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. 1973 Jan;70(1):147–150. doi: 10.1073/pnas.70.1.147

Identity of the Ribosomal Proteins Involved in the Interaction with Elongation Factor G*

Joseph H Highland †,‡, James W Bodley §, Julian Gordon , Renate Hasenbank †, Georg Stöffler †
PMCID: PMC433203  PMID: 4567331

Abstract

Rabbit antibodies produced against 50 of the 55 individually purified ribosomal proteins of Escherichia coli were tested for their ability to interfere with the formation of the ribosome·EF-G·GDP complex. Only antibodies produced against proteins L7 and L12 inhibited complex formation, and they did so completely. These two proteins were previously shown to be immunologically indistinguishable and necessary for the interaction between ribosomes and EF-G. The present data are consistent with the view that the interaction between ribosomes and EF-G that results in GTP hydrolysis occurs on, and is limited to, proteins L7 and L12 on the surface of the 50S ribosomal subunit.

Keywords: E. coli, ribosomal protein antibodies, translocation, GTP, fusidic acid

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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. Interaction of E. coli G factor with the 50S ribosomal subunit. Nature. 1970 Jul 4;227(5253):60–61. doi: 10.1038/227060a0. [DOI] [PubMed] [Google Scholar]
  2. Bodley J. W., Lin L. Studies on the nature of the G-factor binding site on the 50S ribosomal subunit. Biochemistry. 1972 Feb 29;11(5):782–786. doi: 10.1021/bi00755a016. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Bodley J. W., Zieve F. J., Lin L., Zieve S. T. Studies on translocation. 3. Conditions necessary for the formation and detection of a stable ribosome-G factor-guanosine diphosphate complex in the presence of fusidic acid. J Biol Chem. 1970 Nov 10;245(21):5656–5661. [PubMed] [Google Scholar]
  5. Brot N., Yamasaki E., Redfield B., Weissbach H. The properties of an E. coli ribosomal protein required for the function of factor G. Arch Biochem Biophys. 1972 Jan;148(1):148–155. doi: 10.1016/0003-9861(72)90125-7. [DOI] [PubMed] [Google Scholar]
  6. 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]
  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. Deusser E., Wittmann H. G. Ribosomal proteins: variation of the protein composition in Escherichia coli ribosomes as function of growth rate. Nature. 1972 Aug 4;238(5362):269–270. doi: 10.1038/238269a0. [DOI] [PubMed] [Google Scholar]
  9. Dzionara M. Ribosomal proteins. Secondary structure of individual ribosomal proteins of E. coli studied by circular dichroism. FEBS Lett. 1970 Jun 8;8(4):197–200. doi: 10.1016/0014-5793(70)80262-9. [DOI] [PubMed] [Google Scholar]
  10. Gray P. N., Garrett R. A., Stoffler G., Monier R. An attempt at the identification of the proteins involved in the incorporation of 5-S RNA during 50-S ribosomal subunit assembly. Eur J Biochem. 1972 Jul 24;28(3):412–421. doi: 10.1111/j.1432-1033.1972.tb01927.x. [DOI] [PubMed] [Google Scholar]
  11. Grunberg-Manago M., Dondon J., Graffe M. Inhibition by thiostrepton of the IF-2-dependent ribosomal GTPase. FEBS Lett. 1972 May 1;22(2):217–221. doi: 10.1016/0014-5793(72)80049-8. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Highland J. H., Lin L., Bodley J. W. Protection of ribosomes from thiostrepton inactivation by the binding of G factor and guanosine diphosphate. Biochemistry. 1971 Nov 23;10(24):4404–4409. doi: 10.1021/bi00800a009. [DOI] [PubMed] [Google Scholar]
  14. Kaltschmidt E., Dzionara M., Wittmann H. G. Ribosomal proteins. XV. Amino acid compositions of isolated ribosomal proteins from 30S and 50S subunits of Escherichia coli. Mol Gen Genet. 1970;109(4):292–297. doi: 10.1007/BF00267698. [DOI] [PubMed] [Google Scholar]
  15. Kischa K., Möller W., Stöffler G. Reconstitution of a GTPase activity by a 50S ribosomal protein and E. coli. Nat New Biol. 1971 Sep 8;233(36):62–63. doi: 10.1038/newbio233062a0. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Lucas-Lenard J. Protein biosynthesis. Annu Rev Biochem. 1971;40:409–448. doi: 10.1146/annurev.bi.40.070171.002205. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Mora G., Donner D., Thammana P., Lutter L., Kurland C. G., Craven G. R. Purification and characterization of 50S ribosomal proteins of Escherichia coli. Mol Gen Genet. 1971;112(3):229–242. doi: 10.1007/BF00269176. [DOI] [PubMed] [Google Scholar]
  21. Möller W., Castleman H., Terhorst C. P. Characterization of an acidic protein in 50 s ribosomes of E. coli. FEBS Lett. 1970 Jun 8;8(4):192–196. doi: 10.1016/0014-5793(70)80261-7. [DOI] [PubMed] [Google Scholar]
  22. Möller W., Groene A., Terhorst C., Amons R. 50-S ribosomal proteins. Purification and partial characterization of two acidic proteins, A 1 and A 2, isolated from 50-S ribosomes of Escherichia coli. Eur J Biochem. 1972 Jan 31;25(1):5–12. doi: 10.1111/j.1432-1033.1972.tb01660.x. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Stöffler G., Daya L., Rak K. H., Garrett R. A. Ribosomal proteins. XXVI. The number of specific protein binding sites on 16 s and 23 s RNA of Escherichia coli. J Mol Biol. 1971 Dec 14;62(2):411–414. doi: 10.1016/0022-2836(71)90437-2. [DOI] [PubMed] [Google Scholar]
  26. Stöffler G., Wittmann H. G. Ribosomal proteins. XXV. Immunological studies on Escherichia coli ribosomal proteins. J Mol Biol. 1971 Dec 14;62(2):407–409. doi: 10.1016/0022-2836(71)90436-0. [DOI] [PubMed] [Google Scholar]
  27. Stöffler G., Wittmann H. G. Sequence differences of Escherichia coli 30S ribosomal proteins as determined by immunochemical methods. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2283–2287. doi: 10.1073/pnas.68.9.2283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Terhorst C., Wittmann-Liebold B., Möller W. 50-S ribosomal proteins. Peptide studies on two acidic proteins, A 1 and A 2 , isolated from 50-S ribosomes of Escherichia coli. Eur J Biochem. 1972 Jan 31;25(1):13–19. doi: 10.1111/j.1432-1033.1972.tb01661.x. [DOI] [PubMed] [Google Scholar]

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