836(79)90375-9. [DOI] [PubMed] [Google Scholar]
  4. Holbrook S. R., Sussman J. L., Warrant R. W., Kim S. H. Crystal structure of yeast phenylalanine transfer RNA. II. Structural features and functional implications. J Mol Biol. 1978 Aug 25;123(4):631–660. doi: 10.1016/0022-2836(78)90210-3. [DOI] [PubMed] [Google Scholar]
  5. Johnson A. E., Adkins H. J., Matthews E. A., Cantor C. R. Distance moved by transfer RNA during translocation from the A site to the P site on the ribosome. J Mol Biol. 1982 Mar 25;156(1):113–140. doi: 10.1016/0022-2836(82)90462-4. [DOI] [PubMed] [Google Scholar]
  6. Lacatena R. M., Cesareni G. Interaction between RNA1 and the primer precursor in the regulation of Co1E1 replication. J Mol Biol. 1983 Nov 5;170(3):635–650. doi: 10.1016/s0022-2836(83)80125-9. [DOI] [PubMed] [Google Scholar]
  7. McDonald J. J., Rein R. A stereochemical model of the transpeptidation complex. J Biomol Struct Dyn. 1987 Apr;4(5):729–744. doi: 10.1080/07391102.1987.10507675. [DOI] [PubMed] [Google Scholar]
  8. Miller J. H., Albertini A. M. Effects of surrounding sequence on the suppression of nonsense codons. J Mol Biol. 1983 Feb 15;164(1):59–71. doi: 10.1016/0022-2836(83)90087-6. [DOI] [PubMed] [Google Scholar]
  9. Murgola E. J., Pagel F. T., Hijazi K. A. Codon context effects in missense suppression. J Mol Biol. 1984 May 5;175(1):19–27. doi: 10.1016/0022-2836(84)90442-x. [DOI] [PubMed] [Google Scholar]
  10. Normanly J., Masson J. M., Kleina L. G., Abelson J., Miller J. H. Construction of two Escherichia coli amber suppressor genes: tRNAPheCUA and tRNACysCUA. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6548–6552. doi: 10.1073/pnas.83.17.6548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Paulsen H., Robertson J. M., Wintermeyer W. Topological arrangement of two transfer RNAs on the ribosome. Fluorescence energy transfer measurements between A and P site-bound tRNAphe. J Mol Biol. 1983 Jun 25;167(2):411–426. doi: 10.1016/s0022-2836(83)80342-8. [DOI] [PubMed] [Google Scholar]
  12. Smith D., Yarus M. Transfer RNA structure and coding specificity. I. Evidence that a D-arm mutation reduces tRNA dissociation from the ribosome. J Mol Biol. 1989 Apr 5;206(3):489–501. doi: 10.1016/0022-2836(89)90496-8. [DOI] [PubMed] [Google Scholar]
  13. Vacher J., Springer M., Buckingham R. H. Functional mutants of phenylalanine transfer RNA from Escherichia coli. EMBO J. 1985 Feb;4(2):509–513. doi: 10.1002/j.1460-2075.1985.tb03657.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Woese C. Molecular mechanics of translation: a reciprocating ratchet mechanism. Nature. 1970 May 30;226(5248):817–820. doi: 10.1038/226817a0. [DOI] [PubMed] [Google Scholar]
  15. Yarus M., Cline S. W., Wier P., Breeden L., Thompson R. C. Actions of the anticodon arm in translation on the phenotypes of RNA mutants. J Mol Biol. 1986 Nov 20;192(2):235–255. doi: 10.1016/0022-2836(86)90362-1. [DOI] [PubMed] [Google Scholar]
  16. Yarus M., Folley L. S. Sense codons are found in specific contexts. J Mol Biol. 1985 Apr 20;182(4):529–540. doi: 10.1016/0022-2836(85)90239-6. [DOI] [PubMed] [Google Scholar]
  17. Yarus M. Translational efficiency of transfer RNA's: uses of an extended anticodon. Science. 1982 Nov 12;218(4573):646–652. doi: 10.1126/science.6753149. [DOI] [PubMed] [Google Scholar]

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