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. 1973 Oct;70(10):2870–2873. doi: 10.1073/pnas.70.10.2870

ATPase and GTPase Activities Associated with a Specific 5S RNA-Protein Complex*

Janet R Horne 1, Volker A Erdmann 1,†
PMCID: PMC427128  PMID: 4355369

Abstract

An RNA-protein complex consisting of 5S RNA and two ribosomal proteins, B-L5 and B-L22, was isolated from Bacillus stearothermophilus ribosomes and found to be active in GTP hydrolysis. This activity was not influenced by elongation factor G. Further analysis of this complex showed that it was also able to hydrolyze ATP. Inhibition studies revealed that ATP was a noncompetitive inhibitor of GTP and that GTP was also a noncompetitive inhibitor of ATP, indicating that two different enzymatic sites were involved. Differences in pH optimum and optimal temperature also point to a twosite enzyme complex. Both enzymatic hydrolyses were inhibited by thiostrepton and fusidic acid, which are known inhibitors of protein synthesis.

Keywords: B. stearothermophilus, 50S ribosomal proteins, fusidic acid, thiostrepton, reconstitution

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Selected References

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

  1. Bodley J. W., Lin L., Highland J. H. Studies on translocation. VI. Thiostrepton prevents the formation of a ribosome-G factor-guanine nucleotide complex. Biochem Biophys Res Commun. 1970 Dec 24;41(6):1406–1411. doi: 10.1016/0006-291x(70)90543-7. [DOI] [PubMed] [Google Scholar]
  2. Bodley J. W., Zieve F. J., Lin L. Studies on translocation. IV. The hydrolysis of a single round of guanosine triphosphate in the presence of fusidic acid. J Biol Chem. 1970 Nov 10;245(21):5662–5667. [PubMed] [Google Scholar]
  3. CONWAY T. W., LIPMANN F. CHARACTERIZATION OF A RIBOSOME-LINKED GUANOSINE TRIPHOSPHATASE IN ESCHERICHIA COLI EXTRACTS. Proc Natl Acad Sci U S A. 1964 Dec;52:1462–1469. doi: 10.1073/pnas.52.6.1462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cabrer B., Vázquez D., Modolell J. Inhibition by elongation factor EF G of aminoacyl-tRNA binding to ribosomes. Proc Natl Acad Sci U S A. 1972 Mar;69(3):733–736. doi: 10.1073/pnas.69.3.733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cashel M., Gallant J. Two compounds implicated in the function of the RC gene of Escherichia coli. Nature. 1969 Mar 1;221(5183):838–841. doi: 10.1038/221838a0. [DOI] [PubMed] [Google Scholar]
  6. Cashel M. The control of ribonucleic acid synthesis in Escherichia coli. IV. Relevance of unusual phosphorylated compounds from amino acid-starved stringent strains. J Biol Chem. 1969 Jun 25;244(12):3133–3141. [PubMed] [Google Scholar]
  7. Cundliffe E. The mode of action of fusidic acid. Biochem Biophys Res Commun. 1972 Mar 10;46(5):1794–1801. doi: 10.1016/0006-291x(72)90053-8. [DOI] [PubMed] [Google Scholar]
  8. Cundliffe E. The mode of action of thiostreption in vivo. Biochem Biophys Res Commun. 1971 Aug 20;44(4):912–917. doi: 10.1016/0006-291x(71)90798-4. [DOI] [PubMed] [Google Scholar]
  9. Erdmann V. A., Doberer H. G. Structure and function of 5S RNA: the role of the 3' terminus in 5S RNA function. Mol Gen Genet. 1972;114(2):89–94. doi: 10.1007/BF00332779. [DOI] [PubMed] [Google Scholar]
  10. Gordon J. Hydrolysis of guanosine 5'-triphosphate associated wh binding of aminoacyl transfer ribonucleic acid to ribosomes. J Biol Chem. 1969 Oct 25;244(20):5680–5686. [PubMed] [Google Scholar]
  11. Hafkenscheid J. C., Bonting S. L. Studies on (Na+-K+)-activated ATPase. XIX. Occurrence and properties of a (Na+-K+)-activated ATPase in Escherichia coli. Biochim Biophys Acta. 1968 Jan 8;151(1):204–211. doi: 10.1016/0005-2744(68)90175-7. [DOI] [PubMed] [Google Scholar]
  12. Haseltine W. A., Block R., Gilbert W., Weber K. MSI and MSII made on ribosome in idling step of protein synthesis. Nature. 1972 Aug 18;238(5364):381–384. doi: 10.1038/238381a0. [DOI] [PubMed] [Google Scholar]
  13. Horne J. R., Erdmann V. A. Isolation and characterization of 5S RNA-protein complexes from Bacillus stearothermophilus and Escherichia coli ribosomes. Mol Gen Genet. 1972;119(4):337–344. doi: 10.1007/BF00272091. [DOI] [PubMed] [Google Scholar]
  14. Kaziro Y., Inoue N., Kuriki Y., Mizumoto K., Tanaka M., Kawakita M. Purification and properties of factor G. Cold Spring Harb Symp Quant Biol. 1969;34:385–393. doi: 10.1101/sqb.1969.034.01.045. [DOI] [PubMed] [Google Scholar]
  15. Kurland C. G. Structure and function of the bacterial ribosome. Annu Rev Biochem. 1972;41(10):377–408. doi: 10.1146/annurev.bi.41.070172.002113. [DOI] [PubMed] [Google Scholar]
