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. 1983 Aug 25;11(16):5589–5602. doi: 10.1093/nar/11.16.5589

The binding of ribosomal protein S1 to S1-depleted 30S and 70S ribosomes. A fluorescence anisotropy study of the effects of Mg2+.

D J Goss, L J Parkhurst, A M Mehta, A J Wahba
PMCID: PMC326299  PMID: 6351011

Abstract

We have determined the equilibrium constants for the binding of AEDANS-labelled S1 to S1-depleted 30S and 70S ribosomes. For "tight" ribosomes, the association of S1 increases with the sixth power of Mg2+ concentration, but for 30S subunits and "loose" ribosomes, there is virtually no dependence of the association on Mg2+ over the same concentration range, 2-10 mM in Mg2+. The binding of S1 to 70S ribosomes at 10 mM Mg2+ is stabilized by 2 kcal/mol compared to the binding to 30S subunits. When intact S1 binds to tight ribosomes, the fluorescence anisotrophy is that for virtually complete rotational immobilization. The anisotropies vary considerably with the preparation and treatment of both S1 and ribosomes and these variations are detailed here. The results suggest the linkage of Mg2+-dependent conformational changes in the intact ribosomes, perhaps including rRNA, and the binding of S1.

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Selected References

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  1. Bear D. G., Ng R., Van Derveer D., Johnson N. P., Thomas G., Schleich T., Noller H. F. Alteration of polynucleotide secondary structure by ribosomal protein S1. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1824–1828. doi: 10.1073/pnas.73.6.1824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boileau G., Sommer A., Traut R. R. Identification of proteins at the binding site for protein S1 in 70 S ribosomes and 30 S subunits by cross-linking with 2-iminothiolane. J Biol Chem. 1981 Aug 10;256(15):8222–8227. [PubMed] [Google Scholar]
  3. Chu Y. G., Cantor C. R., Sawchyn I., Cole P. E. Segmental flexibility of ribosomal protein S1 bound to ribosomes and Q beta-replicase. FEBS Lett. 1982 Aug 23;145(2):203–207. doi: 10.1016/0014-5793(82)80168-3. [DOI] [PubMed] [Google Scholar]
  4. Chu Y. G., Cantor C. R. Segmental flexibility in Escherichia coli ribosomal protein S1 as studied by fluorescence polarization. Nucleic Acids Res. 1979;6(6):2363–2379. doi: 10.1093/nar/6.6.2363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dahlberg A. E. Two forms of the 30 S ribosomal subunit of Escherichia coli. J Biol Chem. 1974 Dec 10;249(23):7673–7678. [PubMed] [Google Scholar]
  6. Debey P., Hui Bon Hoa G., Douzou P., Godefroy-Colburn T., Graffe M., Grunberg-Manago M. Ribosomal subunit interaction as studied by light scattering. Evidence of different classes of ribosome preparations. Biochemistry. 1975 Apr 22;14(8):1553–1559. doi: 10.1021/bi00679a001. [DOI] [PubMed] [Google Scholar]
  7. Draper D. E., von Hippel P. H. Interaction of Escherichia coli ribosomal protein S1 with ribosomes. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1040–1044. doi: 10.1073/pnas.76.3.1040. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giorginis S., Subramanian A. R. The major ribosome binding site of Escherichia coli ribosomal protein S1 is located in its N-terminal segment. J Mol Biol. 1980 Aug 25;141(4):393–408. doi: 10.1016/0022-2836(80)90253-3. [DOI] [PubMed] [Google Scholar]
  9. Goss D. J., Parkhurst L. J., Wahba A. J. Kinetic studies of the rates and mechanism of assembly of the protein synthesis initiation complex. Biophys J. 1980 Oct;32(1):283–293. doi: 10.1016/S0006-3495(80)84957-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goss D. J., Parkhurst L. J., Wahba A. J. Kinetic studies on the interaction of chain initiation factor 3 with 70 S Escherichia coli ribosomes and subunits. J Biol Chem. 1982 Sep 10;257(17):10119–10127. [PubMed] [Google Scholar]
  11. Görisch H., Goss D. J., Parkhurst L. J. Kinetics of ribosome dissociation and subunit association studied in a light-scattering stopped-flow apparatus. Biochemistry. 1976 Dec 28;15(26):5743–5753. doi: 10.1021/bi00671a010. [DOI] [PubMed] [Google Scholar]
  12. Hartley B. S. Strategy and tactics in protein chemistry. Biochem J. 1970 Oct;119(5):805–822. doi: 10.1042/bj1190805f. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hudson E. N., Weber G. Synthesis and characterization of two fluorescent sulfhydryl reagents. Biochemistry. 1973 Oct 9;12(21):4154–4161. doi: 10.1021/bi00745a019. [DOI] [PubMed] [Google Scholar]
  14. Kimura M., Foulaki K., Subramanian A. R., Wittmann-Liebold B. Primary structure of Escherichia coli ribosomal protein S1 and features of its functional domains. Eur J Biochem. 1982 Mar;123(1):37–53. doi: 10.1111/j.1432-1033.1982.tb06495.x. [DOI] [PubMed] [Google Scholar]
  15. Laughrea M., Moore P. B. On the relationship between the binding of ribosomal protein S1 to the 30 S subunit of Escherichia coli and 3' terminus of 16 S RNA. J Mol Biol. 1978 Jun 5;121(4):411–430. doi: 10.1016/0022-2836(78)90391-1. [DOI] [PubMed] [Google Scholar]
