Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Functional interactions in vivo between suppressor tRNA and mutationally altered ribosomal protein S4


Ribosomal mutants (rpsD) which are associated with a generally increased translational ambiguity were investigated for their effects in vivo on individual tRNA species using suppressor tRNAs as models. It was found that nonsense suppression is either increased, unaffected or decreased depending on the codon context and the rpsD allele involved as well as the nature of the suppressor tRNA. Missense suppression of AGA and AGG by glyT(SuAGA/G) tRNA as well as UGG by glyT(SuUGG-8) tRNA is unaffected whereas suppression of UGG by glyT(SuUGA/G) or glyV(SuUGA/G) tRNA is decreased in the presence of an rpsD mutation. The effects on suppressor tRNA are thus not correlated with the ribosomal ambiguity (Ram) phenotype of the rpsD mutants used in this study. It is suggested that the mutationally altered ribosomes are changed in functional interactions with the suppressor tRNA itself rather than with the competing translational release factor(s) or cognate aminoacyl tRNA. The structure of suppressor tRNA, particularly the anticodon loop, and the suppressed codon as well as the codon context determine the allele specific functional interactions with these ribosomal mutations.

This is a preview of subscription content, log in to check access.


  1. Andersson DI, Andersson SGE, Kurland CG (1986) Functional interactions between mutated forms of ribosomal protein S4, S5 and S12. Biochimie, in press

  2. Andersson DI, Bohman K, Isaksson LA, Kurland CG (1982) Translation rates and misreading characteristics of rpsD mutants in Escherichia coli. Mol Gen Genet 187:467–472

  3. Andersson DI, Kurland CG (1983) Ram ribosomes are defective proofreaders. Mol Gen Genet 191:378–381

  4. Akaboshi E, Inouye M, Tsugita A (1976) Effect of neighbouring nucleotide sequences on suppression efficiency in amber mutants of T4 phage lysozyme. Mol Gen Genet 149:1–4

  5. Biswas DK, Gorini L (1972) Restriction, de-restriction and mistranslation in missense suppression. Ribosomal discrimination of transfer RNAs. J Mol Biol 64:119–134

  6. Bohman K, Ruusala T, Jelenc PC, Kurland CG (1984) Kinetic impairment of restrictive streptomycin resistant bacteria. Mol Gen Genet 198:90–99

  7. Bossi L (1983) Context effects: Translation of UAG codons by suppressor tRNA is affected by the sequence following in the message. J Mol Biol 164:73–87

  8. Bossi L, Roth JR (1980) The influence of codon context on genetic code translation. Nature 286:123–127

  9. Bouadloun F, Donner D, Kurland CG (1983) Codon-specific missense errors in vivo. EMBO J 2:1351–1356

  10. Bouadloun F, Srichaiyo T, Isaksson LA, Björk GR (1986) Codon context sensitivity of translational suppression and accuracy is influenced by modification next to the anticodon in tRNA. J Bacteriol, in press

  11. Buck M, Griffiths E (1981) Regulation of aromatic amino acid transport by tRNA: role of 2-methylthio-N6(σ 2-isopentenyl) adenosine. Nucleic Acids Res 9:401–414

  12. Buckingham RH, Kurland CG (1977) Codon specificity of UGA suppressor tRNATrp from Escherichia coli. Proc Natl Acad Sci USA 74:5496–5498

  13. Cabezon T, Herzog A, De Wilde M, Villarroel R, Bollen A (1976) Cooperative control of translational fidelity by ribosomal proteins in Escherichia coli III. A ram mutation in the structural gene for protein S5 (rpxE). Mol Gen Genet 144:59–62

  14. Carbon J, Squires C, Hill CW (1970) Glycine transfer RNA of Escherichia coli. II. Impaired GGA-recognition in strains containing a genetically altered transfer RNA; Reversal by a secondary suppressor mutation. J Mol Biol 52:571–584

  15. Caskey CT, Forrester WL, Tate W, Ward C (1984) Cloning of the Escherichia coli release factor 2 gene. J Bacteriol 158:365–368

  16. Creighton TE, Yanofsky C (1966) Association of the α and β2 subunits of the tryptophan synthetase of Escherichia coli. J Biol Chem 241:980–990

  17. Feinstein SI, Altman S (1978) Context effects on nonsense codon suppressors in Escherichia coli. Genetics 88:201–219

  18. Fluck MM, Salser W, Epstein RH (1977) The influence of the reading context upon the suppression of nonsense codons. Mol Gen Genet 151:137–149

