Skip to main content
NCBI home page
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
. 1976 Jun;73(6):1839–1843. doi: 10.1073/pnas.73.6.1839

Ribosomal proteins of Escherichia coli that stimulate stringent-factor-mediated pyrophosphoryl transfer in vitro.

L Christiansen, K H Neirhaus
PMCID: PMC430402  PMID: 778846

Abstract

Guanosine tetra- and pentaphosphate, (p)ppGpp, can be synthesized in vitro in a reaction containing only the enzyme (stringent factor), salts, and substrates (nonribosomal system). This reaction is greatly stimulated upon addition of methanol (methanol system) or by ribosomes, mRNA, and tRNA (ribosome system). Here we show that several ribosomal proteins alone stimulate the synthesis of (p)ppGpp in the presence of stringent factor (protein system). The optimal ionic conditions for the ribosome and protein systems are identical. The concentration of ribosomes or any stimulating ribosomal protein required for saturation of a given concentration of stringent factor is similar. Fifty of 54 ribosomal proteins were tested for stimulation in the protein system; 15 proteins showed high activity, seven of these from the 30S ribosomal subunit and eight from the 50S subunit. The physiological relevance of this finding is discussed.

Full text

PDF
1839

Images in this article

1842Image
on p.1842

Selected References

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

  1. Barritault D., Expert-Bezançon A., Milet M. Etude des relations de voisinage entre des protéines de la sous-unité ribosomique 50 S d'E. coli. C R Acad Sci Hebd Seances Acad Sci D. 1975 Oct 6;281(14):1043–1046. [PubMed] [Google Scholar]
  2. Block R., Haseltine A. W. Purification and properties of stringent factor. J Biol Chem. 1975 Feb 25;250(4):1212–1217. [PubMed] [Google Scholar]
  3. Cashel M., Kalbacher B. The control of ribonucleic acid synthesis in Escherichia coli. V. Characterization of a nucleotide associated with the stringent response. J Biol Chem. 1970 May 10;245(9):2309–2318. [PubMed] [Google Scholar]
  4. Cochran J. W., Byrne R. W. Isolation and properties of a ribosome-bound factor required for ppGpp and ppGpp synthesis in Escherichia coli. J Biol Chem. 1974 Jan 25;249(2):353–360. [PubMed] [Google Scholar]
  5. Expert-Bezançon A., Barritault D., Clegg J. C., Milet M., Khouvine Y., Hayes D. H. Identification of neighbouring proteins in E. coli 50S ribosomes. FEBS Lett. 1975 Nov 1;59(1):64–69. doi: 10.1016/0014-5793(75)80342-5. [DOI] [PubMed] [Google Scholar]
  6. Gesteland R. F. Isolation and characterization of ribonuclease I mutants of Escherichia coli. J Mol Biol. 1966 Mar;16(1):67–84. doi: 10.1016/s0022-2836(66)80263-2. [DOI] [PubMed] [Google Scholar]
  7. Hamel E., Koka M., Nakamoto T. Requirement of an Escherichia coli 50 S ribosomal protein component for effective interaction of the ribosome with T and G factors and with guanosine triphosphate. J Biol Chem. 1972 Feb 10;247(3):805–814. [PubMed] [Google Scholar]
  8. Hardy S. J. The stoichiometry of the ribosomal proteins of Escherichia coli. Mol Gen Genet. 1975 Oct 3;140(3):253–274. doi: 10.1007/BF00334270. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Haseltine W. A., Block R. Synthesis of guanosine tetra- and pentaphosphate requires the presence of a codon-specific, uncharged transfer ribonucleic acid in the acceptor site of ribosomes. Proc Natl Acad Sci U S A. 1973 May;70(5):1564–1568. doi: 10.1073/pnas.70.5.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hindennach I., Kaltschmidt E., Wittmann H. G. Ribosomal proteins. Isolation of proteins from 50S ribosomal subunits of Escherichia coli. Eur J Biochem. 1971 Nov 11;23(1):12–16. doi: 10.1111/j.1432-1033.1971.tb01585.x. [DOI] [PubMed] [Google Scholar]
  12. Hindennach I., Stöffler G., Wittmann H. G. Ribosomal proteins. Isolation of the proteins from 30S ribosomal subunits of Escherichia coli. Eur J Biochem. 1971 Nov 11;23(1):7–11. doi: 10.1111/j.1432-1033.1971.tb01584.x. [DOI] [PubMed] [Google Scholar]
