Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1977 Jul;4(7):2205–2212. doi: 10.1093/nar/4.7.2205

Properties of phenylalanine transfer ribonucleic acid with modified 3'-terminal end in protein biosynthesis using a rabbit reticulocyte cell-free system: effect of the replacement of cytidine residues from the CpCpA end of tRNA by 5-iodocytidine or 2-thiocytidine.

E K Baksht, A Gal, N de Groot, A A Hochberg, M Sprinzl, F Cramer
PMCID: PMC342559  PMID: 242797

Abstract

Phe-tRNA Phe from yeast containing 2-thiocytidine or 5-iodocytidine in position 75 of the polynucleotide chain or Phe-tRNA Phe in which both positions 74 and 75 were substituted by 5-iodocytidine were investigated in the poly U-dependent polyphenylalanine synthesis on ribosomes from rabbit reticulocytes. Phe-tRNA Phe-Cps2CpA was nearly as active as the native Phe-tRNA Phe-CpCpA in the overall process. Phe-tRNA Phe-Cpi 5CpA as well as Phe-tRNA Phe-i5Cpi 5CpA were considerably less active than the native species. Investigation of individual steps of protein biosynthesis with these modified substrates revealed that the donor activity of peptidyl-tRNAs which contain 5-iodocytidine in their 3'-terminus is strongly imparied suggesting exacting structural requirements for the interaction of the CpCpA end of tRNA with the ribosomal P-site.

Full text

PDF
2205

Selected References

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

  1. Baksht E., de Groot N. D. The enzymatic binding of aminoacyl-tRNA to reticulocyte ribosomes: the stimulatory effect of donor site bound peptidyl-tRNA. Mol Biol Rep. 1974 Dec;1(8):493–497. doi: 10.1007/BF00360677. [DOI] [PubMed] [Google Scholar]
  2. Baksht E., de Groot N., Sprinzl M., Cramer F. Properties of tRNA species modified in the 3'-terminal ribose moiety in an eukaryotic ribosomal system. Biochemistry. 1976 Aug 10;15(16):3639–3646. doi: 10.1021/bi00661a035. [DOI] [PubMed] [Google Scholar]
  3. Baksht E., de Groot N., Sprinzl M., Cramer F. The behaviour of phenylalanine transfer ribonucleic acid with 3'-terminal formycin in protein biosynthesis using a rabbit reticulocyte cell-free system. FEBS Lett. 1975 Jul 15;55(1):105–108. doi: 10.1016/0014-5793(75)80970-7. [DOI] [PubMed] [Google Scholar]
  4. Chinali G., Sprinzl M., Parmeggiani A., Cramer F. Participation in protein biosynthesis of transfer ribonucleic acids bearing altered 3'-terminal ribosyl residues. Biochemistry. 1974 Jul 16;13(15):3001–3010. doi: 10.1021/bi00712a001. [DOI] [PubMed] [Google Scholar]
  5. Deutscher M. P. Synthesis and functions of the -C-C-A terminus of transfer RNA. Prog Nucleic Acid Res Mol Biol. 1973;13:51–92. doi: 10.1016/s0079-6603(08)60100-2. [DOI] [PubMed] [Google Scholar]
  6. Eckermann D. J., Greenwell P., Symons R. H. Peptide-bond formation on the ribosome. A comparison of the acceptor-substrate specificity of peptidyl transferase in bacterial and mammalian ribosomes using puromycin analogues. Eur J Biochem. 1974 Feb 1;41(3):547–554. doi: 10.1111/j.1432-1033.1974.tb03296.x. [DOI] [PubMed] [Google Scholar]
  7. Faerber P., Scheit K. H., Sommer H. A new polynucleotide complex poly(s 2 C)-poly(I). Eur J Biochem. 1972 May;27(1):109–115. doi: 10.1111/j.1432-1033.1972.tb01816.x. [DOI] [PubMed] [Google Scholar]
  8. Fraser T. H., Rich A. Synthesis and aminoacylation of 3'-amino-3'-deoxy transfer RNA and its activity in ribosomal protein synthesis. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2671–2675. doi: 10.1073/pnas.70.9.2671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hecht S. M., Kozarich J. W., Schmidt F. J. Isomeric phenylalanyl-tRNAs. Position of the aminoacyl moiety during protein biosynthesis. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4317–4321. doi: 10.1073/pnas.71.11.4317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Leder P., Bursztyn H. Initiation of protein synthesis II. A convenient assay for the ribosome-dependent synthesis of N-formyl-C14-methionylpuromycin. Biochem Biophys Res Commun. 1966 Oct 20;25(2):233–238. doi: 10.1016/0006-291x(66)90586-9. [DOI] [PubMed] [Google Scholar]
  11. Monro R. E., Cerná J., Marcker K. A. Ribosome-catalyzed peptidyl transfer: substrate specificity at the P-site. Proc Natl Acad Sci U S A. 1968 Nov;61(3):1042–1049. doi: 10.1073/pnas.61.3.1042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. NIRENBERG M., LEDER P. RNA CODEWORDS AND PROTEIN SYNTHESIS. THE EFFECT OF TRINUCLEOTIDES UPON THE BINDING OF SRNA TO RIBOSOMES. Science. 1964 Sep 25;145(3639):1399–1407. doi: 10.1126/science.145.3639.1399. [DOI] [PubMed] [Google Scholar]
  13. Ofengand J., Chen C. M. Inactivation of T u factor-guanosine triphosphate recognition and ribosome-binding ability by terminal oxidation-reduction of yeast phenylalanine transfer ribonucleic acid. J Biol Chem. 1972 Apr 10;247(7):2049–2058. [PubMed] [Google Scholar]
  14. Rappoport S., Lapidot Y. The chemical preparation of acetylaminoacyl-tRNA. Methods Enzymol. 1974;29:685–688. doi: 10.1016/0076-6879(74)29060-8. [DOI] [PubMed] [Google Scholar]
  15. Sneden D., Miller D. L., Kim S. H., Rich A. Preliminary x-ray analysis of the crystalline complex between poypeptide chain elongation factor, Tu, and GDP. Nature. 1973 Feb 23;241(5391):530–531. doi: 10.1038/241530a0. [DOI] [PubMed] [Google Scholar]
  16. Sprinzl M., Cramer F. Accepting site for aminoacylation of tRNAphe from yeast. Nat New Biol. 1973 Sep 5;245(140):3–5. doi: 10.1038/newbio245003a0. [DOI] [PubMed] [Google Scholar]
  17. Sprinzl M., Scheit K. H., Cramer F. Preparation in vitro of a 2-thiocytidine-containing yeast tRNA Phe -A 73 -C 74 -S 2 C 75 -A 76 and its interaction wiith p-hydroxymercuribenzoate. Eur J Biochem. 1973 Apr;34(2):306–310. doi: 10.1111/j.1432-1033.1973.tb02759.x. [DOI] [PubMed] [Google Scholar]
  18. Sprinzl M., von der Haar F., Schlimme E., Sternbach H., Cramer F. Incorporation of 5-iodocytidine into yeast tRNAphe with tRNA nucleotidyl transferase in vitro. Eur J Biochem. 1972 Feb 15;25(2):262–266. doi: 10.1111/j.1432-1033.1972.tb01692.x. [DOI] [PubMed] [Google Scholar]
  19. Watanabe K., Oshima T., Nishimura S. CD spectra of 5-methyl-2-thiouridine in tRNA-Met-f from an extreme thermophile. Nucleic Acids Res. 1976 Jul;3(7):1703–1713. doi: 10.1093/nar/3.7.1703. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES