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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 May 1;88(9):3872–3876. doi: 10.1073/pnas.88.9.3872

Anticodon-dependent aminoacylation of a noncognate tRNA with isoleucine, valine, and phenylalanine in vivo.

L Pallanck 1, L H Schulman 1
PMCID: PMC51555  PMID: 2023934

Abstract

An assay based on the initiation of protein synthesis in Escherichia coli has been used to explore the role of the anticodon in tRNA identity in vivo. Mutations were introduced into the initiator tRNA to change the wild-type anticodon from CAU (methionine) to GAU (isoleucine), GAC (valine), and GAA (phenylalanine), where each derivative differs from the preceding by a single base change in the anticodon (underlined). These changes were sufficient to cause the mutant tRNAs to be aminoacylated by the corresponding aminoacyl-tRNA synthetases based on the amino acid inserted into protein from complementary initiation codons. Construction of additional single base anticodon variants (GUU, GGU, GCC, GUC, GCA, and UAA) showed that all three anticodon bases specify isoleucine and phenylalanine identity and that both the middle and the third anticodon bases are important for valine identity in vivo.

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Selected References

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

  1. Ahrweiler P. M., Frieden C. Construction of a fol mutant strain of Escherichia coli for use in dihydrofolate reductase mutagenesis experiments. J Bacteriol. 1988 Jul;170(7):3301–3304. doi: 10.1128/jb.170.7.3301-3304.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amann E., Brosius J., Ptashne M. Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli. Gene. 1983 Nov;25(2-3):167–178. doi: 10.1016/0378-1119(83)90222-6. [DOI] [PubMed] [Google Scholar]
  3. Baccanari D. P., Stone D., Kuyper L. Effect of a single amino acid substitution on Escherichia coli dihydrofolate reductase catalysis and ligand binding. J Biol Chem. 1981 Feb 25;256(4):1738–1747. [PubMed] [Google Scholar]
  4. Bare L., Uhlenbeck O. C. Aminoacylation of anticodon loop substituted yeast tyrosine transfer RNA. Biochemistry. 1985 Apr 23;24(9):2354–2360. doi: 10.1021/bi00330a034. [DOI] [PubMed] [Google Scholar]
  5. Ben-Bassat A., Bauer K., Chang S. Y., Myambo K., Boosman A., Chang S. Processing of the initiation methionine from proteins: properties of the Escherichia coli methionine aminopeptidase and its gene structure. J Bacteriol. 1987 Feb;169(2):751–757. doi: 10.1128/jb.169.2.751-757.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  7. Bruce A. G., Uhlenbeck O. C. Specific interaction of anticodon loop residues with yeast phenylalanyl-tRNA synthetase. Biochemistry. 1982 Aug 17;21(17):3921–3926. doi: 10.1021/bi00260a003. [DOI] [PubMed] [Google Scholar]
  8. Chattapadhyay R., Pelka H., Schulman L. H. Initiation of in vivo protein synthesis with non-methionine amino acids. Biochemistry. 1990 May 8;29(18):4263–4268. doi: 10.1021/bi00470a001. [DOI] [PubMed] [Google Scholar]
  9. Eilers M., Schatz G. Binding of a specific ligand inhibits import of a purified precursor protein into mitochondria. Nature. 1986 Jul 17;322(6076):228–232. doi: 10.1038/322228a0. [DOI] [PubMed] [Google Scholar]
  10. Hirel P. H., Schmitter M. J., Dessen P., Fayat G., Blanquet S. Extent of N-terminal methionine excision from Escherichia coli proteins is governed by the side-chain length of the penultimate amino acid. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8247–8251. doi: 10.1073/pnas.86.21.8247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kleina L. G., Masson J. M., Normanly J., Abelson J., Miller J. H. Construction of Escherichia coli amber suppressor tRNA genes. II. Synthesis of additional tRNA genes and improvement of suppressor efficiency. J Mol Biol. 1990 Jun 20;213(4):705–717. doi: 10.1016/S0022-2836(05)80257-8. [DOI] [PubMed] [Google Scholar]
  12. Knowlton R. G., Soll L., Yarus M. Dual specificity of su+ 7 tRNA. Evidence for translational discrimination. J Mol Biol. 1980 Jun 5;139(4):705–720. doi: 10.1016/0022-2836(80)90056-x. [DOI] [PubMed] [Google Scholar]
  13. McClain W. H., Foss K. Changing the acceptor identity of a transfer RNA by altering nucleotides in a "variable pocket". Science. 1988 Sep 30;241(4874):1804–1807. doi: 10.1126/science.2459773. [DOI] [PubMed] [Google Scholar]
  14. McClain W. H., Foss K. Changing the identity of a tRNA by introducing a G-U wobble pair near the 3' acceptor end. Science. 1988 May 6;240(4853):793–796. doi: 10.1126/science.2452483. [DOI] [PubMed] [Google Scholar]
  15. McClain W. H., Foss K. Nucleotides that contribute to the identity of Escherichia coli tRNA(Phe). J Mol Biol. 1988 Aug 20;202(4):697–709. doi: 10.1016/0022-2836(88)90551-7. [DOI] [PubMed] [Google Scholar]
  16. Muramatsu T., Nishikawa K., Nemoto F., Kuchino Y., Nishimura S., Miyazawa T., Yokoyama S. Codon and amino-acid specificities of a transfer RNA are both converted by a single post-transcriptional modification. Nature. 1988 Nov 10;336(6195):179–181. doi: 10.1038/336179a0. [DOI] [PubMed] [Google Scholar]
  17. Normanly J., Abelson J. tRNA identity. Annu Rev Biochem. 1989;58:1029–1049. doi: 10.1146/annurev.bi.58.070189.005121. [DOI] [PubMed] [Google Scholar]
  18. Normanly J., Kleina L. G., Masson J. M., Abelson J., Miller J. H. Construction of Escherichia coli amber suppressor tRNA genes. III. Determination of tRNA specificity. J Mol Biol. 1990 Jun 20;213(4):719–726. doi: 10.1016/S0022-2836(05)80258-X. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Normanly J., Ogden R. C., Horvath S. J., Abelson J. Changing the identity of a transfer RNA. Nature. 1986 May 15;321(6067):213–219. doi: 10.1038/321213a0. [DOI] [PubMed] [Google Scholar]
  21. Prendergast N. J., Delcamp T. J., Smith P. L., Freisheim J. H. Expression and site-directed mutagenesis of human dihydrofolate reductase. Biochemistry. 1988 May 17;27(10):3664–3671. doi: 10.1021/bi00410a022. [DOI] [PubMed] [Google Scholar]
  22. Sampson J. R., DiRenzo A. B., Behlen L. S., Uhlenbeck O. C. Nucleotides in yeast tRNAPhe required for the specific recognition by its cognate synthetase. Science. 1989 Mar 10;243(4896):1363–1366. doi: 10.1126/science.2646717. [DOI] [PubMed] [Google Scholar]
  23. Schulman L. H., Goddard J. P. Loss of methionine acceptor activity resulting from a base change in the anticodon of Escherichia coli formylmethionine transfer ribonucleic acid. J Biol Chem. 1973 Feb 25;248(4):1341–1345. [PubMed] [Google Scholar]
  24. Schulman L. H., Pelka H. Anticodon switching changes the identity of methionine and valine transfer RNAs. Science. 1988 Nov 4;242(4879):765–768. doi: 10.1126/science.3055296. [DOI] [PubMed] [Google Scholar]
  25. Schulman L. H. Recognition of tRNAs by aminoacyl-tRNA synthetases. Prog Nucleic Acid Res Mol Biol. 1991;41:23–87. [PubMed] [Google Scholar]
  26. Seong B. L., RajBhandary U. L. Escherichia coli formylmethionine tRNA: mutations in GGGCCC sequence conserved in anticodon stem of initiator tRNAs affect initiation of protein synthesis and conformation of anticodon loop. Proc Natl Acad Sci U S A. 1987 Jan;84(2):334–338. doi: 10.1073/pnas.84.2.334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Seong B. L., RajBhandary U. L. Mutants of Escherichia coli formylmethionine tRNA: a single base change enables initiator tRNA to act as an elongator in vitro. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8859–8863. doi: 10.1073/pnas.84.24.8859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Smith D. R., Calvo J. M. Nucleotide sequence of the E coli gene coding for dihydrofolate reductase. Nucleic Acids Res. 1980 May 24;8(10):2255–2274. doi: 10.1093/nar/8.10.2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Varshney U., RajBhandary U. L. Initiation of protein synthesis from a termination codon. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1586–1590. doi: 10.1073/pnas.87.4.1586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Young R. A. Transcription termination in the Escherichia coli ribosomal RNA operon rrnC. J Biol Chem. 1979 Dec 25;254(24):12725–12731. [PubMed] [Google Scholar]

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