  19. Fluck MM, Epstein RH (1980) Isolation and characterization of context mutations affecting the suppressibility of nonsense mutations. Mol Gen Genet 177:615–627

  20. Galas JD, Branscomb WB (1976) Ribosome slowed by mutation to streptomycin resistance. Nature 262:617–619

  21. Goldberg ME, Creighton TE, Baldwin RL, Yanofsky C (1966) Subunit structure of the tryptophan synthetase of Escherichia coli. J Mol Biol 21:71–82

  22. Gorini L (1971) Ribosomal discrimination of tRNAs. Nature New Biol 234:261–264

  23. Gorini L (1974) In: Nomura M, Tissieres A, Lengyel P (eds) Ribosomes. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 791–803

  24. Hill CW, Squires C, Carbon J (1970) Glycine transfer RNA of Escherichia coli I. Structural genes for two glycine tRNA species. J Mol Biol 52:557–569

  25. Hirsch D (1971) Tryptophan transfer RNA as the UGA suppressor. J Mol Biol 58:439–458

  26. Hopfield JJ (1974) Kinetic proofreading: A new mechanism for reducing errors in biosynthetic processes requiring high specificity. Proc Natl Acad Sci USA 71:4135–4139

  27. Jackson DA, Yanofsky C (1969) The formation and properties of dimers of the α tryptophan synthetase subunit of Escherichia coli. J Biol Chem 244:4526–4538

  28. Kirsebom LA, Isaksson LA (1985) Involvement of ribosomal protein L7/L12 in control of translational accuracy. Proc Natl Acad Sci USA 82:717–721

  29. Kirsebom LA, Amons R, Isaksson LA (1986) Primary structures of mutationally altered ribosomal protein L7/L12 and their effects on cellular growth and translational accuracy. Eur J Biochem 156:669–675

  30. Kühberger R, Piepersberg W, Petzet A, Buckel P, Böck A (1979) Alteration of ribosomal protein L6 in gentamycin-resistant strains of Escherichia coli. Effects of fidelity of protein synthesis. Biochemistry 18:187–193

  31. Kurland CG, Rigler R, Ehrenberg M, Blomberg C (1975) Allosteric mechanism for codon dependent tRNA selection on ribosomes. Proc Natl Acad Sci USA 72:4248–4251

  32. Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York

  33. Miller JH, Albertini AM (1983) Effects of surrounding sequence on the suppression of nonsense codons. J Mol Biol 164:59–71

  34. Miller JH, Coulondre C, Farabough PJ (1978) Correlation of nonsense sites in the lacI gene with specific codons in the nucleotide sequence. Nature 274:770–775

  35. Murgola EJ (1981) Restricted wobble in UGA codon recognition by glycine tRNA suppressors of UGA. J Mol Biol 149:1–13

  36. Murgola EJ (1985) tRNA, suppression and the code. Annu Rev Genet 19:57–80

  37. Murgola EJ, Childress JR (1980) Suppressors of a UGG missense mutation in Escherichia coli. J Bacteriol 143:285–292

  38. Murgola EJ, Jones CI (1978) A novel method for detection and characterization of ochre suppressors in Escherichia coli. Mol Gen Genet 159:179–184

  39. Murgola EJ, Pagel FT (1983) Suppressors of lysine codons may be misacylated lysine tRNAs. J Bacteriol 156:917–919

  40. Murgola EJ, Pagel FT, Hijazi KA (1984) Codon context effects in missense suppression. J Mol Biol 175:19–27

  41. Murgola EJ, Prather NE, Mims BH, Pagel FT, Hijazi K (1983) Anticodon shift in tRNA: A novel mechanism in missense and nonsense suppression. Proc Natl Acad Sci USA 80:4936–4939

  42. Neidhardt FC, Bloch PL, Smith DF (1974) Culture medium for Enterobacteria. J Bacteriol 119:736–747

  43. Ninio J (1974) A semi-quantitative treatment of missense and nonsense suppression in the strA and ram ribosomal mutants of Escherichia coli. J Mol Biol 84:297–313

  44. Ninio J (1975) Kinetic amplification of enzyme discrimination. Biochimie 57:587–595

  45. O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021

  46. Olsson MO, Isaksson LA (1979) Analysis of rpsD mutations in Escherichia coli. I. Comparison of mutants with various alterations in ribosomal protein S4. Mol Gen Genet 169:251–257