  16. Lucas-Lenard J. Protein biosynthesis. Annu Rev Biochem. 1971;40:409–448. doi: 10.1146/annurev.bi.40.070171.002205. [DOI] [PubMed] [Google Scholar]
  17. Lucas-Lenard J., Tao P., Haenni A. L. Further studies on bacterial polypeptide elongation. Cold Spring Harb Symp Quant Biol. 1969;34:455–462. doi: 10.1101/sqb.1969.034.01.051. [DOI] [PubMed] [Google Scholar]
  18. Lund E., Kjeldgaard N. O. Metabolism of guanosine tetraphosphate in Escherichia coli. Eur J Biochem. 1972 Jul 24;28(3):316–326. doi: 10.1111/j.1432-1033.1972.tb01916.x. [DOI] [PubMed] [Google Scholar]
  19. Miller D. L. Elongation factors EF Tu and EF G interact at related sites on ribosomes. Proc Natl Acad Sci U S A. 1972 Mar;69(3):752–755. doi: 10.1073/pnas.69.3.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Modolell J., Cabrer B., Parmeggiani A., Vazquez D. Inhibition by siomycin and thiostrepton of both aminoacyl-tRNA and factor G binding to ribosomes. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1796–1800. doi: 10.1073/pnas.68.8.1796. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Modolell J., Vazquez D., Monro R. E. Ribosomes, G-factor and siomycin. Nat New Biol. 1971 Mar 24;230(12):109–112. doi: 10.1038/newbio230109a0. [DOI] [PubMed] [Google Scholar]
  22. Nishizuka Y., Lipmann F. The interrelationship between guanosine triphosphatase and amino acid polymerization. Arch Biochem Biophys. 1966 Sep 26;116(1):344–351. doi: 10.1016/0003-9861(66)90040-3. [DOI] [PubMed] [Google Scholar]
  23. Nomura M. Assembly of bacterial ribosomes. Science. 1973 Mar 2;179(4076):864–873. doi: 10.1126/science.179.4076.864. [DOI] [PubMed] [Google Scholar]
  24. Nomura M., Erdmann V. A. Reconstitution of 50S ribosomal subunits from dissociated molecular components. Nature. 1970 Nov 21;228(5273):744–748. doi: 10.1038/228744a0. [DOI] [PubMed] [Google Scholar]
  25. Ono Y., Skoultchi A., Waterson J., Lengyel P. Peptide chain elongation: GTP cleavage catalysed by factors binding aminoacyl-transfer RNA to the ribosome. Nature. 1969 May 17;222(5194):645–648. doi: 10.1038/222645a0. [DOI] [PubMed] [Google Scholar]
  26. Pedersen F. S., Lund E., Kjeldgaard N. O. Codon specific, tRNA dependent in vitro synthesis of ppGpp and pppGpp. Nat New Biol. 1973 May 2;243(122):13–15. [PubMed] [Google Scholar]
  27. Pestka S. Thiostrepton: a ribosomal inhibitor of translocation. Biochem Biophys Res Commun. 1970 Aug 11;40(3):667–674. doi: 10.1016/0006-291x(70)90956-3. [DOI] [PubMed] [Google Scholar]
  28. Richman N., Bodley J. W. Ribosomes cannot interact simultaneously with elongation factors EF Tu and EF G. Proc Natl Acad Sci U S A. 1972 Mar;69(3):686–689. doi: 10.1073/pnas.69.3.686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Richter D. Inability of E. coli ribosomes to interact simultaneously with the bacterial elongation factors EF Tu and EF G. Biochem Biophys Res Commun. 1972 Mar 10;46(5):1850–1856. doi: 10.1016/0006-291x(72)90061-7. [DOI] [PubMed] [Google Scholar]
  30. Richter D., Lin L., Bodley J. W. Studies on translocation IX. The pattern of action of antibiotic translocation inhibitors in eukaryotic and prokaryotic systems. Arch Biochem Biophys. 1971 Nov;147(1):186–191. doi: 10.1016/0003-9861(71)90326-2. [DOI] [PubMed] [Google Scholar]
  31. Roisin M. P., Kepes A. The membrane ATPase of Escherichia coli. I. Ion dependence and ATP-ADP exchange reaction. Biochim Biophys Acta. 1972 Sep 20;275(3):333–346. doi: 10.1016/0005-2728(72)90214-9. [DOI] [PubMed] [Google Scholar]
  32. Shorey R. L., Ravel J. M., Garner C. W., Shive W. Formation and properties of the aminoacyl transfer ribonucleic acid-guanosine triphosphate-protein complex. J Biol Chem. 1969 Sep 10;244(17):4555–4564. [PubMed] [Google Scholar]
  33. Sy J., Lipmann F. Identification of the synthesis of guanosine tetraphosphate (MS I) as insertion of a pyrophosphoryl group into the 3'-position in guanosine 5'-diphosphate. Proc Natl Acad Sci U S A. 1973 Feb;70(2):306–309. doi: 10.1073/pnas.70.2.306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tanaka N., Kinoshita T., Masukawa H. Mechanism of protein synthesis inhibition by fusidic acid and related antibiotics. Biochem Biophys Res Commun. 1968 Feb 15;30(3):278–283. doi: 10.1016/0006-291x(68)90447-6. [DOI] [PubMed] [Google Scholar]
  35. Weisblum B., Demohn V. Thiostrepton, an inhibitor of 5OS ribosome subunit function. J Bacteriol. 1970 Mar;101(3):1073–1075. doi: 10.1128/jb.101.3.1073-1075.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]

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