  16. Laughrea M., Moore P. B. Physical properties of ribosomal protein S1 and its interaction with the 30 S ribosomal subunit of Escherichia coli. J Mol Biol. 1977 May 25;112(3):399–421. doi: 10.1016/s0022-2836(77)80189-7. [DOI] [PubMed] [Google Scholar]
  17. Lipecky R., Kohlschein J., Gassen H. G. Complex formation between ribosomal protein S1, oligo-and polynucleotides: chain length dependence and base specificity. Nucleic Acids Res. 1977 Oct;4(10):3627–3642. doi: 10.1093/nar/4.10.3627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Michalski C. J., Sells B. H., Chowdhury R. M., Wahba A. J. Molecular morphology of ribosomes. Influence of chain-initiation factor IF-3 and ribosomal protein S1 on ribosome conformation. Eur J Biochem. 1978 Sep 1;89(2):589–594. doi: 10.1111/j.1432-1033.1978.tb12563.x. [DOI] [PubMed] [Google Scholar]
  19. Mülsch A., Colpan M., Wollny E., Gassen H. G., Riesner D. Mechanism of the interaction between ribosomal protein S1 and oligonucleotides. Nucleic Acids Res. 1981 May 25;9(10):2367–2385. doi: 10.1093/nar/9.10.2367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schnier J., Kimura M., Foulaki K., Subramanian A. R., Isono K., Wittmann-Liebold B. Primary structure of Escherichia coli ribosomal protein S1 and of its gene rpsA. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1008–1011. doi: 10.1073/pnas.79.4.1008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sobura J. E., Chowdhury M. R., Hawley D. A., Wahba A. J. Requirement of chain initiation factor 3 and ribosomal protein S1 in translation of synthetic and natural messenger RNA. Nucleic Acids Res. 1977 Jan;4(1):17–29. doi: 10.1093/nar/4.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Subramanian A. R., Rienhardt P., Kimura M., Suryanarayana T. Fragments of ribosomal protein S1 and its mutant form m1-S1. Localization of nucleic-acid-binding domain in the middle region of S1. Eur J Biochem. 1981 Oct;119(2):245–249. doi: 10.1111/j.1432-1033.1981.tb05600.x. [DOI] [PubMed] [Google Scholar]
  23. Suryanarayana T., Subramanian A. R. Functional domains of Escherichia coli Ribosomal Protein S1. Formation and characterization of a fragment with ribosome-binding properties. J Mol Biol. 1979 Jan 5;127(1):41–54. doi: 10.1016/0022-2836(79)90458-3. [DOI] [PubMed] [Google Scholar]
  24. Szer W., Hermoso J. M., Boublik M. Destabilization of the secondary structure of RNA by ribosomal protein S1 from Escherichia coli. Biochem Biophys Res Commun. 1976 Jun 7;70(3):957–964. doi: 10.1016/0006-291x(76)90685-9. [DOI] [PubMed] [Google Scholar]
  25. Szer W., Hermoso J. M., Leffler S. Ribosomal protein S1 and polypeptide chain initiation in bacteria. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2325–2329. doi: 10.1073/pnas.72.6.2325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Szer W., Leffler S. Interaction of Escherichia coli 30S ribosomal subunits with MS2 phage RNA in the absence of initiation factors. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3611–3615. doi: 10.1073/pnas.71.9.3611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tal M., Aviram M., Kanarek A., Weiss A. Polyuridylic acid binding and translating by Escherichia coli ribosomes: stimulation by protein I, inhibition by aurintricarboxylic acid. Biochim Biophys Acta. 1972 Oct 27;281(3):381–392. doi: 10.1016/0005-2787(72)90452-2. [DOI] [PubMed] [Google Scholar]
  28. Thomas J. O., Kolb A., Szer W. Structure of single-stranded nucleic acids in the presence of ribosomal protein S1. J Mol Biol. 1978 Aug 5;123(2):163–176. doi: 10.1016/0022-2836(78)90319-4. [DOI] [PubMed] [Google Scholar]
  29. Visentin L. P., Hasnain S., Gallin W. Ribosomal protein S1/S1A in bacteria. FEBS Lett. 1977 Jul 15;79(2):258–263. doi: 10.1016/0014-5793(77)80799-0. [DOI] [PubMed] [Google Scholar]
  30. Wahba A. J., Miller M. J. Chain initiation factors from Escherichia coli. Methods Enzymol. 1974;30:3–18. doi: 10.1016/0076-6879(74)30003-1. [DOI] [PubMed] [Google Scholar]
  31. Wahba A. J., Miller M. J., Niveleau A., Landers T. A., Carmichael G. G., Weber K., Hawley D. A., Slobin L. I. Subunit I of G beta replicase and 30 S ribosomal protein S1 of Escherichia coli. Evidence for the identity of the two proteins. J Biol Chem. 1974 May 25;249(10):3314–3316. [PubMed] [Google Scholar]
  32. 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]
  33. Yuan R. C., Steitz J. A., Moore P. B., Crothers D. M. The 3' terminus of 16S rRNA: secondary structure and interaction with ribosomal protein S1. Nucleic Acids Res. 1979 Dec 20;7(8):2399–2418. doi: 10.1093/nar/7.8.2399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. van Duin J., van Knippenberg P. H. Functional heterogeneity of the 30 S ribosomal subunit of Escherichia coli. 3. Requirement of protein S1 for translation. J Mol Biol. 1974 Mar 25;84(1):185–195. doi: 10.1016/0022-2836(74)90221-6. [DOI] [PubMed] [Google Scholar]

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