  47. Olsson MO, Isaksson LA (1980) Analysis of rpsD mutations in Escherichia coli. IV. Accumulation of minor forms of protein S7(K) in ribosomes of rpsD mutant strains due to translational read-through. Mol Gen Genet 177:485–491

  48. Ozeki H, Inokuchi H, Yamao F, Kodaira M, Sakano H, Ikemura T, Shimura Y (1980) In: Söll D, Abelson JN, Schimmel PR (eds) Transfer RNA: Biological aspects. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 341–362

  49. Piepersberg W, Böck A, Wittman HG (1975) Effect of different mutations in ribosomal protein S5 of Escherichia coli on translational fidelity. Mol Gen Genet 140:271–278

  50. Prather NE, Murgola EJ, Mims BH (1981a) Primary structure of an unusual glycine tRNA UGA suppressor. Nucleic Acids Res 9:6421–6428

  51. Prather NE, Murgola EJ, Mims B (1981b) Nucleotide insertion in the anticodon loop of a glycine transfer RNA causes missense suppression. Proc Natl Acad Sci USA 78:7408–7411

  52. Roberts JW, Carbon J (1975) Nucleotide sequence studies of normal and genetically altered glycine transfer ribonucleic acids from Escherichia coli. J Biol Chem 250:5530–5541

  53. Rosset R, Gorini L (1969) A ribosomal ambiguity mutation. J Mol Biol 39:95–112

  54. Ruusala T, Ehrenberg M, Kurland CG (1982) Is there proofreading during polypeptide synthesis? EMBO J 1:741–745

  55. Ruusala T, Andersson D, Ehrenberg M, Kurland CG (1984) Hyperaccurate ribosomes inhibit growth. EMBO J 3:2575–2580

  56. Salser W (1969) The influence of the reading codon context upon the suppression of nonsense codons. Mol Gen Genet 105:125–130

  57. Strigini P, Gorini L (1970) Ribosomal mutations affecting efficiency of amber supression. J Mol Biol 47:517–530

  58. Thompson RC, Stone PJ (1977) Proofreading of the codon-anticodon interaction on ribosomes. Proc Natl Acad Sci USA 74:198–202

  59. Thompson RC, Dix BD, Gerson BR, Karim MA (1981) Effect of Mg2+ concentration, polyamines, streptomycin, and mutations in ribosomal proteins on the accuracy of the two-step selection of aminoacyl-tRNAs in protein biosynthesis. J Biol Chem 256:6676–6681

  60. Vogel HJ, Bonner DM (1956) Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem 218:97–106

  61. Weiss RB, Murphy JP, Gallant JA (1984) Genetic screen for cloned release factor genes. J Bacteriol 158:362–364

  62. Yanofsky C, Platt T, Crawford IP, Nichols BP, Christie GE, Horowitz H, Van Cleemput M (1981) The complete nucleotide sequence of the tryptophan operon of Escherichia coli. Nucleic Acids Res 9:6647–6668

  63. Yahata H, Ocada Y, Tsugita A (1970) Adjacent effect on suppression efficiency. II. Study on ochre and amber mutants of T4 phage lysozyme. Mol Gen Genet 106:208–212

  64. Yarus M (1982) Translation efficiency of transfer RNA's: Uses of an extended anticodon. Science 218:646–652

  65. Yarus M, McMillan C, Cline S, Bradley D, Snyder M (1980) Construction of a composite tRNA gene by anticodon loop transplant. Proc Natl Acad Sci USA 77:5092–5096

  66. Yates JL (1979) Role of ribosomal protein S12 in discrimination of aminoacyl-tRNA. J Biol Chem 254:11550–11554

  67. Zimmerman RA, Garvin T, Gorin L (1971) Alteration of a 30S ribosomal protein accompanying the ram mutation in Escherichia coli. Proc Natl Acad Sci USA 68:2263–2267

Download references

Author information

Correspondence to Leif A. Isaksson.

Additional information

Communicated by A. Böck

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kirsebom, L.A., Isaksson, L.A. Functional interactions in vivo between suppressor tRNA and mutationally altered ribosomal protein S4. Molec. Gen. Genet. 205, 240–247 (1986).

Download citation

Key words

  • Ribosomal-ambiguity
  • rpsD
  • Suppressor tRNA
  • Codon context
  • in vivo