  13. Homann H. E., Nierhaus K. H. Ribosomal proteins. Protein compositions of biosynthetic precursors and artifical subparticles from ribosomal subunits in Escherichia coli K 12. Eur J Biochem. 1971 May 28;20(2):249–257. doi: 10.1111/j.1432-1033.1971.tb01388.x. [DOI] [PubMed] [Google Scholar]
  14. Kaltschmidt E. Ribosomal proteins. XIV. Isoelectric points of ribosomal proteins of E. coli as determined by two-dimensional polyacrylamide gel electrophoresis. Anal Biochem. 1971 Sep;43(1):25–31. doi: 10.1016/0003-2697(71)90103-5. [DOI] [PubMed] [Google Scholar]
  15. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  16. Nierhaus K. H., Dohme F. Total reconstitution of functionally active 50S ribosomal subunits from Escherichia coli. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4713–4717. doi: 10.1073/pnas.71.12.4713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nierhaus K. H., Montejo V. A protein involved in the peptidyltransferase activity of Escherichia coli ribosomes. Proc Natl Acad Sci U S A. 1973 Jul;70(7):1931–1935. doi: 10.1073/pnas.70.7.1931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Parker J., Watson R. J., Friesen J. D. A relaxed mutant with an altered ribosomal protein L11. Mol Gen Genet. 1976 Feb 27;144(1):111–114. doi: 10.1007/BF00277313. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Reiness G., Yang H. L., Zubay G., Cashel M. Effects of guanosine tetraphosphate on cell-free synthesis of Escherichia coli ribosomal RNA and other gene products. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2881–2885. doi: 10.1073/pnas.72.8.2881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Richter D., Nowak P., Kleinert U. Escherichia coli stringent factor binds to ribosomes at a site different from that of elongation factor Tu or G. Biochemistry. 1975 Oct 7;14(20):4414–4420. doi: 10.1021/bi00691a012. [DOI] [PubMed] [Google Scholar]
  22. Rohde M. F., Aune K. C. Molecular interactions between ribosomal proteins. Evidence for specificity of interaction between isolated proteins. Biochemistry. 1975 Sep 23;14(19):4344–4348. doi: 10.1021/bi00690a031. [DOI] [PubMed] [Google Scholar]
  23. Rohde M. F., O'Brien S., Cooper S., Aune K. C. Physical properties of some ribosomal proteins in solution and evidence for molecular interactions between isolated ribosomal proteins. Biochemistry. 1975 Mar 11;14(5):1079–1087. doi: 10.1021/bi00676a031. [DOI] [PubMed] [Google Scholar]
  24. Schreiner G., Nierhaus K. H. Protein involved in the binding of dihydrostreptomycin to ribosomes of Escherichia coli. J Mol Biol. 1973 Nov 25;81(1):71–82. doi: 10.1016/0022-2836(73)90248-9. [DOI] [PubMed] [Google Scholar]
  25. Sy J., Ogawa Y., Lipmann F. Nonribosomal synthesis of guanosine 5',3'-polyphosphates by the ribosomal wash of stringent Escherichia coli. Proc Natl Acad Sci U S A. 1973 Jul;70(7):2145–2148. doi: 10.1073/pnas.70.7.2145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tischendorf G. W., Zeichhardt H., Stöffler G. Architecture of the Escherichia coli ribosome as determined by immune electron microscopy. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4820–4824. doi: 10.1073/pnas.72.12.4820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ulbrich B., Nierhaus K. H. Pools of ribosomal proteins in Escherichia coli. Studies on the exchange of proteins between pools and ribosomes. Eur J Biochem. 1975 Sep 1;57(1):49–54. doi: 10.1111/j.1432-1033.1975.tb02275.x. [DOI] [PubMed] [Google Scholar]
  28. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  29. van Ooyen A. J., de Boer H. A., Ab G., Gruber M. Specific inhibition of ribosomal RNA synthesis in vitro by guanosine 3' diphosphate, 5' diphosphate. Nature. 1975 Apr 10;254(5500):530–531. doi: 10.1038/254530a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES